{"id":63680,"date":"2025-03-31T10:36:24","date_gmt":"2025-03-31T10:36:24","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=63680"},"modified":"2025-04-23T03:59:48","modified_gmt":"2025-04-23T03:59:48","slug":"anti-inflammatory-and-antioxidant-potential-of-ageratum-conyzoides-ethanol-extract-against-carbon-tetrachloride-induced-renal-toxicity-in-rats","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol18no1\/anti-inflammatory-and-antioxidant-potential-of-ageratum-conyzoides-ethanol-extract-against-carbon-tetrachloride-induced-renal-toxicity-in-rats\/","title":{"rendered":"Anti-Inflammatory and Antioxidant Potential of Ageratum conyzoides Ethanol Extract Against Carbon Tetrachloride-Induced Renal Toxicity in Rats"},"content":{"rendered":"

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

The kidney is the main metabolic organ responsible for removing waste products, undesirable substances, and potentially dangerous substances from the human system1<\/sup>.<\/p>\n

Its high blood circulation and intricate cellular transport networks cause these compounds to accumulate inside nephron epithelial cells, making them highly vulnerable to the harmful effects of drugs and environmental pollutants2<\/sup>.It is necessary for animal tissues to constantly cope and deal with highly reactive oxygen species (ROS), which are generated during various metabolic physiological processes. Examples of these radicals include hydrogen peroxide, hydroxyl radicals, superoxide anion, and others. It seems that life requires some free radical generation, however, the overproduction of free radicals could lead to oxidative stress. According to a study3<\/sup>, oxidative pathways associated with the cytochrome P450 enzyme system, which are involved in the hepatic metabolism of pharmaceuticals, are responsible for almost half of all cases of acute liver failure in Ghana. In several sectors, carbon tetrachloride (CCl4<\/sub>) is a common chemical solvent. Hira and 4<\/sup> reported that kidney damage brought on by CCl4<\/sub> is similar to damage brought on by other common causes of renal deterioration, including alcohol, chemicals, viruses, and autoimmune illnesses. Kidney disease is a very prevalent and burdensome condition, and plant-based treatments have shown to be promising. It has been discovered that many plants and their extracts possess antioxidant properties, which can help mitigate kidney damage by preventing processes like inflammation, fibrosis, and oxidative stress from occurring4<\/sup>.<\/p>\n

Ageratum conyzoides<\/em> (AC), is a persistent weed that is widespread in tropical and subtropical areas5<\/sup>. Despite its reputation as a weed, the nutritional and medicinal potential of A. conyzoides<\/em> remains largely unexplored6<\/sup>. Analgesic, hemostatic, insecticidal, and antimicrobial properties have all been demonstrated for this Asteraceae family herbal plant7<\/sup>. In this study, we investigated the anti-inflammatory and antioxidant activities of Ageratum conyzoides<\/em>ethanoic crude extract against carbon tetrachloride using a rat model.\u00a0<\/strong><\/p>\n

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

Plant Collection and authentication <\/strong><\/p>\n

The Northern Region of Ghana’s Nyankpala Reserve Forest was the source of the hand-picked A. conyzoides<\/em> leaves. The leaves were verified by a and a voucher specimen (UDS\/ HM57\/ 2023 \/L011) was deposited at the herbarium. They were properly cleaned with water three times, dry at room temperature, and ground to powdered form.<\/p>\n

Preparation of extracts<\/strong><\/p>\n

One hundred grams of powdered sample of A. conyzoides<\/em> leaves were macerated using 1000 ml of 99.99 % hydroethanolic . For 24 hours at 25\u00b0C, it was shaken continuously with a shaker (Gallenkamp, England). Centrifugation was employed to process the supernatant for 20 minutes at 1106 x g at ambient temperature. Following centrifugation, the supernatants were combined and concentrated using a rotary evaporator.<\/p>\n

