The potential protective effect of Physalis peruviana L. against carbon tetrachloride-induced hepatotoxicity in rats is mediated by suppression of oxidative stress and downregulation of MMP-9 expression

Abstract

The active constituent profile in Cape gooseberry (Physalis peruviana L.) juice was determined by GC-MS. Quercetin and kaempferol were active components in the juice. In this study we have evaluated its potential protective effect on hepatic injury and fibrosis induced by carbon tetrachloride (CCl4). Twenty-eight rats divided into 4 groups: Group I served as control group, and Group II received weekly i.p. injection of 2 mL CCl4/kg bwt for 12 weeks. Group III were supplemented with Physalis juice via the drinking water. The animals of Group IV received Physalis juice as Group III and also were intraperitoneally injected weekly with 2 mL CCl4/kg bwt for 12 weeks. Hepatoprotective effect was evaluated by improvement in liver enzymes serum levels, reduction in collagen areas, downregulation in expression of the fibrotic marker MMP-9, reduction in the peroxidative marker malonaldehyde and the inflammatory marker nitric oxide, and restoration of the activity of antioxidant enzymatic and nonenzymatic systems, namely, glutathione content, superoxide dismutase, catalase, glutathione-S-transferase, glutathione peroxidase, and glutathione reductase activities. The results show that the potential hepatoprotective effects of Physalis peruviana may be due to physalis acts by promotion of processes that restore hepatolobular architecture and through the inhibition of oxidative stress pathway.

Figure 1

DPPH, superoxide, thiobarbituric acid reactive substances, and nitric oxide radicals scavenging activities of physalis juice and Vit. C. IC 50 values denote the concentration of sample which is required to scavenge 50% of the respective free radicals. Data are represented as mean ± SEM of two independent experiments each performed in triplicate as published in Abdel Moneim and El-Dieb [27].

Figure 2

GC-MS chromatogram of physalis juice.

Figure 3

Photomicrographs for liver sections stained with hematoxylin and eosin (H&E). (a) Control rat showing the normal hepatocytes architecture. (b) Rats liver treated with CCl₄ showing severe hepatic lesions, degenerated and ballooned/necrotic hepatocytes with lymphocyte cells infiltration (corner). (c) Liver treated with physalis similar to control group. (d) Rat liver treated with physalis + CCl₄ showing maintained hepatic architecture, with minimal damage. Original magnification 400x.

Figure 4

Photomicrographs of liver tissue stained with Sirius Red showing. (a) Control group showing absence of collagen fibers. (b) CCl₄ group had developed extensive fibrosis in the periportal area and more fibrillar collagen deposition. (c) Physalis group showing absence of collagen fibers. (d) Physalis + CCl₄ group showing sporadic, small fibrotic lesions in the periportal zone and reduction in collagen fibers deposition. Original magnification 200x.

Figure 5

Relative quantification using RT-qPCR of mRNA expression of iNOS, GPx, and GRd genes in liver of rats treated with CCl₄ and physalis juice. Values are means ± SEM (n = 7). ^a P < 0.05, significant change with respect to Group I; ^b P < 0.05, significant change with respect to Group II.

Figure 6

Effects of physalis juice on superoxide dismutase and catalase inhibition in different brain regions of rats treated with CCl₄. Values are means ± SEM (n = 7). ^a P < 0.05, significant change with respect to Group I; ^b P < 0.05, significant change with respect to Group II.

Figure 7

The expression and specific tissue distribution of MMP-9. (a) The liver sections from control rats showed low MMP-9 immunopositive reaction in the hepatic cells. (b) In the livers of rats receiving CCl₄ for 12 weeks, high MMP-9 expression. (c) Rats receiving physalis juice for 12 weeks show weak immunopositive reaction for MMP-9 expression in hepatic cells. (d) The livers of rats receiving physalis juice concurrently with CCl₄ showed staining pattern similar or more than to the control animals. Original magnification 400x.