LPS promotes autophagy in rat hepatic stellate cell line HSC-T6
Related Articles |
Objective To investigate the effects of lipopolysaccharide (LPS) on autophagy in rat hepatic stellate cells (HSC-T6) and involvement of NF-?B pathway in it. Methods 1) HSC-T6 cells were treated with LPS at the concentraction of 0, 0.01, 0.1, 1, and 10 mg/L for 0, 3, 6, 12, 18, and 24 h respectively, Microtubule-associated protein light chain Ⅱ (LC3Ⅱ) and Beclin1 levels were detected by Western blot; 2) HSC-T6 cells were randomized into groups of control group, LPS group, PDTC group, LPS+PDTC group, PMA group and LPS+PMA group, Western blot assay was used to measure the levels of LC3Ⅱand Beclin1, immunofluoresence was used to measure NF-?B P65 intracellular distribution. The levels of hydroxyproline (Hyp) were determined by the method of colorimetry and the levels of laminin (LN) and hyaluronic acid (HA) were determined by ELISA in the culture supernatants after corresponding processing. Results The levels of LC3Ⅱ and Beclin1 increased significantly after HSC-T6 cells were treated with LPS at the concentraction of 0.1 mg/L for 6 h and the levels of Hyp, LN, and HA in the culture supernatants increased remarkably as well (P<0.05). PDTC pretreatment increased the levels of LC3Ⅱ and Beclin1 in the LPS-treated HSC-T6 cells while decreased the levels of Hyp, LN, and HA significantly (P<0.05). PMA pretreatmen decreased the levels of LC3Ⅱ and Beclin1 in the LPS-treated HSC-T6 cells while increased the levels of Hyp, LN, and HA (P<0.05). Conclusions LPS can promote autophagy and activation of NF-?B pathway in HSC-T6 cells. Activation of NF-?B pathway may inhibit the LPS-induced autophagy of HSC-T6 cells.