{"id":59226,"date":"2024-06-25T11:42:03","date_gmt":"2024-06-25T11:42:03","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=59226"},"modified":"2024-09-24T03:57:36","modified_gmt":"2024-09-24T03:57:36","slug":"anti-inflammatory-and-antioxidant-properties-of-the-mucuna-sanjappae-seeds-in-the-rat-model-and-in-vitro-assays","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol17no2\/anti-inflammatory-and-antioxidant-properties-of-the-mucuna-sanjappae-seeds-in-the-rat-model-and-in-vitro-assays\/","title":{"rendered":"Anti-Inflammatory and Antioxidant Properties of the Mucuna sanjappae Seeds in the Rat Model and In Vitro Assays"},"content":{"rendered":"\n

Introduction<\/strong><\/p>\n\n\n\n

Inflammation can be defined as essential\nresponse exhibited by host after tissue injury or infection. However, its longstanding\npersistence may result in chronic diseases like cancer, cardiovascular\ndiseases, neurological disorders, diabetes, pulmonary diseases, and arthritis1-7<\/sup>.\nInflammation is characterized by pro-inflammatory enzymes, chemokines, cytokines,\nand certain signal proteins generated in response to infection or injury. Management\nof inflammatory diseases today represents an important medical problem since\ncurrently used non-steroidal anti-inflammatory drugs (NSAIDs) has several other\nadverse effects commonly known as gastroenteropathy1,7<\/sup>. Hence, identification\nof novel and effective therapies for safer management of inflammatory diseases\nremains an urgent priority. Natural plant wealth is continually being\ninvestigated for novel bioactive molecules with therapeutic properties. In\ncontrast to modern synthetic drugs, natural bioactive are cost effective and provides\nsignificant protection from diseases without secondary adverse complications. Therefore,\nresearchers are investigating plant-based drugs which can provide promising anti-inflammatory\nactivity with lower secondary complications8<\/sup>. Numerous reports\nendorse the use of various plants for treating inflammation1,9<\/sup>.           <\/p>\n\n\n\n

Genus Mucuna<\/em> belongs to family fabaceae, popularly known as cowitch,\nkapikachu and atmagupta. Mucuna <\/em>species\nare recognized for their itching properties due to hairs present on the pods. Since\nancient time, Indian ayurveda system of medicine uses seeds of Mucuna <\/em>for the management of different\ntypes of diseases and disorders. Remarkable research has been carried out on Mucuna pruriens <\/em>particularly for its anti-Parkinson\u2019s,\nanti-infertility, anti-venom, anti-inflammatory and anti-bacterial activity7,10-16<\/sup>.\nMucuna <\/em>seed powder is very common ingredient in various ayurvedic\nformulations marked in India and across the world. However, exploitation of M.\npruriens <\/em>at large scale may affect ecosystem adversely and due to limited\navailability, final product cost may also increase. To avoid this, there is\nincreasing interest in investigating hidden potential of other underutilized Mucuna <\/em>spp. as a promising alternative\ntherapeutic agent. Previously, our research group have successfully reported Mucuna\n<\/em>species including M. macrocarpa, M. bracteata<\/em>, M. imbricata <\/em>and\nM. sanjappae<\/em> etc. for their nutritional and medicinal benefits15-24<\/sup>.\nIn 2019, we have reported L-DOPA (L-3,4-dihydroxyphenylalanine), an FDA\napproved anti-Parkinson\u2019s drug in seeds of different Mucuna <\/em>species found in Indian contingent and proved that number of\nMucuna <\/em>species possesses higher level\nof L-DOPA as compared to the commonly used M.\npruriens<\/em>25<\/sup>. <\/p>\n\n\n\n

M.\nsanjappae<\/em> is endemic plant species found in Western\nGhats region of Maharashtra, India which belongs to genus Mucuna<\/em>26<\/sup>.\nIn previous studies we have demonstrated that M. sanjappae <\/em>seeds possess promising level of nutritional\ncomponents with gross energy 383 kcal and around 5.43 g of protein. Moreover,\nit contains about 7.3 % L-DOPA and other important primary and secondary\nmetabolites, minerals, important phenolics etc. Furthermore, anti-Parkinson\u2019s\nactivity of M. sanjappae <\/em>seed extract\nin Parkinson\u2019s disease (PD) mice model intoxicated by MPTP is reported18,19<\/sup>.\nM. sanjappae <\/em>seed extract could successfully ameliorate PD symptoms\ndeveloped by MPTP toxicity. However, till date, there is no data available on\nthe effect of M. sanjappae <\/em>on\ninflammatory diseases and oxidative stress using in vitro <\/em>or in vivo <\/em>model.\nCarrageenan induced rat paw edema model is popular method of anti-inflammatory\nstudies of natural as well as synthetic drugs1<\/sup>. Hence, present\nefforts have been made to examine anti-inflammatory and antioxidant properties of\nM. sanjappae <\/em>seed on carrageenan\ninduced rat paw edema model. <\/p>\n\n\n\n

Material and Methods<\/strong><\/p>\n\n\n\n

Chemicals\nand reagents<\/strong><\/p>\n\n\n\n

Analytical grade solvents and chemicals\nwere used the study. Carrageenan and enzyme-linked immunosorbent assay (ELISA)\ncytokine kits for TNF-\u03b1 and IL-10 measurement were obtained from Sigma-Aldrich,\nUSA. Diclofenac (Standard anti-inflammatory drug) was obtained from Recon,\nBangalore, India respectively. <\/p>\n\n\n\n

Plant\nmaterial and preparation of drugs for administration<\/strong><\/p>\n\n\n\n

The pods of M. sanjappae<\/em> were collected from its original location (Pune\ndistrict, Western Ghats, Maharashtra, India). The herbarium was carefully\nprepared and stored at the herbarium center of the Botany department, Shivaji\nUniversity, Kolhapur under the guidance of taxonomist Prof S. R. Yadav. <\/p>\n\n\n\n

After removing the healthy seeds from\nthe pods, fine powder was prepared. Extract\nof seed was produced by adding and macerating 1g M. sanjappae<\/em>\nseed powder in 100 ml D\/W in the mortar and\npestle. Further, sonication for 15min and centrifugation\nat 10000 rpm for 10 min was carried out. Supernatant was carefully separated\nand stored for further use. Effective yield was calculated by evaporating\nwater. Importantly, seed extract and standard drug diclofenac were prepared\nfreshly at the time of dosing. <\/p>\n\n\n\n

In vitro<\/em><\/strong> anti-inflammatory\nactivity<\/strong><\/p>\n\n\n\n

Bovine\nserum albumin (BSA) anti-denaturation assay<\/strong><\/p>\n\n\n\n

This test was performed using a\nmethod defined earlier with slight modifications27<\/sup>. Various concentrations\nof seed extract and standard drug diclofenac were reacted with 1ml of 1% BSA\nsolution prepared in 50mM Tris buffer (pH 6.5). Incubation was carried out at\n37\u00b0C for 20 min and further heating at 64\u00b0C in water bath till mixture get\nturbid (around 5 to 10 min). Finally, tubes were cooled, and absorbance of\ngenerated turbidity was measured at 660 nm. D\/W was used as a control. Following\nformula was used to calculate denaturation inhibition percentage:    <\/p>\n\n\n\n

