Effect And Possible Mechanism Of Polidocanol On Proliferation And Apoptotic Of Lymphatic Endothelial Cells

Objective: To investigate the effects of polidocanol on the proliferation and apoptosis of lymphatic endothelial cells under the condition of 0.5%FBS,and to explore the possible regulatory mechanism of polidocanol on the remodeling of malformation lymphatic vessels and the proliferation and apoptotic of lymphatic endothelial cells.Methods: Lymphatic endothelial cells were cultured by trypsin digestion;Tunnel fluorescence test was used to measure cell apoptosis;Brd U was used to determine cell proliferation;CCK8 was used to determine cell metabolic activity;Western immunoblot test(Western-blot,WB)was used to detect the levels of apoptotic proteins Bax,Bcl 2,Caspase9 and proliferation proteins PKC and P-PKC.Results:1.FBS promotes lymphatic endothelial cells proliferation in a concentration-dependent manner and increases lymphatic endothelial cell proliferation activity;2.Polidocanol inhibits the proliferation and metabolic activity of lymphatic endothelial cells under 0.5% FBS in a concentration-dependent manner;3.0.003% polidocanol can significantly promote the apoptosis of lymphatic endothelial cells under 0.5% FBS;4.Compared with the control group,the expression levels of apoptosis proteins Bax,Bcl-2,and Caspase9 in the 0.5% FBS group were not significantly different(p> 0.05);compared with the 0.5% FBS group,the expression levels of Bax,Bcl-2 and Caspase9 in 0.5% FBS + 0.003% polidocanol group were significantly increased(p <0.01);5.0.5% FBS can induce PKC phosphorylation(P-PKC)of lymphatic endothelial cells in a time-dependent manner.Compared with the control group(0 min),P-PKC levels increased significantly at 10 min(p <0.01),and reached a peak at 30 min(p<0.01);6.0.003% polidocanol can significantly reduce the P-PKC level of lymphatic endothelial cells which induced by 0.5% FBS(p <0.01).There was no statistically significant difference in P-PKC levels between the 0.5% FBS + 0.003% polidocanol group and 0.5% FBS + Ly333531(PKC inhibitor)group(p> 0.05);there was no significant difference in PKC levels among the groups(p> 0.05);7.Compared with the control group,the proliferation and cell metabolic activity of lymphatic endothelial cells were significantly increased under the condition of 0.5%FBS(P <0.01);compared with the 0.5% FBS group,Compared with the 0.5% FBS group,the proliferation and cell metabolic activity of lymphatic endothelial cells in the 0.5% FBS + 0.003% polidocanol group were significantly reduced(p <0.01);There was no statistically significant difference in lymphatic endothelial cell proliferation and cell metabolic activity between the 0.5% FBS + 0.003% polidocanol group and 0.5% FBS + Ly333531(PKC inhibitor)group(p> 0.05).Results:1.Polidocanol inhibits the proliferation of lymphatic endothelial cells under the condition of 0.5% FBS,while reducing the cell proliferation activity;2.Polidocanol promotes apoptosis of lymphatic endothelial cells under the condition of 0.5% FBS;3.Polidocanol can induce the expression of apoptosis proteins Bax and Caspase9 in lymphatic endothelial cells under the condition of 0.5% FBS,thereby activating the endogenous apoptosis pathway and promoting apoptosis;4.Polidocanol inhibits the phosphorylation of the proliferation protein PKC;5.Polidocanol can reshape malformation lymphatic vessels by regulating lymphatic endothelial cell proliferation and apoptosis.