{"id":51964,"date":"2023-09-30T10:42:20","date_gmt":"2023-09-30T10:42:20","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=51964"},"modified":"2023-10-07T11:24:53","modified_gmt":"2023-10-07T11:24:53","slug":"isolation-and-evaluation-of-cytotoxic-anti-inflammatory-anti-ulcer-activity-of-methanolic-extract-of-ceriops-decandra-leaves","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol16no3\/isolation-and-evaluation-of-cytotoxic-anti-inflammatory-anti-ulcer-activity-of-methanolic-extract-of-ceriops-decandra-leaves\/","title":{"rendered":"Isolation and Evaluation of Cytotoxic, Anti-Inflammatory, Anti-Ulcer Activity of Methanolic Extract of Ceriops decandra leaves"},"content":{"rendered":"\n

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

Only 124 nations and regions have at least one natural mangrove species, with Asia having the most mangrove acreage. Mangrove is essential to the ecosystem by protecting the seashore and providing a habitat for marine and terrestrial organisms. Mangroves grow in the intertidal zone of extreme environmental conditions like inundation, anaerobic soil, strong wind, and high salinity1<\/sup>. Because of their extreme morphological and physiological adaptions, mangroves have a higher chance of producing enhanced bioactive compounds. Traditionally, it is used to treat gastrointestinal disorders, inflammation, cancer and infection2<\/sup>. <\/p>\n\n\n\n

Ceriops decandra<\/em> is a mangrove plant of the family Rhizophoraceae. It is found throughout tropical Asian countries like Bangladesh and Eastern Africa, Queensland and Madagascar. It is rich in phytochemicals such as protein, cumarin, phenols (catechin, procyanidins), triterpenoids (lupeol, \u03b1-amyrin, oleanolic acid, ursolic acid), flavonoids, saponins, glycosides, alkaloids diterpenoids (ceriopsin A-G), and tannins3<\/sup>. <\/p>\n\n\n\n

Despite severe cardiovascular and gastrointestinal side effects, NSAIDs are widely used as analgesic and anti-inflammatory agents. COX 2 selective analgesics such as Celecoxib are safe for Gastro intestinal tract than a nonselective agent. The search for newer and safer analgesic and anti-inflammatory agents continues for this4<\/sup>. The lack of knowledge regarding Ceriops decandra<\/em> methanolic extracts cytotoxic, anti-inflammatory, and anti-ulcer properties in relation to Bangladeshi species inspired this work. These findings were accompanied by histological analysis of the anti-inflammatory and anti-ulcer actions as well as chemical component separation.<\/p>\n\n\n\n

Materials and method<\/strong><\/p>\n\n\n\n

Plant materials<\/strong><\/p>\n\n\n\n

The fresh leaves of Ceriops Decandra<\/em> were collected from the Sundarbans mangrove forest in June-July. The plant’s identity was confirmed by the Bangladesh National Herbarium, where a voucher specimen was kept (Voucher no- DACB-85929).<\/p>\n\n\n\n

Chemicals and drugs<\/strong><\/p>\n\n\n\n

Eskayef Bangladesh Limited provided the omeprazole and ibuprofen that were collected. Carrageenan was acquired from Sigma Aldrich compounds in Germany, and analytical-grade Petroleum ether, ethyl acetate, and other compounds were purchased from Merck in Germany.<\/p>\n\n\n\n

Extraction of plant materials<\/strong><\/p>\n\n\n\n

2.5 kg of fresh C. decandra <\/em>leaves were separated, shade-dried, and ground into a coarse powder. The coarse powder was then extracted with methanol using a Soxhlet apparatus at Jahangirnagar University and vacuum-dried.<\/p>\n\n\n\n

Extraction and isolation from Ceriops decandra<\/em><\/strong><\/p>\n\n\n\n

For approximately six\nmonths, 40 g of crude methanolic extract were stored in a refrigerator at 3-5\n\u00b0C. The substance underwent a natural separation process, resulting in the\nformation of two distinct layers: a lower layer characterized by its sticky and\nresinous properties, and an upper layer consisting of a solid component. In\nthis study, the solid component was utilized to extract and separate Lupeol.\nThe solid component was separated into a fraction that is soluble in petroleum\nether using a separating funnel. The organic fraction, which is soluble in\npetroleum ether, was subsequently dried at room temperature, resulting in a\nyield of 9 grams. <\/p>\n\n\n\n

The petroleum ether\nfraction was subjected to fractionation using silica column chromatography (CC)\nutilizing a gradient elution system consisting of petroleum ether, ethyl\nacetate, and methanol in varying proportions (ranging from 99:1 to 20:80, v\/v).\nTherefore, the entirety of the dissolved fraction of petroleum ether (9 g) was\npartitioned into 75 fractions, each containing a solvent system of 20 ml. <\/p>\n\n\n\n

The impure, colorless crystalline substance was formed through the process of evaporating the solvents (85:15-60:40, v\/v) of fractions 22 to 31, which consisted of Petroleum ether and ethyl acetate. The production of compound 1 (30 mg, Rf=0.55) involved the repeated washing of crystal bodies with n-hexane (100:0, v\/v) and subsequent recrystallization using a mixture of n-hexane and ethyl acetate (50:50, v\/v). The resulting crystals were pure.<\/p>\n\n\n\n