% denaturation inhibition = A\n(control) \u2013 A (sample) \/ A (control) X 100<\/p>\n\n\n\n

Where, A (control): Absorbance of the\ncontrol; A (sample): Absorbance of samples.<\/p>\n\n\n\n

Hypotonicity-induced HRBC\nmembrane stabilization method<\/strong><\/p>\n\n\n\n

Capacity of M. sanjappae <\/em>seed\nto protect hypotonicity encouraged human red blood cell membrane protection have\nbeen determined28<\/sup>. Various concentrations of M. sanjappae <\/em>seed\nextract were made into 1 mL using distilled water in a tube. Initially, 0.5ml\nof 10% HRBC suspension and 0.5 mL of 0.25% hyposaline were added to each tube.\nThen, mixture was incubated at static condition for 30 mins at 37\u00b0C and\ncentrifuged at 3000 rpm for 20 mins at 4\u00b0C. The amount of hemoglobin in the\nsupernatant was performed at a wavelength of 560 nm. Working solution of\nstandard drug aspirin was prepared in 0.2 M phosphate buffer (1 mL) at various\nconcentration ranging from 100 \u00b5g to 500 \u00b5g. To induce complete hemolysis (100%)\nwithout any sample or drug, a control was prepared using distilled water as a\nreplacement for hyposaline. To calculate the percentage of HRBC hemolysis and\nevaluate the degree of membrane stabilization or protection, below given formula\nwas employed:<\/p>\n\n\n\n

% of hemolysis = Absorbance of test\/\nAbsorbance of control x 100<\/p>\n\n\n\n

% protection = 100 \u2013 (% of\nhemolysis)  <\/p>\n\n\n\n

In vivo <\/em><\/strong>anti-inflammatory\nactivity<\/strong><\/p>\n\n\n\n

Rats were divided into seven groups (6\nrats per polypropylene cage) housed under controlled conditions (Relative humidity\n44\u201356 %, temperature of 25\u00b12 \u00b0C, and 12 h light\/ dark cycles). Standard diet\nand water ad libitum <\/em>was provided to the experimental animals. The experiment\nwas started after proper acclimatization of animals in the laboratory\nenvironment after a week. Details of randomization, grouping and drug dosing are\ngiven in table 1. Seven days prior dosing was carried out by M. sanjappae <\/em>seedextractin group IV, V, VI and VII. On the 8th<\/sup> day rats\nwere kept fasted but water was provided ad\nlibitum. <\/em>Extract was administered 2hr before inducing the inflammation by\ncarrageenan (0.9%) through sub plantar way and further investigation was\naccomplished.<\/p>\n\n\n\n

Table 1: Randomization and grouping of animals for in vivo <\/em>study<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Group number <\/strong><\/p>\n<\/td>\n

\n

Group name <\/strong><\/p>\n<\/td>\n

\n

Dose content <\/strong><\/p>\n<\/td>\n

\n

Number of animals<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

           I<\/p>\n<\/td>\n

\n

Normal control<\/p>\n<\/td>\n

\n

Vehicle solvent (D\/W)<\/p>\n<\/td>\n

\n

6<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/p>\n<\/td>\n

\n

Carrageenan control<\/p>\n<\/td>\n

\n

100 \u00b5l (0.9% carrageenan) # <\/sup><\/p>\n<\/td>\n

\n

6<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/p>\n<\/td>\n

\n

Diclofenac<\/p>\n<\/td>\n

\n

10mg\/kg BW<\/p>\n<\/td>\n

\n

6<\/p>\n<\/td>\n<\/tr>\n

\n

IV<\/p>\n<\/td>\n

\n

Test Dose-1 50mg\/kg BW*<\/p>\n<\/td>\n

\n

50mg\/kg BW<\/p>\n<\/td>\n

\n

6<\/p>\n<\/td>\n<\/tr>\n

\n

V<\/p>\n<\/td>\n

\n

Test Dose-2 100mg\/kg BW<\/p>\n<\/td>\n

\n

100mg\/kg BW<\/p>\n<\/td>\n

\n

6<\/p>\n<\/td>\n<\/tr>\n

\n

VI<\/p>\n<\/td>\n

\n

Test Dose-3 200mg\/kg BW<\/p>\n<\/td>\n

\n

200mg\/kg BW<\/p>\n<\/td>\n

\n

6<\/p>\n<\/td>\n<\/tr>\n

\n

VII<\/p>\n<\/td>\n

\n

Sham Control<\/p>\n<\/td>\n

\n

200mg\/kg BW<\/p>\n<\/td>\n

\n

6<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

BW* – Body Weight of animals.<\/p>\n

# – 0.9% carrageenan were prepared in saline solution<\/p>\n\n\n

Rat paw edema measurement <\/strong><\/p>\n\n\n\n

The inflammation in terms of swelling\nof carrageenan induced foot of animal was measured using plethysmometer (UGO\nBasile, Italy) as a water displacement in ml. The measurement was done at 0, 2,\n4 and 6 hr. The decrease in paw volume was compared to the vehicle control. The\npercentage of inhibition in seed water extract and diclofenac treated group was\ncompared with carrageenan induced group. <\/p>\n\n\n\n

Inhibition Percentage (%) =\n[Carrageenan treated group-test drug group \/ Carrageenan treated group] X 100<\/p>\n\n\n\n

Inflammatory\nbiomarkers and Oxygen radical absorbance capacity (ORAC) assay  <\/strong><\/p>\n\n\n\n

 TNF-\u03b1 and IL-10 concentration in serum of\ncontrol and drugs treated respective groups was determined using ELISA kit. The\nexperiment was performed as per instructions of manufacturer. TNF-\u03b1 and IL-10 level\nwas expressed as picogram per milligram (pg\/mg). <\/p>\n\n\n\n

Antioxidant level of serum samples in\ntreated and control animal group was carried out by ORAC method29<\/sup>. Shortly,\n25 \u03bcl of serum sample added in fresh 150 \u03bcl of 10nM fluorescein solution and allowed\nto stand at 37 0<\/sup>C for 30 min. After incubation, 25 ml of AAPH\nsubstrate (500mM) mixed and the fluorescence was calculated for 150 min at 485\nand 520nm (Excitation and emission wavelengths respectively) using microplate\nreader. Standard trolox was used as a and results were expressed as micromoles\nof Trolox equivalents (TE) per liter of sample.<\/p>\n\n\n\n

Phytochemical\nanalysis<\/strong><\/p>\n\n\n\n

The total polyphenol level of M. sanjappae<\/em> seeds was determined\nspectrophotometrically30<\/sup> and represented as mg of gallic acid\nequivalent per gram (mg GAE g\u20131<\/sup>) of dry mass. Theflavonoids content was analyzed31<\/sup>\nand results represented as milligram of quercetin equivalents per gram (mg QUE\ng-1<\/sup>) of dry weight. Proanthocyanidin examined and reported as\ncatechin equivalents per gram (mg CAE g-1<\/sup>) of dry weight32<\/sup>.\nThe phytic acid was determined and absorbance was measured at 500 nm33<\/sup>.\nTannin and saponin level also studied according to methods reported earlier34,35<\/sup>.<\/p>\n\n\n\n

