CHEMICAL CONSTITUENTS AND HEPATOPROTECTIVE EFFECT OF CASSIA OCCIDENTALIS L. CULTIVATED IN EGYPT

                                                                                                                 AssiutUniversity web-site: www.aun.edu.eg

CHEMICAL CONSTITUENTS AND HEPATOPROTECTIVE EFFECT OF CASSIA OCCIDENTALIS L. CULTIVATED IN EGYPT

HANAA M. SAYED¹; MAHMOUD A. RAMADAN¹; MANAL M. SAYED²; SARY KH. ABD ELGHAFFAR³ and HEBA H. SALEM¹

¹ Department of Pharmacognosy, Faculty of Pharmacy, AssiutUniversity, Assiut71526, Egypt.

² Department of Chemistry, Animal Health Research Institute, Assiut Lab., Egypt.

³ Department of pathology and Clinical Pathology, Faculty of Vet. Med.AssiutUniversity, Assiut71526, Egypt.

Received: 31 March 2016 ;       Accepted:  30 April 2016

INTRODUCTION

Liver diseases are considered a global problem especially in developing countries including Egypt. Hepatitis, viral infection, food additives, alcohol, toxic industrial chemical as well as air and water pollutants are the major risk factor of liver toxicity (Khan et al., 2012). Treatment of liver diseases via synthetic medication is usually disappointing due to its sever undesirable effect upon prolonged administration. Therefore, herbal products gain more interest in this issue and most recent studies aim to characterize the health promoting properties of many plants, especially those rich in phenolics that known

Corresponding author: Dr. SARY KH. ABD ELGHAFFAR

E-mail address: sary64@windowslive.com

Present address: Department of pathology and Clinical Pathology, Faculty of Vet. Med.AssiutUniversity, Assiut71526, Egypt.

to possess hepatoprotective and anti-inflammatory potencies.

Cassia is a genus of 40 species belonging to family Fabaceae, subfamily Caesalpinoideae (Tackhol, 1974). C. occidentalis L. (Coffee senra, Negro Coffee) as an Indian perennial diffuse under shrub, is used in several traditional medicine to cure various disease (Khare, 2007). This plant possess antibacterial, anthelmintic (Shao et al., 2013), antifungal, antiviral (Cong et al., 2014), antispasmodic, analgesic, antipyretic (Nakamura et al., 2014), antiplatelet aggregation, antihistamine release, hepatoprotective and neuroprotective (Kim et al., 2007 and Sung et al., 2004). Phytochemical studies of C. occidentalis reveled the presence of diversity of constituents including tannins, saponins, flavonoids, terpenes and anthraquinones (Bukhari et al., 2014 and Yadava et al., 2012).

Carbon tetrachloride (CCl₄) is one of the chemicals which cause liver damage through lipid peroxidation and oxidative stress (Moreira et al., 2014). CCl₄ is a suitable chemical to induce oxidative liver toxicity in experimental animal model and its toxic effects was extensively studied by several investigators (Al-Dbass et al., 2012 and Azeem et al., 2010). In recent literatures it is documented that natural drugs with antioxidant potential can protect the liver from damage caused by CCl₄ (Andritoiu et al., 2014; Shaker et al., 2010 and Sanmugapriya and Vankataraman 2006).

The current study was planned to evaluate the hepato and kidney protective activities of the fractionated methanolic extract of the aerial part of C. occidentalis L. as well as, to isolate and identify the components of the potent extractive.

MATERIALS AND METHODS

1- Plant Material:

Aerial parts of C. occidentalis L. were collected in June 2013 from plants cultivated in the Experimental Station of Medicinal Plants, Faculty of Pharmacy, AssiutUniversity. The plant was kindly authenticated by Prof. Dr. A.A. Fayed, Professor of Plant Taxonomy, Faculty of Science, AssiutUniversity. Voucher specimen of the plant material was deposited at the Herbarium of Pharmacognosy Department, Faculty of Pharmacy, AssiutUniversity.

2- Plant Extraction:

Air dried powdered aerial parts of C. occidentalis L. (3 kg) was extracted by percolation with 80% methanol (4x6 L). The resulting methanolic extract was evaporated under vacuum, at a temperature not exceeding 40°C. The concentrate (300 gm) was suspended in distilled water (600 ml), transferred to a separating funnel and subjected to successive several solvent fractionation using n- hexan, chloroform (CHCl₃), ethyl acetate (EtOAc) and n-butanol. The different extractives were concentrated separately under reduced pressure to give 65, 35, 25 and 8 gm, respectively. The remaining aqueous extract was kept for further study (Harborne, 1973).

