Abstract
Effect and Mechanism of microRNA in Exocrine Secreted by Hepatitis B Infected Hepatocytes on Activation of Hepatic Stellate Cells and Hepatic Fibrosis
Department of Hepatobiliary and Pancreatic Surgery, The Affiliated Calmette Hospital of Kunming Medical University and The First Hospital of Kunming, Kunming, Yunnan Province 650011, China
Correspondence Address:
Department of Hepatobiliary and Pancreatic Surgery, The Affiliated Calmette Hospital of Kunming Medical University and The First Hospital of Kunming, Kunming, Yunnan Province 650011, China
To explore the activation effect and mechanism of exocrine secreted by HepG2.2.15 cells on hepatic stellate cells. The exosomes from HepG2.2.15 cell culture medium and HepG2 cell culture medium were co-cultured with hepatic stellate cells respectively. The activation of hepatic stellate cells after co-culture was detected by immunofluorescence, oil red staining, scratch test, transwell, EDU, cell counting kit-8, flow apoptosis and Western blotting. The microRNA and downstream pathways which may lead to the activation of hepatic stellate cells were extracted from HepG2.2.15 cell culture medium and HepG2 cell culture medium by exosome sequencing, which were verified by quantitative polymerase chain reaction and Western blotting. After coculture with hepatic stellate cells, exosomes secreted by HepG2.2.15 cells and exosomes secreted by HepG2 cells and hepatic stellate cells, the proliferation, migration and cell activity of hepatic stellate cells were enhanced. Quantitative polymerase chain reaction detection showed that the expression of microRNA1269a in exosomes secreted by HepG2.2.15 cells co-cultured with hepatic stellate cells and exosomes secreted by HepG2 cells increased compared with hepatic stellate cells co-cultured with hepatic stellate cells, while the expression of C-mesenchymal-epithelial transition factor in stromal epidermis decreased. The hepatic stellate cells of exosomes secreted by HepG2.2.15 cells co-cultured with hepatic stellate cells and exosomes secreted by HepG2 cells and hepatic stellate cells decreased while the Yes-associated protein increased compared with that of C-mesenchymal-epithelial transition factor and phosphorylated Yes-associated protein. The exocrine secreted by HepG2.2.15 cells promote the activation of hepatic stellate cells. The mechanism may be that the exocrine carrier microRNA-1269a secreted by HepG2.2.15 is down-regulated by hepatic stellate cells, which downregulates c-mesenchymal-epithelial transition factor messenger ribonucleic acid, resulting in the decrease of C-mesenchymal-epithelial transition factor receptors on the surface of hepatic stellate cells, resulting in the decrease of Yes-associated protein phosphorylation and promoting the proliferation and activation of hepatic stellate cells.
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