Statistical\nAnalysis <\/strong><\/p>\n\n\n\n

GraphPad Prism 5 is used for data\nanalysis. All the results were represented Mean \u00b1 SEM. P-values of less than\n0.05 were considered as significant.<\/p>\n\n\n\n

Results and Discussion <\/strong><\/p>\n\n\n\n

Nature has gifted us with medicinally\nimportant, inexhaustible sources of secondary metabolites, including alkaloids,\nterpenoids, phenolics, saponins, and other classes of organic compounds. These\nphytometabolites have tremendous health benefits for the overall growth and the\nmanagement of diseases. Over time, experimental procedures and tools for the\nisolation, characterization, validation, and development of drugs for disease\nhave been well established36<\/sup>. The present attempt was aimed to find\nout the anti-inflammatory and antioxidant properties of M. sanjappae<\/em>\nseeds for future natural drug development.     <\/p>\n\n\n\n

In vitro anti-inflammatory activity<\/strong><\/p>\n\n\n\n

Preliminary screening of anti-inflammatory potential was performed using\ntable assay as given below:<\/p>\n\n\n\n

Bovine serum albumin (BSA) anti-denaturation\npotential:<\/strong><\/p>\n\n\n\n

Denaturation of proteins causes several inflammatory responses in the body. Many synthetic drug molecules show promising anti-inflammatory activity37<\/sup> but are known to cause secondary complications after long-term use38<\/sup>. Hence, the use of plant-mediated drugs may prove more advantageous than their synthetic counterparts. M. sanjappae<\/em> seed extract has shown inhibition of heat-induced albumin denaturation activity at different concentrations, as shown in Fig. 1. M. sanjappae<\/em> extracts showed strong inhibition of albumin denaturation (87.73\u00b13.81%) at 500\u03bcg\/ml concentration. The standard drug diclofenac showed 94.82\u00b1 1.79% inhibition at 500 \u03bcg concentration. The results suggested good anti-inflammatory activity of M. sanjappae<\/em> seed extract.<\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 1: Heat induced BSA anti-denaturation activity M. sanjappae <\/em>seed water extract in % comparison with standard drug Diclofenac (Mean \u00b1  D, n=3).<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

HRBC Membrane stability potential<\/strong><\/p>\n\n\n\n

Stabilisation of lysosomal membrane is an essential\nprocess in the regulation of inflammation process. It occurs by preventing the release\nof lysosomal components of active neutrophils including bactericidal enzymes\nand proteases. Red blood cell membrane bears resemblance to lysosomal membrane28<\/sup>,\ntherefore understanding the stability of RBC membrane by our drug of interest\ngives idea about its potential to protect lysosomal membrane to prevent release\nof inflammatory response markers. Haemoglobin released in the supernatant in\nthe control tube (without std drug or plant extract) due to bursting of red\nblood cells, while the yellow supernatant in the plant extract tube indicates\nthe stabilisation of HRBCs. Our results demonstrated that M.\nsanjappae <\/em>seed extract could protect the HRBC membrane with maximum\nprotection at 500ug M. sanjappae <\/em>seed\nextract (60.47\u00b12.1 %) as depicted in Fig. 2. As\nconcentration of M. sanjappae<\/em> seed extract was increased, HRBC membrane\nstability was also increased suggesting concentration dependent stabilization\nactivity by M. sanjappae<\/em> seed. Standard drug aspirin showed superior\nHRBC membrane protection capacity as compared to M.\nsanjappae <\/em>seed extract. The maximum protection by aspirin at 500ug concentration was 99.56\u00b11.01\nand it also represented dose dependent increase in activity.<\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 2: HRBC membrane stability assay of different concentration of M. sanjappae <\/em>seed <\/i>water extract in comparison with standard drug Aspirin (Mean \u00b1  D, n=3)<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

In vivo <\/em><\/strong>anti-inflammatory\nactivity<\/strong><\/p>\n\n\n\n

Carrageenan is a sulfonated\npolysaccharide widely used in food industries. It is extracted from red seaweed\nalgae. It does not have any nutritional potential but is preferentially used as\nthickening, gelling and emulsifying agent39<\/sup>. Several studies have\nsuccessfully reported use of carrageenan as an inflammatory agent to induce\nacute paw edema1<\/sup>. It is a simple and effective mean of assessing\nanti-inflammatory properties of drug. <\/p>\n\n\n\n

Paw\nedema measurement<\/strong><\/p>\n\n\n\n

Determination of paw edema level\nafter carrageenan toxicity and its furth treatment using oral dose of M.\nsanjappae <\/em>seed extract was evaluated. Carrageenan generates acute paw edema\nwhich can be seen as redness and swelling at the site of injection. In the\npresent study, paw edema was induced by using carrageenan and measured at 0hr,\n2hr, 4hr and 6hr post injection. Carrageenan toxicity successfully developed\ninflammation in the rat paw which was evident from redness and swelling at the\nsite of induction with highest paw thickness at 4hr (9.44\u00b10.25). Vehicle\ncontrol animal group does not show edema. Animal group treated with different\ndoses of M. sanjappae <\/em>seed extract exhibited significant reduction in\nthe paw edema with maximum result at 200mg\/kg body weight (Table 2). It showed\n53.49% of edema inhibition after 4 hr treatment. Standard drug showed maximum\nedema inhibition (54.94%) at 6hr.The anti-inflammatory potential of M.\nsanjappae <\/em>seed extract was observed very close to the standard drug Diclofenac control group. Dose\ndependent increase in the anti-inflammatory activity of M. sanjappae <\/em>seed\nextract in respect to edema reduction was clear from the study. Sham control group\nwas given the highest dose of M. sanjappae <\/em>seed extract (200mg\/kg body weight) and does not show\nany inflammatory symptoms during the experiments. Our\nfinding supports anti-inflammatory potential of M. sanjappae <\/em>seed and its traditional use in the management of\ninflammation related disorders. <\/p>\n\n\n\n

 Table 2: Effect of M. sanjappae<\/em> on carrageenan induced rat paw edema.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
 <\/td>\n\n

 0hr<\/strong><\/p>\n<\/td>\n

\n

2hr<\/strong><\/p>\n<\/td>\n

\n

4hr<\/strong><\/p>\n<\/td>\n

\n

6hr<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Group I: Vehicle control<\/p>\n<\/td>\n