Modified Kupchan Partitioning Method<\/strong><\/p>\n\n\n\n

Crude methanolic extract (5gm) was fractionated using petroleum ether (100ml, 3 times). Again, the aqueous fraction from this extraction was fractionated using ethyl acetate. The ethyl acetate soluble fraction (4.8 gm) and aqueous fraction (1.75 gm) were then used for the brine shrimp cytotoxicity assay. <\/p>\n\n\n\n

Brine shrimp lethality bioassay<\/strong><\/p>\n\n\n\n

The lethality test on brine shrimp was used to determine the plant extract’s cytotoxic activity. The concentrations of the dried extract preparations were serially diluted to range from 5 g\/ml to 320 g\/ml. Positive control was provided by the conventional medication vincristine sulphate (VS), whose concentrations ranged from 0.312 g\/ml to 5 g\/ml. The brine shrimp\u2019s (Artemia salina<\/em>) eggs were obtained from a neighborhood aquarium store in Dhaka, Bangladesh, and they were incubated in brine for 24 hours with a continual supply of oxygen. After 24 hours, the number of alive nauplii out of 10 were counted. These experiments were conducted on triplets. The lethal concentration (LC50) required to eliminate 50% of the nauplii was calculated using the application “Microsoft Excels 2007”9<\/sup>.<\/sub><\/p>\n\n\n\n

Anti-inflammatory Activity<\/strong><\/p>\n\n\n\n

Experimental Animal <\/strong><\/p>\n\n\n\n

This experiment was designed following the previously described method discussed by Begum et al.<\/em> 5,6<\/sup>. Male Albino wistar rats weighing 120 and 150 g were collected from Jahangirnagar University’s pharmacy department. They were kept in groups of six rats in polypropylene cages, with a room temperature of 25 2 \u00b0C and a 12-hour light\/dark cycle. The rats were given a week of acclimation before the studies, during which they had unrestricted access to free standard laboratory animal food and water. They went without eating the night before the test. All procedures used the anesthetic isoflurane (5% in 100% oxygen) to reduce preventable harm. The principles and guidelines outlined in the Guide for the Care and Use of Laboratory Animals served as the cornerstone for the treatment of animals and research procedures (NIH document no. 85-23; updated in 1985).<\/p>\n\n\n\n

Induction of Inflammation in experimental animals<\/strong><\/p>\n\n\n\n

The\nreference standard utilized was ibuprofen (20 mg\/kg), and the anti-inflammatory\naction was carried out as advised by Winter et al. by injecting carrageenan as\nan edematogenic agent on adult albino rats.<\/p>\n\n\n\n

The following five groups, each with six rats, were formed:<\/p>\n\n\n\n\n\n\n\n\n\n\n
\n

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

\n

Description<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Group I<\/p>\n<\/td>\n

\n

oral administration of 0.5ml distilled water<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

oral administration of 0.5ml distilled water and 0.1ml of Carageenan injection (Carageenan control, CC)<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

Oral administration of Ibuprofen (20 mg\/kg) and 0.1ml of Carageenan injection (IP 20)<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

250 mg\/kg methanolic extract of C.decendra <\/em>and 0.1ml of Carageenan injection  (MECD-250)<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

500 mg\/kg methanolic extract of C.decendra <\/em>and 0.1ml of Carageenan injection (MECD-500)<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

The extracts were given orally to the treatment group as a suspension containing 2 to 3 drops of tween 80 at 250 and 500 mg\/kg doses, respectively. Thirty minutes before injecting carrageenan (0.1 ml of 1% w\/v solution of distilled water) (0.5 ml), all test samples were administered orally. Carrageenan was injected into the sub-planter area of the left paw of each rat. However, the normal control group received no carrageenan. Slide callipers were used to gauge and record the paw swelling every hour. The percentage inhibition of paw edema was calculated after three hours.<\/p>\n\n\n\n

The formula for the percentage of\ninhibition in control and the increase in paw thickness is as follows:<\/p>\n\n\n\n

Increase of paw thickness in control or treatment group = PC or PT= Pt \u2013 Po<\/p>\n\n\n\n

Percentage of inhibition in paw thickness in the treatment group<\/p>\n\n\n\n

\"\"<\/figure>\n\n\n\n

Where, Pt = paw thickness at time\nt, <\/p>\n\n\n\n

Po = initial paw thickness, <\/p>\n\n\n\n

PC = Increase in thickness of paw\nof the control group and <\/p>\n\n\n\n

PT = Increase in thickness of paw of the treatment group.<\/p>\n\n\n\n

Anti-ulcer activity<\/strong><\/p>\n\n\n\n

Induction of Ulcer<\/strong><\/p>\n\n\n\n

This experiment was performed following Begum et al. 6,7<\/sup>. Five groups of animals were not allowed to access food for a day, other than water, for two hours before the test. The rats were kept apart in their cages to avoid coprophagy throughout the fasting phase. Gastric ulcers were induced in the test sample (MECD at doses of 250 and 500 mg\/kg p.o.) and standard (omeprazole at doses of 20 mg\/kg, p.o.) using ethanol-acid after 30 minutes of pretreatment. Under isoflurane, 5% in 100% oxygen anesthesia, these rats were euthanized 90 minutes later, and their stomachs were immediately removed. The stomachs were sliced open along the larger curvature, removed, cleaned with distilled water, and stored in 10% buffered formalin. The gastrointestinal mucosa was examined under a microscope for ulcers, and the ulcers were graded as follows:<\/p>\n\n\n\n\n\n\n\n\n\n\n\n
\n