3- Instruments and Chemicals:

Nuclear magnetic resonance (NMR) spectra were recorded on Bruker Spectrophotometer at 400 MHz for ¹H and 100 MHz for ¹³C NMR. Digital VIS/Ultraviolet Spectrophotometer was recorded on Cecil, Cambridge, England. Olympus microscope CX31, Japan and Camera (Olympus, Camedia, C-5060, Japan) were also used.

All chemicals and solvents were of analytical grade. Silica gel G₆₀ (Prolabo, India) for column chromatography, precoated silver gel plats G₆₀ F₂₅₄ (Aluminum sheets, E-Merck, Germany), for thin layer chromatography (TLC). The biochemical kits were purchased from Bio-diagnostics, Cairo, Egypt.

4- Experimental Animals:

Sixty-six male Wistar albino rats (120-150 g) were obtained from the Animal House, Faculty of Medicine, AssiutUniversity. All animals were acclimatized to the laboratory conditions, kept in controlled environment of air and temperature for 15 days before starting the experiments with access of water and diet ad Libitum.

5- Acute Toxicity Study of Plant Extract:

Three groups of rats (8 rats, each) were examined for the acute toxicity of the plant extract. One oral dose of C. occidentalis methanolic extract at concentrations of 200, 400 and 800 mg/kg B.W. were administered to rats. The mortality rate was calculated 24 hours after administration. The total methanolic extract of C. occidentalis aerial parts did not show any signs of toxicity and mortality up to 4 g/kg B.W. according to Lorke (1983).  

6- Experimental Design:

C. occidentalis different extractives (5 g, each) were suspended in Tween in distilled water at the appropriate concentration prior to oral treatment. The administration regimen was started one day after the injection of CCl₄ and extended for two weeks. Sixty-six male Wistar albino rats were divided into 11 groups (6 animals, each).

Group 1: (Control group) received Tween only.

Group 2: (CCl₄ group) injected (I.P.) with one dose of CCl₄ in olive oil (1:1) 2 mg/kg B.W. according to Lutz et al., (2003).

Group 3: (Silymarin reference group) injected (I.P.) with CCl₄ in the same dose and followed by Silymarin (25 mg/kg b.wt.) orally for 7 days according to Mohamed (2013).

Groups 4-11: Served as testing groups and injected with CCl₄ in the same previous dose and followed by oral daily administration by one of either 200 or 400 mg/kg B.w. from each of the four extractives. The treatment started one day after the injection of CCL₄ and extended for 2 weeks at the same time for the same duration.

7- Serum Sample Preparation:

Blood was collected from the retro-orbital plexus into clean free test tubes. Serum was prepared after centrifugation of the blood samples at 3000 r.p.m for 10 minutes and stored at ^_20ºC C for further estimation of liver and kidney function tests.

8- Biochemical Estimations:

Serum AST, ALT and bilirubin as well as serum urea and creatinine were measured by using the appropriate test kits (Bio- diagnostics, Cairo, Egypt) spectrophotometrically. This was done according to the standard protocol of the suppliers. 

9- Histopathological Examination:

Tissue specimens from liver were fixed in 10% neutral buffered formalin, dehydrated in a graded ethyl alcohol series, cleared with methyl benzoate and embedded in paraffin wax. Sections of 5 µm were cut and stained with haematoxylin and eosin (Bancroft et al., 1996). Stained sections were examined under light microscope and photographed using digital camera.

10- Statistical Analysis:

Data are expressed as mean ± SE for all parameters. The data were analyzed using Graph Pad Prism Data Analysis program software package (Graph Pad Software, Inc., San Diago CA, USA) for multiple comparisons, one-way analysis of variance (ONE-WAY-ANNOVA) test was used.

11- Isolation of Ethyl Acetate Extractive Contents:

Fractionation of the ethyl acetate extractive (10 g) was done on silica gel column (150x5 cm, 400 g) and eluted successively with CH₂Cl₂-methanol gradient to give several fractions screened by TLC and similar fractions were combined to afford four fractions (I-IV) (2, 1.5, 3 and 2.5 g, respectively). Fraction I (2 g) was rechromatographed on silica gel column (100x2 cm/80 g) with gradient elution using CH₂Cl₂-MeOH in 2% increments to yield compounds (1) and (2). Fraction II (1.5 g) was rechromatographed on silica gel column (100x2 cm, 60 g) with gradient elution using CH₂Cl₂-MeOH in 1% increments to yield compounds (3) and (4). Fraction III (3 g) were rechromatographed on silica gel column (100x2, 120 g), with gradient elution using CH₂Cl₂-MeOH in 1% increment to yield compound (5) and (6). Fraction IV showed mixture of minor spots, was kept for further investigation.