\n

4.11\u00b10.04<\/p>\n<\/td>\n

\n

4.16\u00b10.05<\/p>\n<\/td>\n

\n

3.97\u00b10.06<\/p>\n<\/td>\n

\n

4.05\u00b10.04<\/p>\n<\/td>\n<\/tr>\n

\n

Group II: Carrageenan Control<\/p>\n<\/td>\n

\n

4.41\u00b10.08<\/p>\n<\/td>\n

\n

8.23\u00b10.07<\/p>\n<\/td>\n

\n

9.44\u00b10.25<\/p>\n<\/td>\n

\n

8.9\u00b10.39<\/p>\n<\/td>\n<\/tr>\n

\n

Group III: Diclofenac control<\/p>\n<\/td>\n

\n

3.89\u00b10.04<\/p>\n<\/td>\n

\n

4.51\u00b10.06***  <\/sup>(47.0)<\/p>\n<\/td>\n

\n

4.26\u00b10.21*** <\/sup>(54.87)<\/p>\n<\/td>\n

\n

4.01\u00b10.18***<\/sup> (54.94)<\/p>\n<\/td>\n<\/tr>\n

\n

Group IV: 50mg test drug$<\/sup><\/p>\n<\/td>\n

\n

3.91\u00b10.14<\/p>\n<\/td>\n

\n

6.02\u00b10.36***<\/sup> (26.85)<\/p>\n<\/td>\n

\n

5.24\u00b10.19***<\/sup> (44.49)<\/p>\n<\/td>\n

\n

5.04\u00b10.2***<\/sup> (43.37)<\/p>\n<\/td>\n<\/tr>\n

\n

Group V: 100mg test drug<\/p>\n<\/td>\n

\n

4.28\u00b10.09<\/p>\n<\/td>\n

\n

5.99\u00b10.25***<\/sup> (27.21)<\/p>\n<\/td>\n

\n

5.27\u00b10.33*** <\/sup> (44.17)<\/p>\n<\/td>\n

\n

5.11\u00b10.29*** <\/sup>(42.58)<\/p>\n<\/td>\n<\/tr>\n

\n

Group VI: 200mg test drug<\/p>\n<\/td>\n

\n

3.81\u00b10.02<\/p>\n<\/td>\n

\n

4.53\u00b10.15***<\/sup> (44.95)<\/p>\n<\/td>\n

\n

4.39\u00b10.22***<\/sup> (53.49)<\/p>\n<\/td>\n

\n

4.18\u00b10.31***<\/sup> (53.03)<\/p>\n<\/td>\n<\/tr>\n

\n

Group VII: SHAM control<\/p>\n<\/td>\n

\n

4.49\u00b10.05<\/p>\n<\/td>\n

\n

4.47\u00b10.08<\/p>\n<\/td>\n

\n

4.52\u00b10.1<\/p>\n<\/td>\n

\n

4.36\u00b10.11<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

$: M. sanjappae <\/em>seed water extract. Values have been represented as mean\u00b1SEM, n=6 in each group. * p<0.05; ** p<0.01; *** p<0.001 when compared with carrageenan induced control. Values shown in parentheses represent the percent (%) reduction in paw edema in comparison with carrageenan induced control.<\/p>\n\n\n

Effect of M. sanjappae <\/em>treatment on pro-inflammatory cytokine TNF-\u03b1 <\/strong><\/p>\n\n\n\n

TNF-\u03b1 is a mediator of inflammatory\nresponses playing a key role in the development of innate immune system via\nactivating macrophages, T cells and secretion of other inflammatory cytokines.\nIn the carrageenan induced acute inflammatory model, complement system\nstimulation along with inflammatory mediators\u2019 synthesis are major events 40<\/sup>.\nSerum TNF-\u03b1 level was determined after 6hr post carrageenan treatment (Fig 3a).\nThere was considerable increase in TNF-\u03b1 (89\u00b18.13 pg)\nin serum due to inflammatory response after carrageenan injection confirming\nits pro-inflammatory properties (Group II). However, treatment of M.\nsanjappae <\/em>seed extract at various doses significantly decreased TNF-\u03b1 level.\nAmong tested doses, 100 and 200mg dose significantly reduced (p<0.01) TNF-\u03b1\nlevel (59.5\u00b14.48\npg and 57\u00b11.96\npg respectively) in animal (Fig 3a) and we found superior results as compared to\nstandard drug diclofenac (61.3\u00b16.61 pg). Vehicle control and Sham\ncontrol represented normal TNF- \u03b1 level. According to the results, it can be concluded\nthat, M. sanjappae <\/em>has potential of\nreducing the inflammation caused by external toxic compounds and possesses\npotential to use for managing inflammatory diseases. <\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 3a: Effect of M. sanjappae <\/em>seed extract on TNF- \u03b1 level in serum of carrageenan induced rat at 6hr.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Values\nare represented as Mean\u00b1SEM (n=6). **p<0.01, **p<0.01,\n***p<0.001 when compared with carrageenan induced group.  ##<\/sup>p<0.01, ###<\/sup><0.001\nwhen compared with vehicle control group (X axis indicates group number which\nare shown in detail in Table no. 1)<\/p>\n\n\n\n

Effect\nof M. sanjappae <\/em>treatment on anti-inflammatory\ncytokine IL-10 <\/strong><\/p>\n\n\n\n

IL-10 is an important\nanti-inflammatory cytokine which attenuates the activity of pro-inflammatory\nmarkers including TNF- \u03b141<\/sup>. In the present study, in contrast to\nTNF- \u03b1, IL-10 level was increased in Diclofenac and M. sanjappae <\/em>seed\nextract treated animal groups (Fig. 3b). Reduction in IL-10 in carrageenan\ninduced group (II), suggests, carrageenan exerts its anti-inflammatory response\nthrough suppression of anti-inflammatory cytokines. Positive effect of M.\nsanjappae <\/em>seed extract was validated from considerable augmented IL-10 level\n(p<0.001) with maximum activity at 100 and 200mg M. sanjappae <\/em>seed\nextract dose. Vehicle control (I) and SHAM control (VII) exhibited normal level\nof IL-10 which was higher than carrageenan induced group II. Overall, our study\nsignifies M. sanjappae <\/em>exerts its\nanti-inflammatory action by suppressing pro-inflammatory cytokines and\nexpressing anti-inflammatory cytokines in the serum of carrageenan toxicated\nrat. <\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 3b: Effect of M. sanjappae <\/em>seed extract on IL-10 level in serum of carrageenan induced rat at 6hr.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Results\nare represented as Mean\u00b1SEM (n=6). ***p<0.001 when\ncompared with carrageenan induced group. ###<\/sup>p<0.001 in comparison\nwith vehicle control group. (X axis indicates group\nnumber which are shown in detail in Table no. 1).<\/p>\n\n\n\n

Oxygen radical absorbance capability (ORAC) assay<\/strong><\/p>\n\n\n\n

Serum antioxidant levels of different\nanimal groups is studied by oxygen radical absorbance capacity method1<\/sup>.\nSerum antioxidant activity was significantly (p<0.001) elevated in M.\nsanjappae <\/em>seed extract treated animal group. The higher activity was found\nfor 100 and 200mg\/kg B\/W dose. There was also significant increase in serum\nantioxidant capacity in sham control animal suggesting M. sanjappae <\/em>seed\npossesses secondary metabolites which are enhancing antioxidant properties in\nthe animal. Carrageenan control showed lower level of serum antioxidant\nactivity (468\u00b131.6\u00b5<\/a>M Trolox equiv\/L) suggesting\noxidative stress is get generated due to toxicity of carrageenan. Different\ndoses of M. sanjappae <\/em>seed extract showed 560\u00b122.4, 905\u00b153.8 and\n1105\u00b142.4 \u00b5M Trolox equiv\/L respectively showing concentration dependent\nincrease in oxygen radical absorbance potential (Fig 4).  <\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 4: Effect of M. sanjappae <\/em>seed extract on serum oxygen radical absorbance capacity in carrageenan induced rat at 6hr.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Results\nare represented as Mean\u00b1SEM (n=6). ***p<0.001 when\ncompared with carrageenan induced group. \n###<\/sup>p<0.001 when compared with vehicle control group     <\/p>\n\n\n\n