Tissue architecture<\/strong><\/p>\n<\/td>\n

\n

Ulcer score<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

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

\n

Red coloration<\/p>\n<\/td>\n

\n

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

\n

Spot ulcer<\/p>\n<\/td>\n

\n

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

\n

Hemorrhagic streak<\/p>\n<\/td>\n

\n

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

\n

Ulcers<\/p>\n<\/td>\n

\n

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

\n

Perforation<\/p>\n<\/td>\n

\n

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

The average ulcer score for each animal is represented by the ulcer index. The percentage of ulcer prevention was calculated using the formula below: <\/p>\n\n\n\n

\"\"<\/figure>\n\n\n\n

The animals were grouped into five groups, each with six rats:<\/p>\n\n\n\n\n\n\n\n\n\n\n
\n

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

\n

Description<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Group I<\/p>\n<\/td>\n

\n

Oral administration of 5 ml\/kg\/day distilled water (Normal Control)<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

Oral administration of 5 ml\/kg\/day distilled water and 25 ml per kg of 0.3M HCl in 60 % ethanol (Negative control)<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

Oral administration of 20 mg\/kg\/day omeprazole and 25 ml per kg of 0.3M HCl in 60 % ethanol<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

Oral administration of 250 mg\/kg methanolic extract of C.decendra <\/em>25 ml per kg of  0.3M HCL in 60 % ethanol<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

Oral administration of 500 mg\/kg methanolic extract of C.decendra <\/em>25 ml per kg of 0.3M HCL in 60 % ethanol<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Histological investigation<\/strong><\/p>\n\n\n\n

Carrageenan-induced paw tissues\nwere collected for this study 3 hours after the induction and kept in freshly\nmade 10% neutral buffered formalin for at least 24 hours. The specimens for the\nanti-ulcer study were kept in a 10% natural buffer. <\/strong>Following that, tissue slices were\nparaffin-embedded. Using a microtome, the samples were cut into cross-sections\nstained with hematoxylin and eosin (H and E) and mounted on Canada balsam. Each\nsegment was inspected under a light microscope, and any histopathological\nalterations, such as edema, erosion, infiltration, and inflammation, were\nphotographed using an Olympus photomicroscope.<\/p>\n\n\n\n

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

The values are tabulated as mean \u00b1 S.E.M, and statistical significance between treated and control groups was analyzed using One-way ANOVA, where P<0.05 was considered statistically significant.<\/p>\n\n\n\n

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

Characterization of isolated compounds<\/strong><\/p>\n\n\n\n

Compound 1 is a pentacyclic triterpene. It was a white powder. Seven tertiary methyl singlets and one secondary hydroxyl group were seen in the 1<\/sup>H NMR spectra. Additionally, it displayed olefinic protons at 4.71 and 4.59. The compound’s 1<\/sup>H NMR spectra showed signals at \u03b4 4.71 and \u03b4 4.59, both of which were focused on the furan exomethylene group, as one proton doublets with 2.4 Hz coupling constants and one proton double quartets with 1.2 Hz and 2.4 Hz coupling constants, respectively. A secondary carbinol group may be given to the signals at \u03b4 3.21 that appeared as a doublet of doublets with coupling constants of 11.2 Hz and 5.2 Hz. A wide doublet at 1.66 with a coupling constant of 1.2 Hz was also visible in the spectrum and was attributed to a methine group. Six tertiary methyl singlets could be seen in the resonance signals at 0.79, 0.81, 0.85, 0.97, 0.99, and 1.05. These results closely matched those that had previously been published for a common pentacyclic triterpenoid called lupeol7<\/sup>.<\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Scheme 1: Lupeol<\/strong><\/p>\n

<\/p>\n

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

13<\/sup>C NMR value: (\u03b4 178.2, C-28), (\u03b4 151.6, C-20), (\u03b4 126.0, C-29), (\u03b4 178.2, C-28), (\u03b4 79.1, C-3), (\u03b4 55.8, C-17), (\u03b4 56.1, C-5), (\u03b4 51.1, C-9), (\u03b4 50.0, C-19), (\u03b4 46.8, C-18), (\u03b4 41.8, C-14), (\u03b4 41.0, C-24), (\u03b4 40.0, C-4), (\u03b4 40.1, C-1), (\u03b4 37.9, C-13), (\u03b4 38.0, C-10), (\u03b4 37.5, C-22), (\u03b4 178.2, C-28), (\u03b4 34.8, C-7), (\u03b4 32.5, C-16), (\u03b4 31.0, C-15), (\u03b4 29.9, C-21), (\u03b4 28.1, C-23), (\u03b4 28.4, C-2), (\u03b4 26.0, C-12), (\u03b4 21.0, C-11), (\u03b4 19.0, C-30), (\u03b4 18.7, C-6), (\u03b4 16.0, C-25), (\u03b4 15.2, C-27)8<\/sup>.<\/p>\n\n\n\n

