Study on the mechanism of Qingdu Tang in protecting the intestinal barrier and treating endotoxemia in acute-on-chronic liver failure
Objective To study the protective effect of Qingdu Tang on the intestinal barrier,and to explore the mechanism of protecting the intestinal barrier by Qingdu Tang in the treatment of endotoxemia in slow plus acute liver failure.Methods HIEC-6 cells were treated with different concentrations of lipopolysaccharides(LPS),and the effect of LPS on the survival rate of HIEC-6 cells was detected by cytotoxicity assay(cell counting kit-8,CCK-8).The mRNA expression of tight junction proteins ZO-1 and Occludin was detected by quantitative real-time fluorescence PCR(qRT-PCR),and the model of intestinal barrier damage was constructed in vitro.The effect of Qingdu Tang on the expression levels of intestinal barrier tight junction proteins ZO-1 and Occludin were detected by protein immunoblotting(western blot,WB).The mRNA expressions of pathway proteins EGFR,Zonulin and miRNA122a were detected by qRT-PCR.Results The results of CCK-8 method and qRT-PCR showed that 120 μg/mL LPS was a more appropriate concentration for constructing the intestinal barrier injury model.The results of qRT-PCR showed that 5％Qingdu Tang containing serum significantly improved the down-regulation of LPS on the mRNAs of ZO-1 and Occludin.The results of WB showed that the serum containing 5％Qingdu Tang significantly improved the down-regulation of LPS on ZO-1 and Occludin,which are tight junction proteins between intestinal epithelial cells.The result of qRT-PCR showed that Qingdu Tang significantly reduced the expression of the pathway proteins,such as EGFR,Zonulin,and miRNA-122a.Conclusion Qingdu Tang may target EGFR through miRNA-122a to regulate Zonulin protein thus regulating the expression of intestinal tight junction,protecting the intestinal barrier,and treating endotoxemia in slow plus acute liver failure.
万方数据
维普
