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tenantId=1146029695717560320, journalId=1315121837170065538, language=CN, title=筋骨痛消丸通过AMPK/自噬/NLRP3信号轴调控膝骨关节炎软骨退变的机制研究, columnId=null, journalTitle=中医学报, columnName=实验研究, runingTitle=null, highlight=null, articleAbstract=目的:探讨筋骨痛消丸含药血清通过腺苷一磷酸活化蛋白激酶（adenosine monophosphate activated protein kinase,AMPK）/自噬/核苷酸结合寡聚化结构域样受体蛋白3（NOD-like receptor thermal protein domain associated protein 3,NLRP3）信号轴调控膝骨关节炎（knee osteoarthritis,KOA）软骨细胞退变的作用机制。方法:取培养至第2代的人软骨细胞分为5组:空白组、模型组、筋骨痛消丸组、AMPK抑制剂组、自噬抑制剂组。空白组加入10%空白血清，模型组加入10%空白血清和10μg·L⁻¹白细胞介素-1β（interleukin-1β,IL-1β），筋骨痛消丸组加入10%筋骨痛消丸含药血清和10μg·L⁻¹IL-1β,AMPK抑制剂组加入10%筋骨痛消丸含药血清、10μg·L⁻¹IL-1β及10μmol·L⁻¹AMPK抑制剂（Dorsomorphin），自噬抑制剂组加入10%筋骨痛消丸含药血清、10μg·L⁻¹IL-1β及5 mmol·L⁻¹自噬抑制剂（3-MA），继续培养24 h。采用CCK-8检测各组细胞增殖率，通过流式细胞术检测细胞凋亡率及自噬活性，采用实时定量PCR检测AMPK、哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin,m TOR）、微管相关蛋白1轻链3B（microtubule-associated protein 1 light chain 3B,LC3B）、Beclin-1、UNC-51样自噬激活激酶1（UNC-51 like autophagy activating kinase 1,ULK1）、NLRP3、凋亡相关斑点样蛋白（apoptosis-associated specklike protein containing a CARD,ASC）、胱天冬氨酸蛋白酶-1（cysteine-requiring aspartate protease-1,Caspase-1）、IL-1β、白细胞介素-18（interleukin-18,IL-18）的mRNA表达水平，通过Western blot检测磷酸化AMPK（phospho-AMPK,p-AMPK）、Beclin-1、ULK1、LC3B-Ⅰ、LC3B-Ⅱ、磷酸化mTOR（phospho-mTOR,p-mTOR）、NLRP3、Caspase-1、IL-1β的蛋白表达水平。结果:与空白组对比，模型组细胞增殖率、细胞自噬率显著降低（P <0.05），细胞凋亡率显著升高（P <0.05）,AMPK、LC3B、Beclin-1、ULK1 mRNA和蛋白表达均显著下调（均P <0.05）,mTOR、NLRP3炎症小体相关的mRNA和蛋白表达水平均显著升高（P <0.05）;与模型组对比，筋骨痛消丸组细胞增殖率、细胞自噬率显著升高（P <0.05），细胞凋亡率显著降低（P <0.05）,AMPK、LC3B、Beclin-1及ULK1 mRNA和蛋白表达均显著升高（均P <0.05）,mTOR、NLRP3炎症小体相关的mRNA和蛋白表达水平均显著降低（P <0.05）;与筋骨痛消丸组对比，AMPK抑制剂组和自噬抑制剂组的细胞增殖率、细胞自噬率显著降低（P <0.05），细胞凋亡率显著升高（P <0.05）,AMPK抑制剂组和自噬抑制剂组AMPK、LC3B、Beclin-1、ULK1 mRNA表达均显著下调（均P <0.05）,mTOR、NLRP3炎症小体相关的mRNA和蛋白表达水平均显著上调（P <0.05）。结论:筋骨痛消丸可能通过激活AMPK/自噬轴抑制NLRP3炎性小体活化，促进KOA软骨细胞增殖和自噬，抑制凋亡，调控KOA软骨细胞退变。, authors=null, authorsList=null, authorCompany=null, correspAuthors=null, authorNote=null, correspAuthorsNote=null, 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