Brine Shrimp Cytotoxicity<\/strong><\/p>\n\n\n\n

The effect of methanolic extract of Ceriops\ndecandra <\/em>as well as Vincristine sulphate on brine shrimp nauplii has been\ntabulated and graphically represented. <\/p>\n\n\n\n

Table 1: Effect of aqueous and petroleum ether fraction of Ceriops decandra <\/em>as well as Vincristine sulphate on brine shrimp nauplii<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Ceriops decandra<\/em><\/strong> leaf extract<\/strong><\/p>\n<\/td>\n

\n

Vincristine Sulphate<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Concentration, C (\u00b5g\/ml)<\/p>\n<\/td>\n

\n

Log C<\/p>\n<\/td>\n

\n

% mortality<\/p>\n

(aqueous fraction)<\/p>\n<\/td>\n

\n

% mortality<\/p>\n

(Petroleum ether fraction)<\/p>\n<\/td>\n

\n

Concentration, CV<\/sub> (\u00b5g\/ml)<\/p>\n<\/td>\n

\n

Log CV<\/sub><\/p>\n<\/td>\n

\n

% mortality<\/p>\n<\/td>\n<\/tr>\n<\/thead>\n

\n

5<\/p>\n<\/td>\n

\n

0.693<\/p>\n<\/td>\n

\n

25<\/p>\n<\/td>\n

\n

5<\/p>\n<\/td>\n

\n

0.312<\/p>\n<\/td>\n

\n

-0.505<\/p>\n<\/td>\n

\n

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

\n

10<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

30<\/p>\n<\/td>\n

\n

15<\/p>\n<\/td>\n

\n

0.625<\/p>\n<\/td>\n

\n

-0.204<\/p>\n<\/td>\n

\n

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

\n

20<\/p>\n<\/td>\n

\n

1.301<\/p>\n<\/td>\n

\n

35<\/p>\n<\/td>\n

\n

20<\/p>\n<\/td>\n

\n

1.25<\/p>\n<\/td>\n

\n

0.096<\/p>\n<\/td>\n

\n

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

\n

40<\/p>\n<\/td>\n

\n

1.602<\/p>\n<\/td>\n

\n

40<\/p>\n<\/td>\n

\n

30<\/p>\n<\/td>\n

\n

2.50<\/p>\n<\/td>\n

\n

0.397<\/p>\n<\/td>\n

\n

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

\n

80<\/p>\n<\/td>\n

\n

1.903<\/p>\n<\/td>\n

\n

50<\/p>\n<\/td>\n

\n

45<\/p>\n<\/td>\n

\n

5.0<\/p>\n<\/td>\n

\n

0.698<\/p>\n<\/td>\n

\n

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

\n

160<\/p>\n<\/td>\n

\n

2.204<\/p>\n<\/td>\n

\n

55<\/p>\n<\/td>\n

\n

55<\/p>\n<\/td>\n

\n

 <\/p>\n<\/td>\n

\n

 <\/p>\n<\/td>\n

\n

 <\/p>\n<\/td>\n<\/tr>\n

\n

320<\/p>\n<\/td>\n

\n

2.505<\/p>\n<\/td>\n

\n

65<\/p>\n<\/td>\n

\n

70<\/p>\n<\/td>\n

\n

 <\/p>\n<\/td>\n

\n

 <\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n

Table 2: LD50 Value of extracts Ceriops decandra <\/em>aqueous and petroleum ether part as well as vincristine sulphate<\/strong><\/p>\n\n\n\n\n\n\n\n\n
\n

Sample<\/strong><\/p>\n<\/td>\n

\n

LC50<\/sub> (\u03bcg\/ml)<\/strong><\/p>\n<\/td>\n

\n

Regression line equation<\/strong><\/p>\n<\/td>\n

\n

R2<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Ceripos decendra<\/em> (Aqueous fraction)<\/p>\n<\/td>\n

\n

1.93<\/p>\n<\/td>\n

\n

y = 21.899x + 7.7936<\/p>\n<\/td>\n

\n

0.9831<\/p>\n<\/td>\n<\/tr>\n

\n

Ceripos decendra<\/em><\/p>\n

(petroleum ether fraction)<\/p>\n<\/td>\n

\n

2.04<\/p>\n<\/td>\n

\n

y = 35.513x – 22.575<\/p>\n<\/td>\n

\n

0.9822<\/p>\n<\/td>\n<\/tr>\n

\n

Vincristine sulphate<\/p>\n<\/td>\n

\n

0.02<\/p>\n<\/td>\n

\n

y = 76.485x + 48.627<\/p>\n<\/td>\n

\n

0.9934<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\n\n

Figure 1<\/strong>: Plot of log C % mortality of Ceriops decandra <\/em>petroleum ether and aqueous fraction.<\/em><\/strong><\/p>\n