Human disorders show pathogenesis\nthrough cellular inflammation and oxidative stress mediated cell degeneration. An\ninbuilt antioxidant system comprising antioxidant molecules and enzymes performs\na crucial role in removing free radicals7,42<\/sup>. But increased\noxidative stress because of environmental toxins, genetic changes, or unknown\ncauses results in cell components\/organelle to degenerate or alter leading to\napoptosis of cell. In such cases, supplementary antioxidants through food or\ndrugs become essential way of disease management. Present study demonstrated M. sanjappae <\/em>seeds contains vital phyto-metabolites\nwhich have capacity to induce anti-inflammatory and antioxidant properties required\nin the disease treatment. <\/p>\n\n\n\n

Phytochemical\nanalysis <\/strong><\/p>\n\n\n\n

Phenolics and flavonoids are\nconsidered as major secondary metabolites with prominent role as antioxidant\nand anti-inflammatory agents20,21,22,24<\/sup>. These secondary metabolites\nare with diverse biological activities combating diseases through multiple\npathways. Their role in managing diseases including neurodegenerative diseases is\nvia unstable superoxide radicals scavenging and cellular inflammation decreasing\npathway43<\/sup>. M. sanjappae<\/em>\nshowed 80.78\u00b12.56 mg GAE g-1<\/sup> of phenolics and 419.5\u00b17.18 mg QAE g-1<\/sup>\nof flavonoids. Earlier we have analyzed gallic acid, tannic acid, p-hydroxybenzoic\nacid and p-coumaric acid as major phenolic compounds present in M. sanjappae<\/em>\nseeds using HPLC20<\/sup>. M. sanjappae<\/em> seed possesses higher levels\nof flavonoids than polyphenols which may be attributed to the specificity, and\naccuracy of reaction and respective standard used for the reaction. Proanthocyanidin\nlevel was 2.14 \u00b1 0.13 mg CAE g-1<\/sup>. Proanthocyanidin is flavan-3-ol group\nhaving compound with strong antioxidant, anti-inflammatory, antihypertensive,\nantimicrobial and antiallergic activity44-46<\/sup>. M. sanjappae<\/em> seed\ncontains a higher concentration of phytic acid 197.23\u00b10.11 mg g-1<\/sup>. Phytic\nacid is metal chelating and anti-inflammatory in nature 47,48<\/sup>. Phytic\nacid decreased inflammation by reducing the level of NF-\u03baB and p-ERK in MPTP\nintoxicated PD mice model48<\/sup>. It is also natural iron chelating agent\nwhich prevents iron induced dopaminergic neuron degeneration in Parkinson\u2019s\ndisease49<\/sup>. M. sanjappae<\/em> beans showed 0.52\u00b10.11 mg g-1 <\/sup>and\n18.71 \u00b1 0.13 mg g-1 <\/sup>of tannin and saponin content respectively. Both\nof these compounds possess anti-inflammatory and antioxidant activity50,\n51<\/sup>.<\/p>\n\n\n\n

Investigation of traditional ayurveda\nknowledge with respect to disease management has lain to find novel drug molecules\nand related molecular pathways responsible for it. In modern medicine, single\nsynthetic drug is preferred for targeted and quick action. But on the other\nside, those synthetic molecules usually have several side effects to the\npatients. In this connection, plant-based drug therapy which comprises several\neffective drug molecules proves to be most effective and moreover exerts\nminimum side effects. Based on the present results, M. sanjappae<\/em> may prove as a promising lead for treating\ninflammatory diseases.<\/p>\n\n\n\n

Conclusion<\/strong><\/p>\n\n\n\n

Natural drugs isolated from plants show promising anti-inflammatory properties with little to no side effects. Local indigenous peoples have been using Mucuna<\/em> species as a staple food and medicinal purposes, mainly for Parkinson’s disease, male infertility, and snake bite treatment. However, there has been no elaborative investigation of the anti-inflammatory properties of M. sanjappae <\/em>using in vivo <\/em>or in vitro <\/em>model. The present study revealed the anti-inflammatory potential of M. sanjappae<\/em> seed extract by inhibiting pro-inflammatory cytokines and upregulating anti-inflammatory cytokines. The carrageenan-induced edema was reduced after treatment with M. sanjappae<\/em> seeds’ water extract. Furthermore, the antioxidant level was elevated after the treatment by M. sanjappae <\/em>extract. Phytochemical analysis confirmed presence of active secondary metabolites such as phenolics, flavonoids, phytic acid, saponins etc. Thus, the study strongly supported therapeutic potential of M. sanjappae<\/em> and suggests further molecular-level investigations for its future exploration as a pharmacological agent in the management of inflammation and oxidative stress-related diseases.<\/p>\n\n\n\n

Acknowledgements<\/strong><\/p>\n\n\n\n

We acknowledge Interactive Research School for Health Affairs (IRSHA), Bharati Vidyapeeth, Pune for providing animal house facility for the research work <\/p>\n\n\n\n

Conflict of Interest <\/strong><\/p>\n\n\n\n

Authors declare no conflict of interest.<\/p>\n\n\n\n

Funding source<\/strong><\/p>\n\n\n\n

This work was supported by Department\nof Biotechnology, Govt. of India for funding through DBT-IPLS program (Ref.\nNo.: BT\/PR4572\/INF\/22\/147). Mr. Ravishankar Patil thanks SERB, DST for\nproviding financial support through N-PDF (PDF\/2016\/002075). Mr. Chetan Aware acknowledges\nSUK-DBT IPLS program for the fellowship. Mr. Govind Vyavahare acknowledges\nShivaji University for DRS. Dr. Ruchika Kaul-Ghanekar would like to acknowledge\nDirector, IRSHA and ministry of AYUSH for providing financial support for the\nstudy. Prof. Vishwas Bapat is thankful to Indian National Science Academy, New\nDelhi, India for senior scientist fellowship.<\/p>\n\n\n\n