RESULTS

I- Biochemical Study:

Analysis of serum parameters of liver function tests (Table 1) demonstrated a decrease in serum concentration of ALT, AST and total bilirubin in groups which received CCl₄ and treated either with total methanolic extract or different extractives of C. occidentalis L. when compared with the group which received CCl₄only.

The two groups which treated with EtoAc extractive (200 & 400 mg/ kg  B.W.) showed the best improvement in liver function parameters in which there was marked significant decrease in ALT, AST and total bilirubin concentractions. Comparing this improvement in liver function parameters with those of sylimarin treated group, no significant changed could be detected. Groups which treated with 200 mg /kg  B.W. of  CHCl₃ and 200 & 400 mg/kg B.W. of n-butanol extractive showed non-significant decrease in ALT values.

Analysis of serum parameters of kidney function tests (Table 2) showed non-significant decrease in serum urea values in groups which received CCl₄ and treated with all extractives when compared with those of the group which received CCl₄ alone.

Groups which treated with EtoAc extractive (200 & 400 mg/kg B. W.) showed a significant decrease in urea concentration when compared with that of CCl₄ alone. In addition, no significant changed in urea concentration could be detected in comparison with that of Tween and Silymarin groups.

Concerning serum creatinine level in groups which treated with 200 & 400 mg/ kg B.W. of EtOAc extractive, significant decrease was detected on comparison with that of CCl₄ received group. In addition, no significant changes could be recognized in serum creatinine level when compared with those of Tween and Silymarin groups. Other groups which treated with total methanolic, CHCl₃ and n-butanol extractives showed non-significant decrease in serum creatinine level in comparison with CCl₄ received group.

Histopathological Examination:

Microscopic examination of the liver sections of negative control group (Tween) revealed normal histological appearance of the liver tissue with normal hepatolobuler architecture contained central vein and regular hepatic cell cords (Fig. 1A), while that from reference group (Silymarin plus CCl₄), showed mild vacuolar degeneration of the hepatocytes (Fig. 1B). In CCl₄ treated group the hepatocytes showed marked fatty degeneration with appearance of small and large cytoplasmic vacuoles (Fig. 1C). In group received CCl₄ along with 200 mg total methanolic extract, the liver cells showed moderate fatty degeneration (Fig. 1D), while in group treated with 400 mg total methanolic extract, the hepatocytes surrounding the central vein showed minute foci of necrosis (Fig. 2A). In the group treated with 200 mg and 400 mg CHCl₃ extractive, the hepatocytes showed very mild vacuolar degeneration (Fig. 2B&C). Liver examination of group treated with 200 mg EtOAc extractive showed normal hepatocytes with vesicular nuclei and acidophilic cytoplasm (Fig. 2D), while in 400 mg EtOAc extractive treated group the liver showed congestion of the central vein with very mild vacuolar degeneration of the hepatocytes (Fig. 3A). In n-butanol extractive 200 mg treated group, the liver showed marked vacuolar degeneration of the hepatocytes and periaciner necrobiosis (Fig. 2B), while in group received 400 mg n-butanol extractive, the liver showed congestion of the central vein with marked vacuolar degeneration of the hepatocytes (Fig. 3C).

Isolated Compounds:

Fractionation of the ethyl acetate extractive of the methanolic extract of C. occidentalis L. that showed significant hepatoprotective effect resulted in isolation of 6 phenolic compounds (Fig. 4). All of them were identified basically on chromatographic, chemical and spectroscopic analysis as well as through comparison with published data.