<\/p>\n

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

Anti-inflammatory activity of Ceriops decandra<\/em><\/strong><\/p>\n\n\n\n

In this\nstudy, animals given methanolic extract (250 and 500 mg\/kg, p.o.) and ibuprofen\n(20 mg\/kg, p.o.) for 1 to 3 hours showed considerably reduced paw thickness and\nredness than the carrageenan control group. \nAfter the investigation, it was found that when compared to the\ncarrageenan control group at 3 hours after carrageenan injection, oral\nadministration of methanolic extract of 250mg\/kg and 500mg\/kg reduced paw edema\nby 62.5% and 87.5%, respectively.<\/p>\n\n\n\n

Table 3: Anti-inflammatory effect of Ceriops decandra<\/em> methanolic extract on the carrageenan induced changes in paw thickness in experimental rats<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
\n

Groups<\/strong><\/p>\n

Dose mg\/kg<\/strong><\/p>\n<\/td>\n

\n

NC<\/strong><\/p>\n

 <\/p>\n<\/td>\n

\n

CC<\/strong><\/p>\n

 <\/p>\n<\/td>\n

\n

IP-20<\/strong><\/p>\n

(20mg\/kg Ibuprofen)<\/strong><\/p>\n<\/td>\n

\n

MECD-250<\/strong><\/p>\n

(250mg\/kg extract)<\/strong><\/p>\n<\/td>\n

\n

MECD-500<\/strong><\/p>\n

(500mg\/kg extract)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Initial paw thickness (cm)<\/p>\n<\/td>\n

\n

0.48\u00b10.004<\/p>\n<\/td>\n

\n

0.49\u00b10.005c<\/sup><\/p>\n<\/td>\n

\n

0.46\u00b10.003b**<\/sup><\/p>\n<\/td>\n

\n

0.49\u00b10.003b<\/sup><\/p>\n<\/td>\n

\n

0.48\u00b10.004c<\/sup><\/p>\n<\/td>\n<\/tr>\n

\n

Paw thickness (cm) after 1h<\/p>\n<\/td>\n

\n

0.48\u00b10.004<\/p>\n<\/td>\n

\n

0.67\u00b10.004a**<\/sup><\/p>\n<\/td>\n

\n

0.57\u00b10.006b**<\/sup><\/p>\n<\/td>\n

\n

0.64\u00b10.004b<\/sup><\/p>\n<\/td>\n

\n

0.53\u00b10.007b**<\/sup><\/p>\n<\/td>\n<\/tr>\n

\n

Paw thickness (cm) after 2h<\/p>\n<\/td>\n

\n

0.48\u00b10.002<\/p>\n<\/td>\n

\n

0.70\u00b10.007a**<\/sup><\/p>\n<\/td>\n

\n

0.54\u00b10.003b**<\/sup><\/p>\n<\/td>\n

\n

0.61\u00b10.007b*<\/sup><\/p>\n<\/td>\n

\n

0.52\u00b10.004 b**<\/sup><\/p>\n<\/td>\n<\/tr>\n

\n

Paw thickness (cm) after 3h<\/p>\n<\/td>\n

\n

0.48\u00b10.003<\/p>\n<\/td>\n

\n

0.73\u00b10.004a**<\/sup><\/p>\n<\/td>\n

\n

0.48\u00b10.003b**<\/sup><\/p>\n<\/td>\n

\n

0.58\u00b10.003b**<\/sup><\/p>\n<\/td>\n

\n

0.51\u00b10.004 b**<\/sup><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Each\nvalue is presented as Mean SEM (n=6). NC stands for “Healthy\nControl,” CC stands for “Carrageenan Control,” IP-20 stands for\n“Ibuprofen 20 mg\/kg\/day,” MECD-250 stands for “Methanolic\nExtract of C. Decendra 250 mg\/kg,” and MECD-500 stands for “Methanolic\nExtract of C. Decendra<\/em> 500 mg\/kg.” Dunnett’s multiple comparison\ntest (*p 0.05 and **p0.01) is used after one way analysis of variance to show\nstatistically significant differences from the respective group. uses one-way\nanalysis of variance to demonstrate that there is no statistically significant\ndifference from the corresponding group, followed by Dunnett’s multiple\ncomparison test (p > 0.05). when contrasted with the control (carrageenan)\nand the control (normal). <\/p>\n\n\n\n

Effect\nof methanolic extract of Ceriops decendra<\/em>\non the carrageenan induced paw edema<\/strong><\/p>\n\n\n\n

In the healthy control (2a)\nthere was no inflammation or erythema observe. However, in Carrageenan control (2b)\nsignificant erythema and edema were observed. In case of Ibuprofen 20 mg\/kg (2c),\nminor erythema and edema is seen. The paw of C. decendra 250 mg\/kg (2d) methanolic\nextract treated rat shows mild erythema and edema and 500 mg\/kg methanolic\nextract of C. decendra (2e) treated rat shows comparatively less edema and\nerythema.<\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure <\/strong>2<\/strong>: Effect of methanolic extract of Ceriops decandra<\/em> on carrageenan induced paw edema after 3 h.<\/strong><\/p>\n