Preliminary Phytochemical analysis<\/strong><\/p>\n

The phytochemical components of A. conzyoides<\/em> leaves powder and ethanolic crude extracts () were identified by applying conventional techniques, as stated by Donkor and colleagues8<\/sup>. The phytochemicals that were investigated included triterpenes, alkaloids, flavonoids, saponins, polyphenols, and tannins.<\/p>\n

Evaluation <\/strong><\/p>\n

Forty-five (45) male rats, weighing between 120 and 150 g, were split up into 9 groups and treated for 7 days (n = 5). Table 1 shows the groups and treatments. The rats, which were acquired from the Noguchi Memorial Institute for Medical Research’s Animal Experimentation Unit, were housed at room temperature (25 \u00b11\u00b0C) and humidity (50\u201360%) for five days before the experiment. They also had a 12-hour light\/day cycle maintained. Agricare, Kumasi, Ghana, provided them with a standard laboratory rat diet, and they had unrestricted access to clean . The study was approved by the University for Development Studies Institutional Review Board (UDSIRB) with an ethically approved number of UDSIRB\/188\/23.<\/p>\n

The animals were given 50 mg\/kg intraperitoneal pentobarbitone to induce anesthesia on the eighth day. Through cardiac puncture, blood was obtained into EDTA (ethylenediaminetetraacetic acid)-2K tubes for hematological examination. For biochemical analyses, portions of the blood were obtained into gel-separator tubes. Afterward, the rats were euthanized, and the kidneys were removed for examination by histopathologists. Antioxidant enzymes, oxidative stress indicators, and pro-inflammatory cytokines were all measured in kidney homogenates. The animals’ carcasses were interred in a specific location following institutional ethical standards in the area.<\/p>\n

Table 1: Animal Groups and Treatments<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
S\/N.<\/strong><\/td>\nGroup<\/strong><\/td>\nTreatment<\/strong><\/td>\n<\/tr>\n
1<\/td>\nNormal (control)<\/td>\nPotable water p.o (1 ml\/kg b.wt)<\/td>\n<\/tr>\n
Extracts Only<\/td>\n<\/tr>\n
2<\/td>\n100 mg ESE only<\/td>\n100 mg\/kg ESE only (per day, p.o.) for 7 consecutive days<\/td>\n<\/tr>\n
3<\/td>\n250 mg ESE only<\/td>\n250 mg\/kg ESE only (per day, p.o.) for 7 consecutive days<\/td>\n<\/tr>\n
4<\/td>\n500 mg ESE only<\/td>\n500 mg\/kg ESE only (per day, p.o.) only for 7 consecutive days<\/td>\n<\/tr>\n
Toxicant Induced<\/td>\n<\/tr>\n
5<\/td>\nCCl4<\/sub><\/td>\nCCl4<\/sub> i.p (1 ml\/kg b.wt, 1:1 v\/v olive oil) received on 2nd<\/sup> and 3rd<\/sup> day.<\/td>\n<\/tr>\n
6<\/td>\nCCl4<\/sub> +Sily<\/td>\nSilymarin p.o (100 mg\/kg per day) received for seven days plus a single dose of CCl4<\/sub> in olive oil (1:1 v\/v, 1.0 ml\/kg, i.p.) on the 2nd<\/sup> and 3rd<\/sup> day<\/td>\n<\/tr>\n
7<\/td>\n100 mg ESE + CCl4<\/sub><\/td>\n100 mg\/kg ESE (per day, p.o.) for seven days plus a single dose of CCl4<\/sub> in olive oil (1:0 v\/v, 1.0 ml\/kg, i.p.) on the 2nd<\/sup> and 3rd<\/sup> day.<\/td>\n<\/tr>\n
8<\/td>\n250 mg ESE + CCl4<\/sub><\/td>\n250 mg\/kg ESE (per day, p.o.) for seven days for seven consecutive days plus a single dose of CCl4<\/sub> in olive oil (1:0 v\/v, 1.0 ml\/kg, i.p.) on the 2nd<\/sup> and 3rd<\/sup> day.<\/td>\n<\/tr>\n
9<\/td>\n500 mg ESE + CCl4<\/sub><\/td>\n500 mg\/kg ESE (per day, p.o.) for seven days plus a single dose of CCl4<\/sub> in olive oil (1:0 v\/v, 1.0 ml\/kg, i.p.) on the 2nd<\/sup> and 3rd<\/sup> day.<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Change in body weight of rats<\/strong><\/p>\n

