基于PINK1/Parkin信号通路探讨芪莲益髓清毒颗粒对急性髓系白血病Molm-13细胞线粒体自噬的调控作用

Exploration on mitophagy regulatory effect of Qilian Yisui Qingdu Granules on acute myeloid leukemia Molm-13 cell line based on PINK1/Parkin signaling pathway

目的:探讨芪连益髓清毒颗粒(QLYS)通过PINK1/Parkin信号通路对急性髓系白血病(AML)Molm-13细胞线粒体自噬的调控作用。方法:体外培养Molm-13细胞株,大鼠灌胃QLYS后制备含药血清。采用空白血清和不同浓度的QLYS含药血清分别干预Molm-13细胞,观察各组细胞的增殖情况、ATP水平、ROS水平,PINK1、Parkin、P62和LC3的变化水平。此外,采用MDC法检测自噬荧光强度,并用自噬抑制剂3-MA进行反向验证。结果:QLYS含药血清可显著抑制Molm-13细胞增殖(P<0.05,P<0.01),提高ROS水平和MDC荧光强度(P<0.01,P<0.05),降低ATP水平(P<0.05,P<0.01),增强PINK1、Parkin和LC3表达(P<0.05,P<0.01),降低P62表达(P<0.01),3-MA可降低QLYS含药血清诱导的自噬水平(P<0.05)。结论:QLYS可通过激活PINK1/Parkin信号通路促进Molm-13细胞线粒体自噬。

Objective:To investigate the regulatory effect of Qilian Yisui Qingdu Granules(QLYS)on mitophagy in the acute myeloid leukemia(AML)cell line Molm-13 based on PINK1/Parkin signaling pathway.Methods:The Molm-13 cell line was cultured in vitro,and the medicated serum was extracted from rats after gavage with QLYS.After Molm-13 cells were exposed to blank serum and different concentrations of QLYS-medicated serum,to observe cell proliferation rate,ATP levels,ROS levels,changes in the levels of PINK1,Parkin,P62,and LC3.In addition,the MDC method was used to measure the autophagy fuorescence intensity and validated the results with the autophagy inhibitor 3-MA.Results:QLYS-medicated serum inhibited Molm-13 cell proliferation(P<0.05,P<0.01),increased ROS levels and MDC fluorescence intensity(P<0.01,P<0.05),decreased ATP levels(P<0.01,P<0.05),increased PINK1,Parkin,and LC3 expressions(P<0.05,P<0.01),and decreased P62 expression(P<0.01).3-MA could reduce the autophagy level induced by QLYS-medicated serum(P<0.05).Conclusion:QLYS can promote Molm-13 cells mitophagy by activating the PINK1/Parkin signaling pathway.