<\/p>\n

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

Histopathological Evaluations of\nPaw Tissue<\/strong><\/p>\n\n\n\n

When paw tissue was\nexamined histopathologically, it can be deduced that tissues from normal\ncontrols (3a) have few leukocytes. In contrast, tissues from\ncarrageenan-induced rat paws (3b) have a large number of inflammatory cells. On\nthe other hand, tissues from rat paws treated with the standard drug Ibuprofen\n(3c) and C. decandra<\/em> methanolic extract at dosages of 250 (3d) and 500\nmg\/kg (3e) exhibit only a moderate to low level of inflammatory cell\ninfiltration.<\/p>\n\n\n\n\n\n
\"\"<\/td>\nFigure 3<\/strong>: Histopathological Evaluations of Paw Tissue of a) Normal control b) Carrageenan control c) 20mg\/kg Ibuprofen d) Methanolic extract of Ceriops decandra <\/em>leaves at dose 250mg\/kg <\/strong>

<\/p>\n

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

Ethanol-Induced Gastric Ulcer<\/strong><\/p>\n\n\n\n

After dissecting the stomachs in the greater curvature, the ethanol-treated rat tissues (4b) have thick, dark red and black lesions in the glandular portion. As compared to normal control (4a), Omeprazole (20mg\/kg) (4c), Methanolic extract at 250mg\/kg (4d) and 500 mg\/kg (4e) treated rat tissues have much better ulcer protection with a significant protection index of 63.6 %, 45.5% and 59.1% respectively.<\/p>\n\n\n\n\n\n
\"\"<\/td>\nFigure <\/strong>4<\/strong>: Gross appearance of the gastric mucosa: a) Normal control (healthy stomach) b) Ethanol control (developed significant ulceration with mucosal injury) c) Omeprazole 20 mg\/kg (less area of gastric ulceration) <\/strong>

<\/p>\n

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

Histological\nEvaluation of Gastric Lesions<\/strong><\/p>\n\n\n\n

Figure 5a <\/strong>shows a section of normal control which only received distilled water. On the contrary, the ethanol control group 5b shows ulceration with broken mucosal barrier and migration of numerous inflammatory cells. However, omeprazole (20 mg\/kg) (5c) administration changed histopathology significantly, with minimal hemorrhage presence and intact mucosal lining. Methanolic extract of C. decendra <\/em>at 250 mg\/kg (5d) showed moderate regeneration of gastric mucosa, while 500 mg\/kg (5e) showed significant regeneration. <\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 5<\/strong>: Histological Evaluation of Gastric Lesions a) Normal control, b) Ethanol control, c) Omeprazole 20 mg\/kg, d) Methanolic extract of C. decendra (<\/em>250 mg\/kg), e) Extract of C. decendra (<\/em>500 mg\/kg).<\/strong><\/p>\n

<\/p>\n

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

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

Based on the 1<\/sup>H and 13<\/sup>C NMR values for each proton and carbon that were determined and provided in the results and discussions lupeol skeleton for compound 1, the lack of a hydroxyl group, and the appearance of a carbonyl group in the 13<\/sup>C NMR spectrum data of 1. As a result, structure 1 was determined to be that of the well-known chemical lupeol, which was supported by the physical and spectral information in the literature9<\/sup>.<\/p>\n\n\n\n

In this\nwork, a methanolic extract of Ceriops decandra<\/em> showed\na dose-dependent anti-inflammatory activity in paw edema caused by carrageenan.\nThe outcome is nearly as significant as the standard drug Ibuprofen. The\nsuppression of mediators such as prostaglandins, leukotrienes, histamine,\nserotonin, and bradykinins, which cause pain and fever, may cause\nanti-inflammatory action10<\/sup>. <\/p>\n\n\n\n

The\nhistological analysis of the hind paw tissue in our study showed that the\nmethanolic extract of Ceriops decandra<\/em>\nreduced the massive influx and buildup of inflammatory cells in the paw tissue\nafter carrageenan induction in a dose-dependent manner. TNF-a is a crucial\nelement of inflammation. Among other things, it can trigger the release of\npro-inflammatory cytokines and the activation and chemotaxis of leukocytes.\nLeukocyte influx-induced inflammation can be avoided by reducing the release of\nmediators such MDA, CRP, NO, MPO, and pro-inflammatory cytokines (TNF-a and\nIL-1b)11<\/sup>.<\/p>\n\n\n\n

Since\nethanol may permeate the stomach mucosa, the mucosal barrier will break down,\nincreasing the permeability of sodium and water and causing calcium ions to\naccumulate. Additionally, it increases the production of superoxide ions and\nhydroperoxyl free radicals, which results in stomach mucosal ulcers10<\/sup>.\nThe stomach cryoprotection through the anti-oxidation mechanism and the\ndecreased release of gastric acid and prostaglandins may all contribute to the\nanti-ulcer effects of the methanolic extract of Ceriops decandra<\/em>11<\/sup>. The anti-ulcer effect was\ndose-dependent, which is almost identical to the standard drug omeprazole. The\nconsiderable decrease in the intracellular antioxidant Glutathione (NP-SH\/GSH)\nmay also be to blame for this result. Lupeol, a compound derived from the\nleaves of Ceriops decandra,<\/em> has\ndemonstrated free radical scavenging abilities that are on par with misoprostol12<\/sup>.<\/p>\n\n\n\n