The expression below was used to determine the body weights of rats after treatment:<\/p>\n

\"\"<\/p>\n

Effect of extracts on rat kidney<\/strong><\/p>\n

The absolute organ weight (AOW) of the rat\u2019s kidney was calculated and used to determine the Relative Organ Weight (ROW) as shown below:\u00a0<\/strong><\/p>\n

\"\"<\/p>\n

Hematological and Biochemical analyses<\/strong><\/p>\n

The animals’ hematological profiles were measured using an automated Sysmex XS-1000ianalyzer. Serum was extracted by centrifuging blood samples in activated gel tubes for 15 minutes at 1500x g after they had been allowed to clot. A completely automated Flexor E chemistry analyzer (VitalScientific, Japan) was used to carry out the biochemical analysis. Urea and creatinine were the twobiochemical markers that were examined. The following approach was used to calculate thepercentage of nephroprotection utilizing the primary kidney functional indicators, such as relativekidney weight, urea, and creatinine.\u00a0<\/strong><\/p>\n

\"\"<\/p>\n

Antioxidant and oxidative stress biomarkers<\/strong><\/p>\n

After homogenizing the liver (1.0 g) in 10 mL phosphate buffer (0.1 M, pH 7.4) and centrifuging at 10,000 rpm for 20 min at 4\u00b0C, the post-mitochondrial fraction was extracted (PMF). The supernatant was collected and utilized for antioxidant assays such as reduced glutathione (GSH), catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA), and myeloperoxidase (MPO) using methods as stated by Sarfo-Antwi and colleagues6<\/sup>.<\/p>\n

Inflammatory cytokine assay<\/strong><\/p>\n

Using homogenized liver samples, measurement of anti-inflammatory proteins of specific cytokines, namely Tumour Necrosis Factor-alpha (TNF-\u03b1), Transforming Growth Factor-Beta1 (TGF-\u03b21), Nuclear Factor Kappa B (NFk-\u03b2), and Cyclo-oxygenase 2 (COX-2), were tested using appropriate ELISA kits. TNF-\u03b1, NFk\u03b2, and COX-2 ELISA kits were obtained from RayBio \u2013 tech, Inc. (Norcross, GA, USA) and TGF-\u03b21 kits were from Aviscera Bioscience, Inc. (Santa Clara, CA, USA).\u00a0<\/strong><\/p>\n

Histopathological studies<\/strong><\/p>\n

The kidney sections were embedded in paraffin, fixed in 10% buffered formalin, and then cut into 4 m sections from each block. For histological analysis, the paraffin-embedded slices were stained with hematoxylin and eosin. Light microscopy (Olympus Manual System Microscope BX43) was used to conduct blinded examinations of each sample from each group while being seen at 100x and 400x magnification.<\/p>\n

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

Multiple comparison tests using Tukey were performed after a one-way ANOVA with GraphPad Prism. Statistical significance was established at the 5% level (p < 0.05), and the results were displayed as mean \u00b1 SEM.<\/p>\n

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

Preliminary Phytochemical screening<\/strong><\/p>\n

Table 2 below shows the phytochemicals present in the extract and raw powder. Alkaloids and tannins were the most abundant phytochemicals present in the extract.<\/p>\n

Table 2: Phytochemical composition of raw powder and ESE<\/strong><\/p>\n\n\n\n\n
Phytochemicals<\/strong><\/td>\nESE<\/strong><\/td>\nRaw Powder<\/strong><\/td>\n<\/tr>\n
AlkaloidsPhenolsFlavonoids<\/p>\n

