Hepatoprotective and Anti-Inflammatory Activities of the Cnidoscolus chayamansa (Mc Vaugh) Leaf Extract in Chronic Models

Abstract

Previous report described that CHCl₃:MeOH extract of C. chayamansa leaves and pure compounds (moretenol, moretenyl acetate, kaempferol-3,7-dimethyl ether, and 5-hydroxy-7-3',4'-trimethoxyflavanone) showed important topical and systemic anti-inflammatory activity in acute model, as well as in vitro antimycobacterial and antiprotozoal activities. In this paper, we describe the in vivo hepatoprotective and anti-inflammatory effects of the CHCl₃:MeOH extract in chronic model and the isolation of additional compounds (moretenone and lupeol acetate). The hepatoprotective activity was determined at 39 days using Balb/c mice with liver damage induced with an antitubercular drug (RIF/INH/PZA). The anti-inflammatory activity was evaluated in a chronic model induced with CFA and in two acute models (TPA and carrageenan). In addition, moretenone and lupeol acetate were isolated and identified by spectroscopic data. Lupeol acetate is a main compound present in fractions 14-42, and this fraction was the majority fraction from the extract; from this moretenone was obtained. In animals with liver damage, the extract at 200 and 400 mg/kg induced better body weight gain with respect to positive control (Silymarin); in addition, the mice that received the extract at 200 mg/kg and Silymarin exhibited slight steatosis; however, the animals' livers at 400 mg/kg did not show steatosis. The extract and fractions 14-42 showed a good anti-inflammatory activity by TPA model (DE₅₀ = 1.58 and 1.48 mg/ear) and by carrageenan model (DE₅₀ = 351.53 and 50.11 mg/kg). In the chronic inflammatory test, the extract at three doses revealed a similar effect to that of phenylbutazone, although the body weight gain was better in animals that received the extract at 900 mg/kg. Conclusion. The CHCl₃:MeOH extract showed significant anti-inflammatory and good hepatoprotective effect on chronic models. The fraction containing moretenone and lupeol acetate as main compounds was more active than extract in both acute models of inflammation.

Figure 1

X-ray crystal structures of moretenone.

Figure 2

Effect of C. chayamansa extract on edema development (mm) during experimental chronic model in mice. Data shown as mean ± standard error (SEM). Treatments were administered daily by i.g. route from day 7 to 28. Two-way analysis of variance (ANOVA) of repeated measures (RM); post hoc Student-Newman-Keuls (SNK) (p≤0.05). ^avs vehicles; ^bvs CFA control; ^cvs CFA+PBZ; ^dvs CFA+ CnCM 200 mg/kg; ^evs CFA+ CnCM 450 mg/kg; ^●vs day 1; ∗vs day 7; ^▲vs day 14; ^■vs day 21. CFA: Complete Freund's adjuvant; PBZ: phenylbutazone; n= 7.

Figure 3

Effect of C. chayamansa extract on BW gain (g) during experimental chronic anti-inflammatory model in mice. Data shown as mean (±) standard error (SEM). Treatments were administered daily by intragastric route from day 7 to day 28. Two-way analysis of variance (ANOVA) of repeated measures (RM); post hoc Student-Newman-Keuls (SNK), (p≤0.05). ^avs vehicles; ^bvs CFA control; ^cvs CFA+PBZ; ^dvs CFA+ CnCM 200 mg/kg; ^evs CFA+ CnCM 450 mg/kg; ^●vs day 1; ∗vs day 7; ^▲vs day 14; ^■vs day 21. CFA: Complete Freund's adjuvant; PBZ: phenylbutazone; CnCM: CHCl₃:MeOH extract, n= 7.

Figure 4

Effect of C. chayamansa extract on BW gain (g) during hepatotoxicity induced by antiTB drugs in Balb/C mice. Data presented as mean (±) standard error (SEM). Statistical analysis of variance (ANOVA); bifactorial of repeated measures (RM); post hoc Student-Newman-Keuls (SNK) test (p<0.05); ^avs AntiTB; ^bvs AntiTB + Silymarin 2.5 mg/kg; ^cvs AntiTB + CnCM 200 mg/kg; ^dvs AntiTB + CnCM 400 mg/kg; AntiTB, antitubercular drugs (RIF/INH/PZA); n=7.

Figure 5

Effect of C. chayamansa extract over relative liver weight of Balb/C mice with hepatotoxicity induced by antiTB drugs. Data presented as mean (±) standard error (SEM). Statistical Analysis Kruskal-Wallis (ANOVA on Ranks) and post hoc Student-Newman-Keuls (SNK) test (p<0.05); ^avs AntiTB; ^bvs AntiTB + Silymarin 2.5 mg/kg; ^cvs AntiTB + CnCM 200 mg/kg; ^dvs AntiTB + CnCM 400 mg/kg; AntiTB, antitubercular drugs (RIF, INH, PZA) mixture; CnCM: CHCl₃:MeOH extract, n=7.

Figure 6

Histological liver section from Balb/C mice with hepatotoxicity induced by antiTB drugs and treated for 39 days with the CnCM extract. Histologic liver tissue (x100) collected from mice euthanized on day 39; after the treatment period, tissue were stained with hematoxylin and eosin 16X. AntiTB group showed steatosis, Silymarin (2.5 mg/kg) and CnCM (200 mg/kg) showed a moderate steatosis, and CnCM (400 mg/kg) group showed a scare steatosis.