Abstract

Liver Fibrosis is currently one of the top ten cause of death worldwide. Final phase of the liver fibrosis is cirrhosis, in which architecture of the liver is disrupted and extracellular matrix (ECM) is deposited. Whole-organ allograft transplantation is now a days the available treatment of last stage liver disease. Stem cells transplantation using Mesenchymal Stem Cells (MSCs) is an alternative therapy in place of organ transplant but due to incapacity of stem cells to endure the oxidative stress and inflammatory responses in the damage site affecting the healing process of this therapy. Present study was aimed to enhance the therapeutic potential of MSCs by combined therapy along with novel synthetic compounds of benzimidazol derivatives. Eighteen compound series (benzimidazole derivatives) were screened against liver fibrosis using In Vitro CCl₄ induced injury model of hepatocytes. IC50 were calculated on the basis of LDH assay and cell viability assay. Among eighteen, compounds 10, 14 and 18 were selected on the basis of IC50 value in which compound 10 was most potent with lowest IC50 value in LDH assay (8.399 0.23um) and cell viability assay (4.73 0.37um). In the next steps these compounds were used combined with MSCs using In Vitro hepatocytes injury model and In Vivo rat fibrotic model. Effect of MSCs+Compounds treatment on injured hepatocytes were evaluated using LDH assay, cell viability assay, GSH assay and real time PCR  analysis and Immunostaining for Caspase-3. Treatment effect of MSCs+Compounds on liver fibrosis was measured by histopathology, PAS staining, TUNEL Assay, Masson Trichrome Staining, real time PCR and Biochemical assay. Significant reduction in LDH level, Caspase-3 and apoptotic marker genes were noted in MSCs+Compounds treated injured hepatocytes. Cell viability and antiapoptotic marker were upregulated in MSCs+Compound treated hepatocytes. In Vivo data also showed increased homing of MSCs along with compounds after transplantation into liver fibrotic model as confirmed the results of glycogen restoration and reduction in collagen, ALP, AST, ALT and bilirubin level. Real time PCR analysis and TUNEL assay results are also support our study. Therefore, it is concluded that Compounds 10, 14 and 18 can be used in combination with MSCs for the reduction of liver fibrosis.

Key words; Stem cells, Benzimidazole, CCl4, hepatocytes, rats