Triterpene<\/p>\n

Tannins<\/p>\n

Saponins<\/td>\n

+<\/strong>+<\/strong>+<\/strong><\/p>\n

+<\/strong><\/p>\n

+<\/strong><\/p>\n

+<\/strong><\/td>\n

+<\/strong>+<\/strong>+<\/strong><\/p>\n

+<\/strong><\/p>\n

+<\/strong><\/p>\n

+<\/strong><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Key: Present (+); Absent (-).\u00a0<\/strong><\/p>\n

Body weight after Treatment<\/strong><\/p>\n

Rats treated with CCl4<\/sub>-treated only, observed a substantive reduction in weight as shown in Table 3.<\/p>\n

Table 3: Variation in Weight of Rats<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Treatment<\/strong><\/td>\n% Change in Body Weight, D7<\/strong><\/td>\n<\/tr>\n
Normal<\/td>\n3.11 \u00b1 1.02b<\/sup><\/td>\n<\/tr>\n
Extracts only<\/em><\/strong><\/td>\n<\/td>\n<\/tr>\n
100 mg\/kg b.wt. ESE only<\/td>\n1.18 \u00b1 0.12a<\/sup><\/td>\n<\/tr>\n
250 mg\/kg b.wt. ESE only<\/td>\n2.35\u00b1 0.25b<\/sup><\/td>\n<\/tr>\n
500 mg\/kg b.wt. ESE only<\/td>\n3.23 \u00b1 1.31b<\/sup><\/td>\n<\/tr>\n
CCl4<\/sub>only<\/td>\n-2.32 \u00b1 1.04a<\/sup><\/td>\n<\/tr>\n
Sily+CCl4<\/sub><\/td>\n2.75 \u00b1 0.15b<\/sup><\/td>\n<\/tr>\n
100 mg\/kg b.wt. ESE +CCl4<\/sub><\/td>\n0.21 \u00b1 0.47ab<\/sup><\/td>\n<\/tr>\n
250 mg\/kg b.wt. ESE +CCl4<\/sub><\/td>\n0.57 \u00b1 0.05ab<\/sup><\/td>\n<\/tr>\n
500 mg\/kg b.wt. ESE +CCl4<\/sub><\/td>\n1.25 \u00b1 0.11ab<\/sup><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Mean \u00b1 SEM (n=5) is used to express values. The significance level is “a” at p<0.05-0.001 relative to the normal group and “b” at p<0.05-0.001 when relative to the CCl4<\/sub>-treated group alone.<\/p>\n

Relative Weight of Organs Following Therapy<\/strong><\/p>\n

Figure 1 illustrates that in comparison to the normal group, the relative organ weight of animals treated with CCl4<\/sub> only increased significantly (p 0.05).<\/p>\n\n\n\n
\"\"<\/td>\nFigure 1: Treatment’s Impact on Relative Organ Weight. The results are presented as mean \u00b1 SEM (n=5). Statistical significance “a” between 0.05 and 0.001 in comparison to Normal; “b” between 0.05 and 0.001 in comparison to the CCl4<\/sub>-treated group only.<\/p>\n

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

The impact of therapy on hematological parameters<\/strong><\/p>\n

The hematological indices were measured to assess the effect of the CCl4<\/sub>, silymarin, and ESE on the rats. In general, therapy had no significant effect on the rats’ hematological indicators, as indicated in Table 4.<\/p>\n

Table 4: Treatment’s Impact on Haematological Markers<\/strong><\/p>\n\n\n\n\n\n\n\n
Parameter<\/td>\nHGB(g\/dL)<\/td>\nWBC(103<\/sup>\/\u00b5L)<\/td>\nRBC(106<\/sup>\/\u00b5L)<\/td>\nPLT(103<\/sup>\/\u00b5L)<\/td>\n<\/tr>\n
Normal<\/td>\n11.43 \u00b1 1.32<\/td>\n7.13 \u00b1 1.21<\/td>\n5.83 \u00b1 2.20<\/td>\n713.34 \u00b1 32.12<\/td>\n<\/tr>\n
Extract Only<\/em><\/td>\n<\/tr>\n
100 mg\/kg b.wt. ESE only250 mg\/kg b.wt. ESE only500 mg\/kg b.wt. ESE only<\/td>\n12.11 \u00b1 2.3411.34 \u00b1 1.5211.10 \u00b1 1.61<\/td>\n9.51 \u00b1 2.3212.21 \u00b1 1.268.30 \u00b1 1.42<\/td>\n5.62 \u00b1 1.315.22 \u00b11.125.95 \u00b1 1.45<\/td>\n764.85 \u00b1 43.65843.10 \u00b1 38.47821.83 \u00b1 54.23<\/td>\n<\/tr>\n
CCl4<\/sub> onlySilymarin + CCl4<\/sub>100 mg\/kgb.wt.ESE+ CCl4<\/sub><\/p>\n