Compound (1) was obtained as yellow amorphous powder (20 mg, MeOH), attained deep purpule colour under UV and a yellow to orange colour with AlCl₃ spray. UV l_max nm: MeOH (287, 326), +NaOMe (245, 323), +NaOAc (284, 323), +NaOAc/H₃BO₃ (290, 390), +AlCl₃ (312, 382), +AlCl₃/HCl (305, 372). ¹H NMR (400 MHz, DMSO-d₆) d ppm: 5.42 (1H, dd, J= 14.4,2.4, H-2), 3.25 (1H, dd, J= 12.8, 3.2, H-3a), 2.70 (1H, dd, J= 12.8, 3.2, H-3b), 5.86 (2H, s, H-6,8), 6.79 (2H, d, J= 8.4, H-3',5'), 7.31 (2H, d, J= 8.4, H-2',6'), 12.14 (1H, s, 5-OH). ¹³C NMR (100 MHz, DMSO-d₆) d ppm: C-2 (78.88), C-3 (42.41), C-4 (196.81), C-5 (163-39), C-6 (96.28), C-7 (163.94), C-8 (95.46), C-9 (158.15), C-10 (102-19), C-1' (129.32), C-2',6' (128.80), C-3',5' (115.63),C-4'(167.22). The above mentioned data is in agreement with that reported for naringenin (Mabry et al., 1970 and Agrawal (1989). This is the first report for its isolation from the studied plant.

Compound (2) was obtained as yellow amorphous powders (20 mg, MeOH), attained yellow colour under UV and intensified after AlCl₃ spray. UV l_max nm: MeOH (255, 370), +NaOMe (275, 425), +NaOAc (266, 390), +NaOAc/H₃BO₃ (259, 385), +AlCl₃ (272, 425), +AlCl₃/HCl (261, 423) nm. ¹H NMR (400 MHz, DMSO-d₆) d ppm: 6.18 (1H, d, J= 2, H₆), 6.4 (1H, d, J= 2, H₈), 6.88 (1H, d, J= 8.4, H_5'), 7.53 (1H, d, J= 8.4, H_6'), 7.67 (1H, d, J= 2, H_2'). The above mentioned data is consistent with the literature data of quercetin (Mabry et al., 1970 and Agrawal 1989). It was previously isolated from the plant.

Compound (3) was obtained as yellowish orange (10 mg, MeOH). UV l_max nm: MeOH (225, 277, 429). ¹H NMR (400 MHz, DMSO-d₆) d_H ppm: 7.83 (2H, t, J= 8, H-3,-6), 7.73 (2H, d, J= 8.4, H-4,5), 7.40 (2H, d, J= 8.4, H-2,7), 11.94 (s, OH). ¹³C NMR (100 MHz, DMSO-d₆) d_c ppm: C-1,8 (161.83), C-2,7 (124.90), C-3,6 (137.93), C-4,5 (119.78), C-8a,9a (133.81), C-4a,5a (116.47), C-9 (191,62), C-10 (181.37). The above mentioned data is in agreement with that previously reported for 1,8-dihydroxy anthraquinone, which was previously isolated from the roots of the plant. (Chukwujekwu et al., 2006).

Compound (4) was obtained as yellowish orange (10 mg, MeOH). ¹H NMR (400 MHz, DMSO-d₆) d_H ppm: 7.77 (1H, s, H₂), 8.13 (1H, s, H₄), 7.75 (1H, d, J= 7.6, H₅), 7.83 (1H, t, J= 7.6, H₆), 7.41 (1H, d, J= 8, H₇), 11.98 (OH). ¹³C NMR (100 MHz, DMSO-d₆) d_c ppm: C₁-1 (161.5), C-2 (124.64), C-3 (165.92), C-4 (119.22), C-4a (134.24), C-5 (119.22), C-6 (138.13), C-7 (129.15), C-8 (161.83), C-8a (116.33), C-9 (191.62), C-9a (118.53), C-10 (181.37), C-10a (133.14). The above mentioned data is consistent with the literature data of 1,3,8-trihydroxy anthraquinone (Rogerio et al., 2008). It is the first report for its isolation from the plant.

