小檗碱下调NLRP3减轻PC12细胞缺氧缺糖/复氧复糖损伤的作用

Berberine down-regulates NLRP3 to reduce oxygen-glucose deprivation/reoxygenation injury in PC12 cells

目的探讨小檗碱通过抑制NLRP3/Caspase-1信号通路降低神经元焦亡的作用机制。方法以大鼠PC12培养细胞构建缺氧缺糖/复氧复糖(OGD/R)诱导的细胞模型,实验分组为正常对照组、OGD/R损伤组、小檗碱低、高剂量组。MTT检测细胞活性(LDH)、TUNEL法检测细胞凋亡、免疫荧光检测GSDMD表达、酶联免疫法检测IL-1β及IL-18的分泌、Western blotting检测NLRP3、GSDMD、Caspase-1蛋白表达。结果经小檗碱处理后,与OGD/R损伤组比较:(1)神经元细胞活性明显下降(P<0.01);(2)细胞凋亡减少(P<0.01);(3)GSDMD荧光表达下降(P<0.01);(4)IL-1β及IL-18表达下降(P<0.01);(5)NLRP3、GSDMD、Caspase-1蛋白表达均有不同程度降低(P<0.01)。结论小檗碱能够抑制OGD/R诱导的神经元细胞损伤,其机制可能为通过NLRP3/Caspase-1信号通路抑制OGD/R诱导的细胞焦亡。

Objective To investigate the mechanism of action of berberine in reducing neuronal pyroptosis by inhibiting the nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)/caspase-1 signaling pathway.Methods A PC12 cell model of oxygen-glucose deprivation/reoxygenation(OGD/R)injury was prepared.The control group,OGD/R injury group,and low-and high-dose berberine groups were set up.Cell activity was measured using MTT assay.Cell apoptosis was measured using TUNEL assay.The expression of gasdermin D(GSDMD)was determined using immunofluorescence assay.The levels of interleukin-1β(IL-1β)and IL-18 were measured by enzyme-linked immunosorbent assay.The protein expression levels of NLRP3,GSDMD,and caspase-1 were measured by Western blotting.Results Compared with the OGD/R injury group,cells treated with berberine showed significantly decreased activity,apoptosis,GSDMD fluorescence intensity,and expression levels of IL-1β,IL-18,NLRP3,GSDMD,and caspase-1(all P<0.01).Conclusion Berberine can inhibit OGD/R-induced neuronal cell injury,which may be through downregulating the NLRP3/caspase-1 signaling pathway to inhibit OGD/R-induced cell pyroptosis.