250 mg\/kgb.wt.ESE+ CCl4<\/sub><\/p>\n

500 mg\/kgb.wt.ESE + CCl4<\/sub><\/td>\n

7.11 \u00b1 1.43a<\/sup>11.07 \u00b1 1.33b<\/sup>9.43 \u00b1 1.42ab<\/sup><\/p>\n

9.51 \u00b1 0.85ab<\/sup><\/p>\n

10.86 \u00b1 1.19b<\/sup><\/td>\n

5.02 \u00b1 1.17a<\/sup>8.33 \u00b1 1.25ab<\/sup>6.76 \u00b1 2.12<\/p>\n

6.26 \u00b1 1.64<\/p>\n

7.42 \u00b1 1.32b<\/sup><\/td>\n

3.02 \u00b1 0.35a<\/sup>3.61 \u00b1 1.24a<\/sup>3.98 \u00b1 1.11a<\/sup><\/p>\n

3.82 \u00b1 1.32a<\/sup><\/p>\n

4.17 \u00b1 1.73<\/td>\n

1165.54 \u00b1 70.23a<\/sup>965.12 \u00b1 36.16ab<\/sup>1012.43 \u00b1 50.87a<\/sup><\/p>\n

1000.97 \u00b1 67.82a<\/sup><\/p>\n

865.74 \u00b1 43.73b<\/sup><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

The data are shown as mean\u00b1SEM (n=5). Significant differences between the Normal and CCl4<\/sub>-treated groups were statistically indicated by “a” at p<0.05-0.001 and “b” at p<0.05-0.001 respectively.<\/p>\n

The impact of remedies on serum urea and creatinine<\/strong><\/p>\n

Figure 2 shows that rats administered CCl4<\/sub> alone had substantially higher creatinine and urea levels than the normal group.<\/p>\n\n\n\n
\"\"<\/td>\nFigure 2:\u00a0 Effects of treatments on serum creatinine and urea. The findings (n=5) are shown as mean \u00b1 SEM. Statistical significance: “a” compares to Normal at p<0.05-0.001, and “b” compares to the group that received only CCl4<\/sub> at p<0.05-0.001.<\/strong><\/p>\n

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

Impact of therapy on percentage protection<\/strong><\/p>\n

Figure 3 depicts the kidney-protective impact of Silymarin and ESE at 100, 250, and 500 mg\/kg b.wt. against CCl4<\/sub>induction. ESE at 500 mg\/kg b.wt provided the most effective therapy against CCl4 at 96%, surpassing standard silymarin at 93%.<\/p>\n\n\n\n
\"\"<\/td>\nFigure 3: shows the percentage of protection provided by silymarin and all doses of ESE against carbon tetrachloride. The individual bar represents the average percentage protection of the kidney biomarkers.<\/strong><\/p>\n

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

Treatments’ effects on antioxidant indicators<\/strong><\/p>\n

Table 4 presents the effects of therapy on oxidative stressors and renal antioxidants. When comparing the CCl4<\/sub>-treated group to the Normal group, there was a notable rise in the oxidative stress biomarker (MDA) and a substantial decrease in the antioxidant assay (CAT, SOD, and GSH) following CCl4<\/sub> administration. Comparing the CCl4<\/sub> group to the co-treated groups with ESE at all dosages, however, revealed substantial (p<0.05) differences in antioxidant and oxidative stress biomarker levels.<\/p>\n

