Effect of autophagy inhibitor 3-methyladenine on phenotype transformation and proliferation of pulmonary arterial smooth muscle cells / 实用医学杂志

Objective To evaluate the effect of autophagy inhibitor 3-methyladenine (3-MA) on phenotype transformation and proliferation of rat pulmonary arterial smooth muscle cells (PASMCs).Methods Cultured PASMCs were treated with different concentrations of 3-MA (low-dose group,2.0 mmol/L;middle-dose group,4.0 mmol/L;high-dose group,8.0 mmol/L;control group,0 mmol/L).The protein expression of LC3 Ⅱ,OPN and Vimentin was detected by Western blotting.Cell proliferation was detected by MTT assay.Results The autophagy of PASMCs was decreased with the increase of the concentration of 3-MA.Compared with the control group,significantly down-regulated protein expression of LC3 Ⅱ,OPN and Vimentin was observed in 3-MA-treated cells,with significantly lower proliferation activity.Conclusion The autophagy inhibitor 3-MA significantly down-regulated the expression of LC3 Ⅱ,OPN and Vimentin in PASMCs,with inhibiting the proliferation of PASMCs.

Effect of serum of patients with obstructive jaundice on myogenic differentiation of human pulmona-ry microvascular endothelial cells / 中华麻醉学杂志

Objective To evaluate the effects of the serum of patients with obstructive jaundice on myogenic differentiation of human pulmonary microvascular endothelial cells (PMVECs). Methods Hu?man PMVECs were isolated and then subcultured. The cultured PMVECs were incubated with the serum of patients with obstructive jaundice or with the serum of healthy volunteers. At 24, 48 and 72 h of incubation (T1?3), the inverted microscope was used to observe the morphology of primary PMVECs. The expression of muscular proteins ( alpha?smooth muscle actin [α?SMA ] , smooth muscle?mysion heavy chain [ SM?MHC] , capolnin) in PMVECs was detected using Western blot analysis. Results The expression of cal?ponin andα?SMA was negative, and a few SM?MHC proteins were expressed when PMVECs were incubated with the serum of healthy volunteers; the expression of calponin, α?SMA and SM?MHC was positive when PMVECs were incubated with the serum of patients with obstructive jaundice. Compared with the serum of healthy volunteers, the expression of SM?MHC was significantly up?regulated when PMVECs were incubated with the serum of patients with obstructive jaundice (P<0.05). The expression of calponin, α?SMA and SM?MHC was significantly up?regulated at T2,3 compared with that at T1 , and at T3 compared with that at T2 when PMVECs were incubated with the serum of patients with obstructive jaundice ( P<0.05) . Conclusion The serum of patients with obstructive jaundice promotes myogenic differentiation of human PMVECs, which is probably one of the mechanisms underlying intrapulmonary microvascular dilatation.