Compound (5) was obtained as yellow amorphous powder (15 mg, MeOH) attained deep purple colour under UV and a yellow colour with AlCl₃ spray. UV l_max nm: MeOH (267, 342), +NaOMe (270, 391), +NaOAc (269, 346), +NaOAc/H₃BO₃ (269, 350), +AlCl₃ (273, 347), +AlCl₃/HCl (275, 349, 290). ¹H NMR (400 MHz, DMSO-d₆) d ppm: 6.46 (1H, s, H-6), 6.76 (1H, s, H-3), 6.82 (1H, s, H-8), 7.13 (1H, d, J= 8.4, H-5'), 7.44 (1H, s, H-2'), 7.57 (1H, d, J= 8.4, H-6'), 5.08 (1H, d, J= 7.2, H-1') 4.53 (1H, d, J= 1.2, H-1"'), 3.87 (3H, s, OCH₃), 3.17-5.45 (m, sugar protons), 1.07 (3H, d, J= 6, H-6'"), 12.93 (1H, s, 5-OH). ¹³C NMR (100 MHz, DMSO-d₆) d ppm: C-2 (163.41), C-3 (104.29), C-4 (182.42), C-5 (161.67), C-6 (100.05), C-7 (164-66), C-8 (95.27), C-9 (157.42), C-10 (105.91), C-1' (123.34), C-2' (112.71), C-3' (151.78), C-4' (147.23), C-5' (119.42), C-6' (123.34), C-1" (100.37), C-2" (73.56), C-3" (76.06), C-4" (70.03), C-5" (76.72), C-6" (66.50), C-1'" (100.99), C-2'" (71.20), C-4'" (72.51), C-5'" (68.80), C-6'" (18.27). On the basis of the above mentioned data, compound (5) was elucidated as chrysoeriol-7-O-rutinoside (Agrawal, 1989). It is the first report for its isolation from the plant.

Compound (6) was obtained as yellow amorphous powder (24 mg, MeOH) attained deep purple colour under UV and a yellow colour with AlCl₃ spray. UV l_max nm: MeOH (253, 357), +NaOMe (272, 400), +NaOAc (272, 380), +NaOAc/H₃BO₃ (260, 375), +AlCl₃ (275, 433), +AlCl₃/HCl (273, 404). ¹H NMR (400 MHz, DMSO-d₆) d ppm: 6.19 (1H, d, J= 2, H₆), 6.38 (1H, d, J= 2, H₈), 6.84 (1H, d, J= 8.4, H_5'), 7.54 (1H, d, J= 8.4, H_6'), 7.55 (1H, d, J= 2, H_2'), 5.34 (1H, d, J= 7.2, H_1"), 3.05-5.33 (m, sugar protons), 4.39 (1H, d, J= 0.5, H_1"'), 0.99 (3H, d, J= 6.3, H_6'"), 12.60 (1H, s, 5-OH). ¹³C NMR (100 MHz, DMSO-d₆) d ppm: C-2 (156.88), C-3 (133.66), C-4 (177.78), C-5 (161.58), C-6 (99.16), C-7 (164-52), C-8 (94.11), C-9 (157.13), C-10 (104.37), C-1' (121.58), C-2' (115.65), C-3' (145.15), C-4' (148.82), C-5' (116.64), C-6' (122.09), C-1" (101.54), C-2" (74.49), C-3" (76.24), C-4" (70.40), C-5" (75.79), C-6" (67.41), C-1'" (101.13), C-2'" (70.77), C-3'" (70.95), C-4'" (72.25), C-5'" (68.66), C-6'" (18.07). On the basis of the above mentioned evidence, compound (6) was elucidated as quercetin-3-O-aL-rhamnopyranosyl-(1-6)-b-D-glucopyranoside (rutin) (Mabry et al. and 1970, Agrawal, 1989). It is the first report for its isolation from the plant.

Table 1: Serum ALT (U/L), AST (U/L) and total bilirubin (mg/dL) values in different experimental groups.

Notes: Values are means bearing different superscripts within same raw were significantly different (P£ 0.001).

Table 2: Serum urea (mg/dL) and creatinine (mg/dL) values in different experimental groups.

Notes:        Values bearing different superscripts within same raw were significantly different (P£ 0.001).

Fig.1: Photomicrograph liver section, (A) from negative control (Tween) group showing normal histological appearance, notice the central vein and hepatic cords. (B) from reference group (Silymarin) showing mild vacuolar degeneration of the hepatocytes. (C) from CCl₄ group showing marked fatty degeneration in the hepatocytes. (D) from total methanolic extract 200mg treated group showing moderate fatty degeneration of the hepatocytes.

Fig.2: Photomicrograph liver section, (A) from total methanolic extract 400mg treated group showing minute foci of hepatocytes necrosis surrounding the centeral vein. (B) from CHCl₃ extractive 200mg treated group showing very mild vacuolar degeneration of the hepatocytes. (C) from CHCl₃ extractive 400mg showing very mild vacuolar degeneration of the hepatocytes. (D) from EtoAc extractive 200mg showing normal hepatocytes with vesicular nuclei and acidophilic cytoplasm. H&E. X 40.