Table 5: Effect of Treatment on Antioxidant Makers<\/strong><\/p>\n\n\n\n\n\n\n\n
Parameter<\/strong><\/td>\nGSH<\/strong>(\u03bcmol\/mg<\/strong>protein)<\/strong><\/td>\nCAT<\/strong>(U\/mg Protein)<\/strong><\/td>\nSOD<\/strong>(U\/mg Protein)<\/strong><\/td>\nMDA<\/strong>(mmol\/mg<\/strong>protein)<\/strong><\/td>\n<\/tr>\n
Normal<\/td>\n187.75 \u00b1 9.23<\/td>\n50.42 \u00b1 8.12<\/td>\n3.12 \u00b1 0.75<\/td>\n10.83 \u00b1 2.98<\/td>\n<\/tr>\n
Treatment (without <\/em>CCl4<\/sub>)<\/em><\/td>\n<\/tr>\n
100 mg\/kg b.wt. ESE only250 mg\/kg b.wt. ESE only500 mg\/kg b.wt. ESE only<\/td>\n180.76 \u00b1 4.34192.18 \u00b1 8.54188.23 \u00b1 6.07<\/td>\n54.24 \u00b1 9.1152.71 \u00b1 4.2655.42 \u00b1 7.93<\/td>\n2.98 \u00b1 0.433.40 \u00b11.053.21 \u00b1 1.11<\/td>\n8.34 \u00b1 2.419.45 \u00b1 1.487.98 \u00b1 1.23<\/td>\n<\/tr>\n
CCl4<\/sub>onlySilymarin + CCl4<\/sub>100 mg\/kg b.wt. ESE + CCl4<\/sub><\/p>\n

250 mg\/kg b.wt. ESE+ CCl4<\/sub><\/p>\n

500 mg\/kg b.wt. ESE+ CCl4<\/sub><\/td>\n

84.73 \u00b1 3.54a<\/sup>172.41 \u00b1 5.33b<\/sup>102.06 \u00b1 6.87ab<\/sup><\/p>\n

131.52 \u00b1 4.82ab<\/sup><\/p>\n

162.75 \u00b1 8.22ab<\/sup><\/td>\n

14.39 \u00b1 5.36a<\/sup>30.58 \u00b1 6.04ab<\/sup>26.92 \u00b1 3.53ab<\/sup><\/p>\n

33.17 \u00b1 4.02ab<\/sup><\/p>\n

46.11 \u00b1 5.32b<\/sup><\/td>\n

0.83 \u00b1 0.10a<\/sup>3.21 \u00b1 0.35b<\/sup>2.04 \u00b1 0.36ab<\/sup><\/p>\n

2.75 \u00b1 0.66b<\/sup><\/p>\n

3.54 \u00b1 0.73b<\/sup><\/td>\n

98.63 \u00b1 7.62a<\/sup>20.11 \u00b1 6.16ab<\/sup>46.42 \u00b1 8.25ab<\/sup><\/p>\n

35.19 \u00b1 5.32ab<\/sup><\/p>\n

26.24 \u00b1 5.03ab<\/sup><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

The results are shown as mean \u00b1 SEM (n=5).\u00a0 “a” represents the statistical significance when compared to the Normal group, and “b” represents the statistical significance when compared to the CCl4<\/sub>-treated group only.<\/p>\n

Treatment impact on inflammatory markers<\/strong><\/p>\n

Inflammatory markers were substantially higher in the group treated with only CCl4<\/sub> than in the normal group (p<0.05-0.001).<\/p>\n\n\n\n
\"\"<\/td>\nFigure 4: depicts the impact of therapy on inflammatory biomarkers. Data are presented as mean \u00b1 SEM (n=5). “a” indicates a significant distinction from the normal group (P < 0.05 – 0.001), while “b” indicates a significant variance from the CCl4<\/sub>-treated group.\u00a0<\/strong><\/p>\n