As a functional, living\nfood supply for marine ecology, artemia is regarded as one of the most critical\nmarine ecotoxicity testing organisms13<\/sup>. Cytotoxicity and cell death\nare also brought on by certain polyphenols’ nonspecific attachment to the\nmembrane14,15,16,17,18,19<\/sup>.<\/p>\n\n\n\n

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

The results of our study revealed\nthat different fractions of the methanolic extract from Ceriops decandra<\/em> possess cytotoxic properties. These findings\nsuggest the need for further investigation utilizing human cell lines, both in vivo<\/em> and in vitro<\/em>. The utilization of this plant for the production of novel\nanti-cancer medications involves the identification and isolation of its\nprimary cytotoxic compound.<\/p>\n\n\n\n

Hence, more investigation is warranted to ascertain the specific phytoconstituents and underlying processes that account for the observed activities as indicated by the findings.<\/p>\n\n\n\n

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

There is no conflict of interest.<\/p>\n\n\n\n

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

This study was partly funded by Primeasia University, Banani, Dhaka and the Institute of National Analytical Research and Services (INARS), BCSIR.<\/p>\n\n\n\n

Acknowledgement<\/strong><\/p>\n\n\n\n

Authors are grateful to the Department of Pharmacy, Primeasia University, Dhaka and the Institute of National Analytical Research and Services (INARS), BCSIR.<\/p>\n\n\n\n