References<\/strong><\/p>\n\n\n\n

  1. Choudhari A, Raina P, Deshpande M, Wali A, Zanwar A, Bodhankar S, Kaul-Ghanekar R. Evaluating the anti-inflammatory potential of Tectaria cicutaria<\/em> L. rhizome extract in vitro<\/em> as well as in vivo.<\/em> Journal of Ethnopharmacology<\/em> 2013; 150, 215\u2013222. https:\/\/doi.org\/10.1016\/ j.jep.2013.08.025
    CrossRef<\/a><\/li>
  2. Liu X, Yin L, Shen s, Hou Y. Inflammation and cancer: paradoxical roles in tumorigenesis and implications in immunotherapies. Genes & Diseases<\/em> 2023; 10, 151-164.
     CrossRef <\/a><\/li>
  3. Alfaddagh A, Martin SS, Leucker TM, Michos ED, Blaha MJ, Lowenstein CJ, Jones SR, Toth PP. Inflammation and cardiovascular disease: From mechanisms to therapeutics. American Journal of Preventive Cardiology<\/em> 2020; 21; 4:100130. doi: 10.1016\/j.ajpc.2020.100130.
    CrossRef <\/a><\/li>
  4. Marriott E, Singanayagam A, El-Awaisi J. Inflammation as the nexus: exploring the link between acute myocardial infarction and chronic obstructive pulmonary disease. Frontiers in Cardiovascular Medicine<\/em> 2024; 11:2024 | https:\/\/doi.org\/10.3389\/fcvm.2024.1362564
     CrossRef <\/a><\/li>
  5. Bindu S, Mazumder S, Bandyopadhyay U. Non-steroidal anti-inflammatory drugs (NSAIDs) and organ damage: A current perspective. Biochemical<\/em> Pharmacology<\/em> 2020; 180:114147. doi: 10.1016\/j.bcp.2020.114147
    CrossRef <\/a><\/li>
  6. van den Bosch MHJ, Blom AB, van der Kraan PM. Inflammation in osteoarthritis: Our view on its presence and involvement in disease development over the years. Osteoarthritis Cartilage<\/em> 2024; 32:355-364. doi: 10.1016\/j.joca.2023.12.005
     CrossRef <\/a><\/li>
  7. Rai SN, Birla H, Singh S, Zahra W, Patil R, Jadhav J, Rao GM, SinghSP. Mucuna pruriens<\/em> protects against MPTP intoxicated neuroinflammation in Parkinson\u2019s disease through NF-\u03baB \/pAKT signaling pathways. Frontiers in Aging Neuroscience <\/em>2017;19 (9) https:\/\/doi.org\/10.3389\/fnagi.2017.00421
     CrossRef <\/a><\/li>
  8. Nunes CdR, Barreto Arantes M, Menezes de Faria Pereira S, Leandro da Cruz L, de Souza Passos M, Pereira de Moraes L, Vieira IJC, Barros de Oliveira D. Plants as Sources of Anti-Inflammatory Agents. Molecules. 2020; 25(16):3726. https:\/\/doi.org\/10.3390\/ molecules25163726
     CrossRef <\/a><\/li>
  9. Zhen J, Guo Y, Villani T, Carr S, Brendler, Mumbengegwi D, Kong AT, Simon J, Wu W. Phytochemical Analysis and Anti-Inflammatory Activity of the Extracts of the African Medicinal Plant Ximenia caffra<\/em>. Journal of Analytical Methods in Chemistry<\/em>. 2015, https:\/\/doi.org\/ 10.1155\/2015\/948262
     CrossRef <\/a><\/li>
  10. Kumar N, Singh SK, Lal RK, Sunita Singh Dhawan An insight into dietetic and nutraceutical properties of underutilized legume: Mucuna pruriens<\/em> (L.) DC. Journal of Food Composition and Analysis<\/em>. 2024; 129, 106095. 
    CrossRef <\/a><\/li>
  11. Kumar A, Gupta C, Nair DT, Salunke DM. MP-4 Contributes to Snake Venom Neutralization by Mucuna pruriens Seeds through an Indirect Antibody-mediated Mechanism. Journal of Biological Chemistry<\/em> 2016; 291(21), 11373-84. doi: 10.1074\/jbc.M115.699173
     CrossRef <\/a><\/li>
  12. Boniface F, Washa WB, and Nnungu S. Comparison of nutritional values of Mucuna pruriens<\/em> L. (velvet bean) seeds with the most preferred legume pulses. Food Production, Processing and Nutrition<\/em>.2024; 6, 17. https:\/\/doi.org\/10.1186\/s43014-023-00187-4
    CrossRef <\/a><\/li>
  13. Fadilaturahmah F, Resti R, Putra S. Anti-inflammatory effects of velvet bean (Mucuna pruriens<\/em> L. (DC.), Fabaceae) leaf ethanolic extract against carrageenan in male mice. Journal of Research in Pharmacy<\/em> 2023; 27, 1524-33 http:\/\/dx.doi.org\/10.29228\/jrp.438
     CrossRef <\/a><\/li>
  14. Ganesh MK, Lakshmanan G, Khan MZI, Prakash S. Aging induced testicular damage: analyzing the ameliorative potential of Mucuna pruriens<\/em> seed extract. 3 Biotech<\/em> 2023; 13(6):206. 10.1007\/s13205-023-03618-8
     CrossRef <\/a><\/li>
  15. Kajal K and Pandey RK. Ethnopharmacological uses, phytochemistry and therapeutic potential of Mucuna<\/em> pruriens<\/em>: a comprehensive review on current status of knowledge. Journal of Population Therapeutics and Clinical Pharmacology<\/em> 2024; 31 (3):1184-94. https:\/\/doi.org\/10.53555\/jptcp.v31i3.5101.
     CrossRef <\/a><\/li>
  16. Neshige R, Neshige S. Mucuna beans administered through hydrogen-infused superheated steam in advanced Parkinson’s disease. Clinical parkinsonism & related disorders<\/em> 2024; 10:100252
    CrossRef <\/a><\/li>
  17. Lu K, Lee H, Huang M, Lai S, Ho Y, Chang Y, Chi C. Synergistic Apoptosis-Inducing Antileukemic Effects of Arsenic Trioxide and Mucuna macrocarpa<\/em> Stem Extract in Human Leukemic Cells via a Reactive Oxygen Species-Dependent Mechanism. Evidence-Based Complementary and Alternative<\/em> Medicine<\/em> 2012; 921430. https:\/\/doi.org\/10.1155\/2012\/921430
     CrossRef <\/a><\/li>
  18. Patil RR, Gholave AR, Jadhav JP, Yadav SR, Bapat VA. Mucuna sanjappae<\/em> Aitawade et Yadav: a new species of Mucuna<\/em> with promising yield of anti-Parkinson\u2019s drug L-DOPA. Genetic Resources and Crop Evolution<\/em> 2015; 62, 155\u2013162. https:\/\/doi.org\/10.1007\/s10722-014-0164-8
    CrossRef <\/a><\/li>
  19. Patil RR, Rai SN, Jadhav JP, Singh SP. Mucuna sanjappae<\/em> shows promising anti-Parkinson\u2019s activity by reducing oxidative stress in MPTP induced mouse model. European Journal of Pharmaceutical and Medical Research<\/em> 2016a; 3(11), 452-463.<\/li>
  20. Patil RR, Rane MR, Bapat VA, Jadhav JP.  Phytochemical Analysis and Antioxidant Activity of Mucuna sanjappae<\/em>: A Possible Implementation in the Parkinson\u2019s Disease Treatment. Journal of Pharmaceutical and Medicinal Research<\/em> 2016b; 2(1), 48\u201351.  <\/li>
  21. Aware CB, Patil RR, Vyavahare GD, Gurme ST & Jadhav JP. Ultrasound-Assisted Aqueous Extraction of Phenolic, Flavonoid Compounds and Antioxidant Activity of Mucuna macrocarpa Beans: Response Surface Methodology Optimization. Journal of the American College of Nutrition<\/em> 2019a; 38(4), 364\u2013372. https:\/\/doi.org\/10.1080\/ 07315724. 2018.1524315
     CrossRef <\/a><\/li>
  22. Aware C, Patil R, Bapat V, Gaikwad S, Yadav S, Jadhav J. Evaluation of L-DOPA, proximate composition with in vitro<\/em> anti- the inflammatory and antioxidant activity of Mucuna macrocarpa beans: A future drug for Parkinson\u2019s treatment. Asian Pacific Journal Tropical Biomedicine<\/em> 2017; 7 (12), 1097-1106. https:\/\/doi.org\/10.1016\/j.apjtb.2017.10.012
    CrossRef <\/a><\/li>
  23. Aware C, Patil R, Vyavahare G, Gurav R, Bapat V & Jadhav J. Processing Effect on L-DOPA, In Vitro<\/em> Protein and Starch Digestibility, Proximate Composition, and Biological Activities of Promising Legume: Mucuna macrocarpa. Journal of the American College of Nutrition 2019b; 38(5), 447\u2013456. https:\/\/doi.org\/10.1080\/07315724.2018.1547230
    CrossRef <\/a><\/li>
  24. Rane M, Suryawanshi S, Patil R, et al. Exploring the proximate composition, antioxidant, anti-Parkinson’s and anti-inflammatory potential of two neglected and underutilized Mucuna<\/em> species from India. South African Journal of Botany <\/em>2019; 124:304\u2013310. https:\/\/doi.org\/10.1016\/j.sajb.2019.04.030
     CrossRef <\/a><\/li>
  25. Patil RR, Aware CB, Gaikwad S et al. RP-HPLC Analysis of Anti-Parkinson\u2019s Drug L-DOPA Content in Mucuna<\/em> Species from Indian Subcontinent. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci 2019; 89, 1413\u20131420. https:\/\/doi.org\/10.1007\/s40011-018-01071-9