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

Impact of Therapy on Liver Histology<\/strong><\/p>\n

The kidneys of the normal group recorded no undesired morphological changes; nevertheless, the group receiving CCl4<\/sub>only had significant diffuse tubular ectasia with coagulation necrosis. Animals who received 500 mg\/kg bwt of ESE recovered almost normal architecture in place of these abnormalities as shown in Figure 4.<\/p>\n\n\n\n
\"\"<\/td>\nFigure 5: depicts the impact of therapy on kidney histology. (a) – (d) show no visible damage; (e) indicate diffuse tubular ectasia with epithelial coagulation necrosis; (f) – (h) show mild coagulation necrosis; (j) display no observable lesion.\u00a0<\/strong><\/p>\n

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

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

Numerous articles have shown that a variety of mutagens and carcinogens can create peroxide radicals which can initiate the formation of cancer and other health disorders9-12<\/sup>.<\/p>\n

The current study sought to assess any potential safeguards provided by ESE against CCl4<\/sub>-induced oxidative stress, which causes damage to the kidneys in rats. Phytochemical analysis of ESE and raw powder revealed the existence of alkaloids, phenols, flavonoids, triterpenes, tannins, as well as saponins. CCl4<\/sub> is a widely used, trusted, and well-known chemical that can harm the liver and kidney tissue13-15<\/sup>. As expected, the CCl4<\/sub>-treated group had a significant reduction in body weight at termination because it interfered with the animal’s normal metabolism of fat, protein, and carbohydrates. Additionally, a decrease in appetite directly affects body weight reduction. However, co-treatment with ESE at all dosages resulted in a significant increase in body weight. Figure 1 illustrates how co-treatment with extracts returned kidney weight to a nearly normal range, despite the possibility that the negative effects of CCl4<\/sub> caused the rats’ kidneys to enlarge and allow materials like fatty acids and glycerol to enter the kidney cells. According to past research, CCl4<\/sub> treatment increased several organs, including the kidney and liver16-18<\/sup>.<\/p>\n

The current investigation found that serum creatinine and urea levels were considerably higher in rats who received CCl4<\/sub> induction as shown in Figure 2. Lipid peroxidation caused a loss in membrane integrity and the accumulation of indicators like creatinine and urea in damaged kidneys brought on by CCl4<\/sub> intoxication. However, pre-treatment with silymarin and ESE at all doses resulted in a significant reduction in these markers.<\/p>\n

This was confirmed by the histological observation, which revealed widespread tubular ectasia in the CCl4<\/sub> alone group together with coagulation necrosis of epithelium and kidney interstitial inflammation as shown in Figure 5. As seen in Figure 3, these results indicate that ESE and silymarin can protect nephrocytes from CCl4<\/sub>-induced injury. The maximum protection effect of the kidney was achieved with pretreatment ESE at 500 mg\/kg b.wt. Erythropoietin synthesis by the kidney is disrupted by CCl4<\/sub>, which also alters erythrocyte viability and morphology19<\/sup>.In this study, the reduction of hemoglobin synthesis and the reduction of erythrocyte life may have contributed to the anemia associated with CCl4 <\/sub>as seen in Table 4. Similar findings postulated that CCl4<\/sub> direct detrimental effects on the cell membrane were the reason for the rats’ shortened life span20-23<\/sup>.<\/p>\n

Additionally, the capacity of iron to generate hemoglobin in the mitochondria is inhibited by CCl4<\/sub>24-25<\/sup>. Free radical processes are known to be involved in the kidney damage caused by CCl4<\/sub>25<\/sup>. A significant rise in MDA levels and a significant reduction in antioxidant makers were seen in rats exposed to CCl4<\/sub>, however, as Table 5 illustrates, rats who received both CCl4<\/sub> and ESE concurrently showed a considerable drop in MDA and an increase in antioxidant enzymes. Histological studies have demonstrated that low levels of antioxidant enzymes could cause oxidative stress and lipid peroxidation. In this study, pre-treatment with ESE at all dosages restored normal levels of TNF-\u03b1, TGF-\u03b21, NF \u043a\u03b21, and COX-2, which were significantly elevated owing to CCl4<\/sub> toxicity. These outcomes align with a previous investigation on CCl4<\/sub> poisoning, whereby molecular discoveries demonstrated increased quantities of TNF-\u03b1, TGF-\u03b21, COX-2, IL-17, and IL-23, in addition to overexpression of NF \u043a\u03b2 in the hepatic tissues26<\/sup>.<\/p>\n