Fig.3: Photomicrograph of liver section (A) from EtoAc extractive 400mg treated group showing congestion of the centeral vein with very mild vacuolar degeneration of the hepatocytes. (B) from n-butanol  extractive 200 mg treated group showing marked vacuolar degeneration of the hepatocytes and periaciner necrobiosis. (C) from n-butanol extractive 400mg treated group showing congestion of the centeral vein and marked vacuolar degeneration of the hepatocytes. H&E. X 40.

  Fig. 4:

(1)                                                     (2)

  (3)                                                     (4)

(5)

(6)

DISCUSSION

Our study revealed that treatment with ethyl acetate extractive of Cassia occidentalis L. either in a dose 200 or 400 mg/kg B.W. gave the best results in the liver function parameters in the form of marked significant decrease in ALT, AST and total bilirubin concentrations in comparison with CCl₄ treated group, and had no significant changes in values of ALT, AST and bilirubin in comparison with Silymarin treated group. Similarly, previous studies revealed significant results. Bhakta et al. (1999) studied the hepatoprotective effect of the n-heptan extractive of Cassia fistula leaves (400 mg/kg B.W.) in rats by inducing hepatoxicity with carbon tetrachloride: liquid paraffin (1:1). He found that the extract (400 mg/kg B.W) has been showed to posses protective effect by lowering the serum levels of (SGOT and SGPT), bilirubin and alkaline phosphatase (ALP). Jafri et al. (1999) also studied the effect of aqueous ethanolic extract of Cassia occientalis leaves and reported an improvement in liver function parameters (AST, ALT and alkaline phosphatase) after liver damage induced by paracetamol in rats.

The histopathological examination of the liver in different treated groups demonstrated that CCl₄ induced sever pathological changes either in the form of fatty changes or necrobiosis of the hepatocytes. These changes were subsided in Silymarin and other extractive treated groups. The treatment with ethyl acetate extractive of Cassia occidentalis aerial parts either in a dose 200 or 400 mg/kg B.W. gave the best improvement, in which the hepatic tissue appeared more or less normal. In Silymarin, methanolic extract and chloroform extractive treated groups, there were mild pathological changes in the form of congestion of vasculature and vacuolar degeneration of hepatocytes. While in the group which treated with n-butanol extractive, no histological improvement could be detected. From our results, it was clear that the histopathological findings of liver examination was positively correlated and supported the biochemical analysis of liver function parameters in different treated groups.

Our results of kidney function tests showed non significant decrease in serum urea values in all treated groups in comparison with CCl₄ treated group except the groups which treated with 200 and 400 mg/kg B.W. EtOAc extractive showed significant decrease in urea concentration in comparison with CCl₄ received group, and had no significant changes in comparison with Tween and Silymarin groups. There was significant decrease in serum creatinine levels in groupstreated with 200 and 400 mg/kg B.W. of EtOAc extractive groups in comparison with CCl₄ received group, and showed no significant changes in comparison with Tween and Silymarin groups. The other groups (Total methanolic, CHCl₃ and n-butanol extractive) showed non significant decrease in serum creatinine levels in comparison with CCl₄ received group. Mirtes et al. (2011) demonstrated that oral administration with hydroalcoholic extract of Cassia occidentalis for 30 days (0.10, 0.50 and 2.5 g/kg) in male and female rats did not changes the biochemical profile (creatinine and urea) which is good indicators of kidney functions and they reported that Cassia occidentalis extract did not induce any damage to the kidney and it is not toxic and safety for use by human. Also similar results was obtained by Hilaly et al. (2004). He found normal kidney function after oral sub-acute treatment by Cassia occidentalis extract. Fid'ele Ntchapda et al. (2015) revealed that subchronic treatment with aqueous extract of Cassia occidentalis leaves 400 mg/kg B.W. for 7 days in rats improved the kidney function index of urea and creatinine. The hepatoprotective and nephropotective effects Cassia occidentalis extracts could be attributed to its antioxidant activity (Usha et al., 2007 and Silva et. al., 2008). The histopathological findings in our results confirmed these effects. The treatment with ethyl acetate extractive of Cassia occidentalis either in a dose 200 or 400 mg/kg B.W. gave the best protection.

CONCLUSION

Cassia occidentalis ethyl acetate extractive have hepatoprotective and nephroprotective effects. This could be attributed to the antioxidant activity of its phenolic contents (compounds 1-6). Although the ethyl acetate extractive of Cassia occidentalis could be considered as a promising source of natural hepatoprotective agents, further application in medical practice should be confirmed by conveying pharmacological and placebo controlled clinical studies.

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