Effect of novel turtle shell decoction on cycle, apoptosis, CTGF/Itgα 5 pathway and expression of Col1a1 in rat hepatic stellate cells

Abstract

Objective To investigate the effects of novel turtle shell decoction (NTSD) on the cycle, apoptosis, connective tissue growth factor/integrin alpha 5 (CTGF/Itgα5) pathway and the expression of Collagen type I (Col1a1) in rat hepatic stellate cell line T6 (HSCT6). Methods HSCT6 cells was treated with NTSD containing crude drug 0.71 g/mL as experimental group, treated with turtle shell pills (TSP) at the same dose as positive control, and cultured routinely as normal control. Cell cycle and apoptosis of HSCT6 cells were tested by flow cytometry (FCM). Real time-qPCR, Western blotting and immunofluorescence assay were employed to detect the expression changes of CTGF, Itgα5 and Col1a1 genes and their coding proteins in each group of HSCT6, respectively. Results FCM showed that NTSD treatment blocked the cells at S stage, and induced cell apoptosis. Real time-qPCR, Western blotting and immunofluorescence assay revealed that NTSD can significantly down-regulate the expression of CTGF, Itgα5 and Col1a1 in mRNA and protein levels in HSC-T6 cells (P < 0.01). Conclusion NTSD treatment results in cell arrest at S phage through down-regulating CTGF/Itgα5, induction of cell apoptosis, decrease of Col1a1 expression, and then inhibition of cell growth in HSCT6 cells.

Keywords

• novel turtle shell decoction
• hepatic stellate cells
• connective tissue growth factor
• integrin alpha 5
• collagen type i
• cell cycle
• apoptosis