     CrossRef <\/a><\/li>
  26. Aitawade MM, Yadav SR. Mucuna sanjappae<\/em>, a new species from the north-Western Ghats, India. Kew Bulletin <\/em>2012; 67, 539\u2013543. https:\/\/doi.org\/10.1007\/s12225-012-9369-1
     CrossRef <\/a><\/li>
  27. Sreewardhini S, Sankari D, Vijayalakshmi V, Mangalagowri A, Veeramuthu A. Phytochemical analysis, anti-inflammatory, antioxidant activity of Calotropis gigantea and its therapeutic applications. Journal of Ethnopharmacology<\/em>. 2023; 303, 115963. https:\/\/doi.org\/10.1016\/j.jep.2022.115963
     CrossRef <\/a><\/li>
  28. Yesmin, S., Paul, A., Naz, T. et al. Membrane stabilization as a mechanism of the anti-inflammatory activity of ethanolic root extract of Choi (Piper chaba). Clinical Phytoscience<\/em> 2020; 6, 59. https:\/\/doi.org\/10.1186\/s40816-020-00207-7
     CrossRef <\/a><\/li>
  29. Ola A, Mourad J, Hanen N, Nacim Z, Hichem S, Moncef N. Sulfated polysaccharides from the viscera of Mustelus shark: Characterization and antioxidant, anticoagulant and anti-proliferative activities. Bioactive Carbohydrates and Dietary Fibre<\/em> 2024. 100399. https:\/\/doi.org\/10.1016\/j.bcdf.2023.100399
     CrossRef <\/a><\/li>
  30. Hudz N, Yezerska O, Shanaida M, Hor\u010dinov\u00e1 Sedl\u00e1\u010dkov\u00e1 V, Wieczorek PP. Application of the Folin-Ciocalteu method to the evaluation of Salvia sclarea extracts. Pharmacia <\/em>2019; 66, 209-215. https:\/\/doi.org\/10.3897\/pharmacia.66.e38976
    CrossRef <\/a><\/li>
  31. Chandra S, Khan S, Avula B, Lata H, Yang MH, Elsohly MA, Khan IA. Assessment of total phenolic and flavonoid content, antioxidant properties, and yield of aeroponically and conventionally grown leafy vegetables and fruit crops: a comparative study. Evidence-Based Complementary and Alternative Medicine<\/em>. 2014; 2014, 253875. doi: 10.1155\/2014\/253875.
    CrossRef <\/a><\/li>
  32. Ladhari A, Corrado G, Rouphael Y, Carella F, Nappo GR, Di Marino C, De Marco A, Palatucci D. Chemical, Functional, and Technological Features of Grains, Brans, and Semolina from Purple and Red Durum Wheat Landraces. Foods<\/em>. 2022; 25, 11:1545. doi: 10.3390\/foods11111545
     CrossRef <\/a><\/li>
  33. Soares JC, Zimmermann L, Zendonadi Dos Santos N, Muller O, Pintado M, Vasconcelos MW. Genotypic variation in the response of soybean to elevated CO2. Plant Environment Interactions. 2021; 2, 263-276. doi: 10.1002\/pei3.10065
     CrossRef <\/a><\/li>
  34. Kavitha Chandran CI and Indira G. Quantitative estimation of total phenolic, flavonoids, tannin and chlorophyll content of leaves of Strobilanthes Kunthiana<\/em> (Neelakurinji). Journal of Medicinal Plants Studies<\/em> 2016; 4 (4), 282-286. https:\/\/www.plantsjournal.com\/archives\/ 2016\/vol4issue4\/PartD\/4-4-4-759.pdf<\/li>
  35. Alam F, Us Saqib QN. Pharmacognostic study and development of quality control parameters for fruit, bark and leaf of Zanthoxylum armatum (Rutaceae). Ancient Science of Life<\/em>, 2015; 34(3), 147-55. doi: 10.4103\/0257-7941.157159.
     CrossRef <\/a><\/li>
  36. Najmi A, Javed SA, Al Bratty M, Alhazmi HA. Modern Approaches in the Discovery and Development of Plant-Based Natural Products and Their Analogues as Potential Therapeutic Agents. Molecules<\/em>. 2022; 27(2), 349. https:\/\/doi.org\/10.3390\/molecules27020349
     CrossRef <\/a><\/li>
  37. Nedeljkovi\u0107 N, Dobri\u010di\u0107 V, Bo\u0161kovi\u0107 J, Vesovi\u0107 M, Bradi\u0107 J, An\u0111i\u0107 M, Ko\u010dovi\u0107 A, Jeremi\u0107 N, Novakovi\u0107 J, Jakovljevi\u0107 V, Vuji\u0107 Z, Nikoli\u0107 M. Synthesis, and Investigation of Anti-Inflammatory Activity of New Thiourea Derivatives of Naproxen. Pharmaceuticals (Basel). 2023; 16(5):666. doi: 10.3390\/ph16050666
     CrossRef <\/a><\/li>
  38. Bindu S, Mazumder S, Bandyopadhyay U. Non-steroidal anti-inflammatory drugs (NSAIDs) and organ damage: A current perspective. Biochemical Pharmacology<\/em>, 2020; 180, 114147. doi: 10.1016\/j.bcp.2020.114147.
     CrossRef <\/a><\/li>
  39. Pradhan B, Jang-Seu K Biological activity of algal derived carrageenan: A comprehensive review in light of human health and disease. International Journal of Biological Macromolecules. <\/em>2023, 238, 124085. https:\/\/doi.org\/10.1016\/j.ijbiomac.2023.124085
    CrossRef <\/a><\/li>
  40. Patil KR, Mahajan UB, Unger BS, Goyal SN, Belemkar S, Surana SJ, Ojha S, Patil CR. Animal Models of Inflammation for Screening of Anti-inflammatory Drugs: Implications for the Discovery and Development of Phytopharmaceuticals. International Journal of Molecular Science<\/em>. 2019; 20, 4367. doi: 10.3390\/ijms20184367.
     CrossRef <\/a><\/li>
  41. Mondello S, Hayes R Biomarkers Handbook of Clinical Neurology<\/em> 2015; 127, Pages 245-265. https:\/\/doi.org\/10.1016\/B978-0-444-52892-6.00016-7
     CrossRef <\/a><\/li>
  42. Srinivas US, Tan BWQ, Vellayappan BA, Jeyasekharan AD. ROS and the DNA damage response in cancer. Redox Biology<\/em> 2019; 25, 101084, 10.1016\/j.redox.2018.101084
     CrossRef <\/a><\/li>
  43. Oluwole O, Fernando WMAD, Lumanlan J, Ademuyiwa O, Jayasena V. Role of phenolic acid, tannins, stilbenes, lignans and flavonoids in human health \u2013 a review. International Journal of Food Science & Technology<\/em>; 2022, 57, 6326-6335. https:\/\/doi.org\/10.1111\/ijfs.15936.
     CrossRef <\/a><\/li>
  44. Verma P, Sen R, Bamanna A, Elhindawy M, Nagpal K, Krishnan V. Structural chemistry to therapeutic functionality: A comprehensive review on proanthocyanidins. Biocatalysis and Agricultural Biotechnology<\/em> 2024; 55, 102963.
     CrossRef <\/a><\/li>
  45. Salinas-S\u00e1nchez DO, Jim\u00e9nez-Ferrer E, S\u00e1nchez-S\u00e1nchez V, Zamilpa A, Gonz\u00e1lez-Cortazar M, Tortoriello J, Herrera-Ruiz M. Anti-Inflammatory Activity of a Polymeric Proanthocyanidin from Serjania schiedeana<\/em>. Molecules<\/em> 2017; 22(6), 863. https:\/\/doi.org\/10.3390\/molecules22060863
     CrossRef <\/a><\/li>
  46. Limtrakul P, Yodkeeree S, Pitchakarn P, Punfa W. Anti-inflammatory effects of proanthocyanidin-rich red rice extract via suppression of MAPK, AP-1 and NF-\u03baB pathways in Raw 264.7 macrophages. Nutrition Research and Practice<\/em> 2016; 10(3), 251-258. https:\/\/doi.org\/10.4162\/ nrp.2016.10.3.251
     CrossRef <\/a><\/li>
  47. Abdulwaliyu I, Arekemase SO, Adudu JAA, Batari ML, Egbule MN, Okoduwa.SIR. Investigation of the medicinal significance of phytic acid as an indispensable anti-nutrient in diseases, Clinical Nutrition Experimental <\/em>2019; 28, 42-61. https:\/\/doi.org\/10.1016\/ j.yclnex.2019.10.002
     CrossRef <\/a><\/li>
  48. Lv Yuqiang, Zhang Zheng, Hou Lin, Zhang Li.  Phytic acid attenuates inflammatory responses and the levels of NF-\u03baB and p-ERK in MPTP-induced Parkinson\u2019s disease model of mice. Neuroscience Letters <\/em>2015; <\/em>597, 132-136. https:\/\/doi.org\/10.1016\/j.neulet.2015.04.040
     CrossRef <\/a><\/li>
  49. Chen Y, Yuan W, Xu Q, Reddy M. Neuroprotection of phytic acid in Parkinson\u2019s and Alzheimer\u2019s disease, Journal of Functional Foods 2023; <\/em>110, 105856. https:\/\/doi.org\/10.1016\/j.jff.2023.105856
     CrossRef <\/a><\/li>
  50. Wijesekara T, Luo J, Xu B. Critical review on anti-inflammation effects of saponins and their molecular mechanisms. Phytotherapy Research<\/em>. 2024; 38(4), 2007-2022. doi: 10.1002\/ptr.8164.
     CrossRef <\/a><\/li>
  51. Park M, Cho H, Jung H, Lee H, Hwang K. Antioxidant and Anti-Inflammatory Activities of Tannin Fraction of the Extract from Black Raspberry Seeds Compared to Grape Seeds. Journal of food Biochemistry <\/em>2014; 38 (3), 259\u2013270. https:\/\/doi.org\/10.1111\/jfbc.12044
     CrossRef <\/a><\/li><\/ol>\n\n\n\n

