cGAS-STING signaling pathway mediates the effects of the abnormal fluid shear stress on apoptosis,inflammation and autophagy in nucleus pulposus cells
Objective To investigate the effects of the abnormal fluid shear stress(FSS)on apoptosis,inflam-mation and autophagy in nucleus pulposus cells(NPC)and its possible mechanisms.Methods NPC were load-ed with 24 dyn/cm2 FSS for 0,60,90,120 and 150 min respectively.Cytoskeleton staining was used to study the effect of FSS on cytoskeleton and cell morphology.5-acetylene-2'-deoxyuridine staining was used to veri-fy the proliferation ability of NPC;JC-1 mitochondrial membrane potential kit used to study the mitochondrial damage induced by FSS;Hoechst 33342 staining used to study the relationship between apoptosis and FSS,and monodansylcadaverine staining used to study the relationship between autophagy and FSS.The chang-es of pH in cell culture medium were explored to verify the relationship between FSS and inflammation.Western blotting was used to study the expression levels of cGAS-STING related proteins(cGAS,STING,TBK1),inflammation-related proteins(TNF-α,COX-2),apoptosis-related proteins(Bcl-2,Bax)and autopha-gy proteins(IL3-Ⅰ/Ⅱ,p62).Results As an intercellular mechanical stimulus,the 24 dyn/cm2 FSS could cause the morphology of NPC to change from a spindle shape to polygon shape,and cause the reorganization of cytoskeleton,decrease in cell proliferation ability,increase in the acidity of the medium,decrease of mitochondrial JC-1 multimers and increase of mitochondrial monomers.At the same time,the expressions of TNF-α,COX-2 and Bax proteins in the cells were up-regulated,the LC3Ⅱ/LC3Ⅰratio increased,the expres-sions of p62 and Bcl-2 decreased,and apoptosis,inflammation,autophagy and mitochondrial damage were observed.In addition,24 dyn/cm2 FSS could activate the cGAS-STING signaling pathway in NPC in a time-dependent manner,with the highest intensity at 120 min and treatment with the cGAS inhibitor RU.521 could alleviate the apoptosis,inflammation,autophagy and mitochondrial damage of NPC caused by 24 dyn/cm2 FSS.Conclusion FSS-induced apoptosis,inflammation and autophagy of NPC were mediated by the activa-tion of cGAS-STING signaling pathway.Inhibition of cGAS-STING signaling pathway could alleviate the cellular damage caused by the abnormal FSS.
国家科技期刊平台
NSTL
万方数据