-inflammatory cytokines including TNF-\u03b1, TGF-\u03b21, COX-2, IL-17, and IL-23 are produced through the mitogen-activated protein kinase (MAPK) pathway, which is reported to be activated by CCl4<\/sub>27-29<\/sup>.Furthermore, CCl4<\/sub> participates in the activation of NF-\u043a\u03b2, which regulates the production of several genes linked to inflammation, including adhesion molecules, chemokines, and pro-inflammatory cytokines30<\/sup>.Researchers in this study discovered that lowering the amounts of pro-inflammatory cytokines in kidney homogenates after ESE treatment at all dosages also reduced inflammatory responses. The anti-inflammatory mechanism of ESE therapies is suggested by their reported down-regulation of NF-\u03ba\u03b2 expression along with associated cytokines and chemokines, hence providing a nephroprotective effect. The protective impact of kidneys through inflammation suppression is indicated by positive regulation of TNF-\u03b1 production by ESE. Nephrocytes are known to express cyclooxygenase 2 (COX-2), which both attenuates and stimulates cell proliferation in acute kidney damage and only occurs under conditions of prolonged aggressiveness. Furthermore, inflammatory neurotoxicity has been linked to COX-2 activation after lead exposure31-32<\/sup>.<\/p>\n

According to the current study, CCl4<\/sub> causes over-production of COX-2 in kidney tissues, which in turn causes excessive inflammation (together with NF-\u03ba\u03b2 and TNF-\u03b1). However, COX-2 expression was downregulated when co-treated with ESE and the standard drug, silymarin. This provides more proof of the extract’s anti-proliferative ability in nephroprotection. The study’s findings demonstrated that ESE had a protective effect at all dosages, with the highest protection occurring at 500 mg\/kg b.wt. The biochemical and histological data suggest that ESE is nephroprotective, which is likely due to the phytochemicals present.<\/p>\n

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

Our findings validated the antioxidant characteristics of ESE and showed, for the first time, that these benefits are linked to kidney protection through the suppression of anti-inflammatory cytokines. These findings corroborate previous research that suggested medicinal plants were a good substitute for kidney protection. We concluded that ESE could be essential for its economic use and perhaps further study could look into separating the pure bioactive chemicals that give its medicinal effect.<\/p>\n

Acknowledgment<\/strong><\/p>\n

The authors are grateful to the University for Development Studies for providing the laboratory to support this study.<\/p>\n

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

The author(s) received no financial support for the research, authorship, and\/or publication of this article.\u00a0<\/strong><\/p>\n

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

The author(s) do not have any conflict of interest.<\/p>\n

Data Availability Statement<\/strong><\/p>\n

This statement does not apply to this article.<\/p>\n

Ethical statements<\/strong><\/p>\n

The study was approved by the University for Development Studies Institutional Review Board (UDSIRB) with an ethically approved number of UDSIRB\/188\/23<\/p>\n

Informed Consent Statement<\/strong><\/p>\n

This study did not involve human participants, and therefore, informed consent was not required.<\/p>\n

Clinical Trial Registration<\/strong><\/p>\n

This research does not involve any clinical trials<\/p>\n

Author Contributions<\/strong><\/p>\n

Conceptualization and methodology: Frederick Sarfo-Antwi, Christopher Larbie, and Abdul-Rashid Lansah Adam. Formal analysis and investigation: Frederick Sarfo-Antwi, Benjamin Obukowho Emikpe, Christopher Larbie and Abdul-Rashid Lansah Adam. Writing of the original draft, review, and editing: Frederick Sarfo-Antwi, \u00a0Rashid Lansah Adam and Jennifer Suurbaar. All authors approved the final manuscript.<\/p>\n

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