    Abbreviations<\/strong><\/p>\n\n\n\n

    BSA: Bovine serum albumin<\/p>\n\n\n\n

    ELISA:\nEnzyme-linked immunosorbent assay<\/p>\n\n\n\n

    HRBC:Human Red Blood Cell <\/p>\n\n\n\n

    IL-10: Interleukin 10<\/p>\n\n\n\n

    L-DOPA: L-3,4-dihydroxyphenylalanine<\/p>\n\n\n\n

    MPTP:\nN-methyl-4-phenyl-l,2,3,6-tetrahydropyridine<\/p>\n\n\n\n

    Mucuna sanjappae <\/em>water\nextract<\/p>\n\n\n\n

    NSAIDs: Non-steroidal\nanti-inflammatory drugs<\/p>\n\n\n\n

    NF-\u03baB: Nuclear\nfactor-\u03baB <\/p>\n\n\n\n

    ORAC assay: Oxygen radical absorbance\ncapability<\/p>\n\n\n\n

    p-ERK:\nPhosphorylated extracellular signal-related kinase<\/p>\n\n\n\n

    PD: Parkinson\u2019s disease<\/p>\n\n\n\n

    RBC: Red blood cell<\/p>\n\n\n\n

    TNF-\u03b1: Tumor Necrosis Factor alpha<\/p>\n","protected":false},"excerpt":{"rendered":"

    Introduction Inflammation can be defined as essential response exhibited by […]<\/p>\n","protected":false},"author":15,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[115],"tags":[],"class_list":["post-59226","post","type-post","status-publish","format-standard","hentry","category-vol17no2"],"_links":{"self":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/59226","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/users\/15"}],"replies":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/comments?post=59226"}],"version-history":[{"count":5,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/59226\/revisions"}],"predecessor-version":[{"id":60848,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/59226\/revisions\/60848"}],"wp:attachment":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/media?parent=59226"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/categories?post=59226"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/tags?post=59226"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}