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

  1. Ruang-areerate, Panthita, Thippawan Yoocha, Wasitthee Kongkachana, Phakamas Phetchawang, Chatree Maknual, Wijarn Meepol, Darunee Jiumjamrassil, Wirulda Pootakham, and Sithichoke Tangphatsornruang. 2022. “Comparative Analysis and Phylogenetic Relationships of Ceriops<\/em> Species (Rhizophoraceae) and Avicennia lanata<\/em> (Acanthaceae): Insight into the Chloroplast Genome Evolution between Middle and Seaward Zones of Mangrove Forests” Biology<\/em> 11, no. 3: 383.
    CrossRef<\/a><\/li>
  2. Nabeel, Mannalamkunnath Alikunhi, Kandasamy Kathiresan, and Subramanian Manivannan. “Antidiabetic activity of the mangrove species Ceriops decandra in alloxan\u2010induced diabetic rats.” Journal of Diabetes<\/em> 2, no. 2 (2010): 97-103.
     CrossRef <\/a><\/li>
  3. Mahmud, Imran, Naznin Shahria, Sabina Yeasmin, Asif Iqbal, Emdadul Hasan Mukul, Sudipta Gain, Jamil Ahmad Shilpi, and Md. Khirul Islam. \u201cEthnomedicinal, Phytochemical and Pharmacological Profile of a Mangrove Plant Ceriops Decandra Griffdin Hou.\u201d Journal of Complementary and Integrative Medicine<\/em> 16, no. 1 (2018). https:\/\/doi.org\/10.1515\/jcim-2017-0129.
    CrossRef<\/a><\/li>
  4. John A Crairns. \u201cThe coxibs and traditional nonsteroidal anti-inflammatory drugs: A current perspective on cardiovascular risks.\u201d Canadian Journal of Cardiology <\/em>13, no. 2 (2007). https:\/\/www.onlinecjc.ca\/article\/S0828-282X(07)70732-8\/pdf <\/li>
  5. Mostofa, R., Ahmed, S., Begum, M.M. et al.<\/em> \u201cEvaluation of anti-inflammatory and gastric anti-ulcer activity of Phyllanthus niruri<\/em> L. (Euphorbiaceae) leaves in experimental rats.\u201d BMC Complement Altern Med<\/em> 17, 267 (2017). https:\/\/doi.org\/10.1186\/s12906-017-1771-7
    CrossRef <\/a><\/li>
  6. Akter, Saleha, Taslima Begum, Rayhana Begum, Tamanna Sharmin Tonny, Mousumi Yasmin, S. Shifa, and F. Afroze. “Phytochemical analysis and investigation of anti-inflammatory and anti-ulcer activity of Terminalia bellirica leaves extract.” Int J Pharmacogn<\/em> 6 (2019): 54-65<\/li>
  7. Gandagule, Upendra B., B. Duraiswamy, Mayur R. Bhurat, and Sanjay A. Nagdev. “Isolation and Characterization of Lupeol a Triterpenoid from Stem Bark of Ziziphus xylopyrus (Retz) Willd.” Inventi rapid: Pharm Analysis & Quality Assurance<\/em> 4 (2018): 1\u01e67. https:\/\/doi.org\/10.1021\/acsomega.1c05647
    CrossRef <\/a><\/li>
  8. Fatema Tuz-Zohora, Md. Abdul Muhit, Hasan CM, Ahsan M. Quinolone Alkaloids Along with Other Constituents from Zanthoxylum rhetsa and their Chemotaxonomic Significance. Records of Natural Products. 2018;12(6):634-637. doi:https:\/\/doi.org\/10.25135\/rnp.57.17.11.182
     CrossRef <\/a><\/li>
  9. Rayhana Begum, Manjur Ali Sheliya, Showkat R. Mir, Ekta Singh, Manju Sharma, \u201cInhibition of proinflammatory mediators by coumaroyl lupendioic acid, a new lupane-type triterpene from Careya arborea<\/em>, on inflammation-induced animal model\u201d, Journal of Ethnopharmacology, 206, 376-392 (2017) https:\/\/doi.org\/10.1016\/j. jep.2017.05.014.
     CrossRef <\/a><\/li>
  10. Sumsunnahar Shifa, Taslima Begum, Faruquzzaman, Farzana Afroze and Mst Kaisarunnahar Shraboni. \u201cAntibacterial activity and cytotoxic activity of leaves extract of Carissa carandas Linn.\u201d Journal of Pharmacognosy and Phytochemistry<\/em> 8, 4 (2019). https:\/\/www. researchgate.net\/publication\/361475887_Preliminary_phytochemical_screening_antibacterial_activity_and_cytotoxic_activity_of_leaves_extract_of_Carissa_carandas_Linn <\/li>
  11. Ayertey, Frederick, Ebenezer Ofori-Attah, Stephen Antwi, Michael Amoa-Bosompem, Georgina Djameh, Nathaniel Lartey Lartey, Mistuko Ohashi et al. “Anti-inflammatory activity and mechanism of action of ethanolic leaf extract of Morinda lucida Benth.” Journal of traditional and complementary medicine<\/em> 11, no. 3 (2021): 249-258. https:\/\/doi.org\/10.1016\/j.jtcme.2020.07.001
    CrossRef <\/a><\/li>
  12.  Begum, Rayhana, Manju Sharma, K. K. Pillai, Vidhu Aeri, and Manjur Ali Sheliya. “Inhibitory effect of Careya arborea on inflammatory biomarkers in carrageenan-induced inflammation.” Pharmaceutical biology<\/em> 53, no. 3 (2015): 437-445.
    CrossRef <\/a><\/li>
  13. Sahoo, Saroj Kumar, Himanshu Bhusan Sahoo, D. Priyadarshini, G. Soundarya, Ch Kishore Kumar, and K. Usha Rani. “Antiulcer activity of ethanolic extract of Salvadora indica (W.) leaves on albino rats.” Journal of clinical and diagnostic research: JCDR<\/em> 10, no. 9 (2016): FF07. doi: 10.7860\/JCDR\/2016\/20384.8470
    CrossRef <\/a><\/li>
  14. de S Lira, Silv\u00e9ria Regina, Vietla Satyanarayana Rao, Ana Carla S. Carvalho, Marjorie M. Guedes, Talita C. de Morais, Antonia L. de Souza, Maria Teresa S. Trevisan, Alana F. Lima, Mariana H. Chaves, and Fl\u00e1via A. Santos. “Gastroprotective effect of lupeol on ethanol-induced gastric damage and the underlying mechanism.”  Inflammopharmacology<\/em> 17, no. 4 (2009): 221-228.
     CrossRef <\/a><\/li>
  15. Arulvasu, Chinnasamy, Samou Michael Jennifer, Durai Prabhu, and Devakumar Chandhirasekar. “Toxicity effect of silver nanoparticles in brine shrimp Artemia.” The Scientific World Journal<\/em> 2014 (2014)x.
    CrossRef <\/a><\/li>
  16. Lim DY, Jeong Y, Tyner AL, Park JH. Induction of cell cycle arrest and apoptosis in HT-29 human colon cancer cells by the dietary compound luteolin. Am J Physiol Gastrointest Liver Physiol. <\/em>2007;292:G66\u2013G75. doi: 10.1152\/ajpgi.00248.2006
     CrossRef <\/a><\/li>
  17. Podolak I, Galanty A, Sobolewska D. Saponins as cytotoxic agents: a review. Phytochem Rev. <\/em>2010;9:425\u2013474. doi: 10.1007\/s11101-010-9183-z.
     CrossRef <\/a><\/li>
  18. Dey P, Kundu A, Chakraborty HJ, Kar B, Choi WS, Lee BM, et al. Therapeutic value of steroidal alkaloids in cancer: current trends and future perspectives. Int J Cancer. <\/em>2019;145:1731\u20131744. doi: 10.1002\/ijc.31965.
     CrossRef <\/a><\/li>
  19. Hossain SJ, Tsujiyama I, Takasugi M, Islam MA, Biswas RS, Aoshima H. Total phenolic content, antioxidative, anti-amylase, anti-glucosidase, and antihistamine release activities of Bangladeshi fruits. Food Sci Technol Res. <\/em>2008;14:261\u2013268. doi: 10.3136\/fstr.14.261.
    CrossRef <\/a><\/li><\/ol>\n","protected":false},"excerpt":{"rendered":"

    Introduction Only 124 nations and regions have at least one […]<\/p>\n","protected":false},"author":15,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[109],"tags":[],"class_list":["post-51964","post","type-post","status-publish","format-standard","hentry","category-vol16no3"],"_links":{"self":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/51964","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=51964"}],"version-history":[{"count":5,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/51964\/revisions"}],"predecessor-version":[{"id":52656,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/51964\/revisions\/52656"}],"wp:attachment":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/media?parent=51964"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/categories?post=51964"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/tags?post=51964"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}