补肾活血颗粒含药血清对MPP~+诱导的PC12细胞Nrf2/NLRP3/Caspase-1通路的影响  

Effects of drug-containing serum of Bushen Huoxue Granules on the Nrf2/NLRP3/Caspase-1 pathway in MPP+-induced PC12 cells

作　　者：陈琦[1,2] 王鹏 谌盈帆 李绍旦 杨明会[1] CHEN Qi;WANG Peng;CHEN Yingfan;LI Shaodan;YANG Minghui(Senior Department of Traditional Chinese Medicine,the Sixth Medical Center of PLA General Hospital,Beijing 100048,China)

机构地区：[1]解放军总医院第六医学中心中医医学部,北京100048 [2]解放军医学院

出　　处：《环球中医药》2025年第4期677-684,共8页Global Traditional Chinese Medicine

基　　金：国家自然科学基金(82205231)。

摘　　要：目的基于Nrf2/NLRP3/Caspase-1通路探讨补肾活血颗粒(Bushen Huoxue Granule,BSHX)含药血清对MPP^(+)诱导的PC12细胞的影响。方法MPP^(+)诱导PC12细胞建立帕金森病细胞模型,制备空白血清和BSHX含药血清,设立空白对照组、MPP^(+)组、BSHX组、siNrf2组、BSHX+SiNrf2、siCaspase-1和BSHX+siCaspase-1组。采用CCK-8法检测细胞活力,免疫印迹实验、聚合酶链式反应法分别检测PC12细胞核因子E2相关因子2(nuclear factor erythroid 2 related factor,Nrf2)、核苷酸结构域样受体蛋白3[nucleotide-binding domain(NOD)-like receptor protein 3,NLRP3]、天冬氨酸蛋白水解酶1(Cysteinyl aspartate specific proteinase 1,Caspase-1)蛋白和mRNA表达;免疫荧光法检测α-突触核蛋白(α-synuclein,α-syn)表达;酶联免疫吸附试验检测白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-18(interleukin-18,IL-18)表达。结果与空白对照组相比,MPP^(+)组、siNrf2组PC12细胞Nrf2蛋白和mRNA表达降低,NLRP3、Caspase-1蛋白和mRNA表达升高,α-syn、IL-1β、IL-18表达升高(P<0.01)。与MPP^(+)组相比,BSHX组、BSHX+SiNrf2组、BSHX+siCaspase-1组PC12细胞Nrf2蛋白和mRNA表达升高,NLRP3、Caspase-1蛋白和mRNA表达降低,α-syn、IL-1β、IL-18表达降低(P<0.01)。结论补肾活血颗粒含药血清可以通过激活Nrf2抑制NLRP3和Caspase-1表达,减少IL-1β、IL-18分泌,从而减轻PC12细胞炎症损伤和减少α-syn聚集。Objective To investigate the effects of Bushen Huoxue Granules(BSHX)-containing serum on MPP+induced PC12 cells based on the Nrf2/NLRP3/Caspase1 pathway.Methods A Parkinson’s disease cell model was established by inducing PC12 cells with MPP+.Blank serum and BSHX-containing serum were prepared.Experimental groups included the control group,the MPP+group,the BSHX group,the siNrf2 group,the BSHX+siNrf2 group,the siCaspase-1 group,and the BSHX+siCaspase-1 group.Cell viability was assessed using the CCK-8 assay.The protein and mRNA expression levels of nuclear factor erythroid 2 related factor(Nrf2),nucleotide-binding domain(NOD)-like receptor protein 3(NLRP3),and Cysteinyl aspartate specific proteinase 1(Caspase-1)in PC12 cells were measured by the methods of Western blot and polymerase chain reaction.Immunofluorescence was used to detectα-syn expression,and enzyme-linked immunosorbent assay was used to assess IL-1βand IL-18 levels.Results Compared with the control group,the MPP+and siNrf2 groups exhibited decreased Nrf2 protein and mRNA expression in PC12 cells,while NLRP3 and Caspase-1 protein and mRNA expression increased,andα-synuclein(α-syn),interleukin(IL)-1β,and IL-18 levels were increased(P<0.01).Compared with the MPP+group,the BSHX group,the BSHX+SiNrf2 group and the BSHX+siCaspase-1 group showed increased cell survival rate,elevated Nrf2 protein and mRNA expression,reduced NLRP3 and Caspase-1 protein and mRNA expression,and decreased levels ofα-syn,IL-1β,and IL-18(P<0.01).Conclusion BSHX-containing serum alleviates inflammation andα-syn aggregation in PC12 cells by activating Nrf2,inhibiting NLRP3 and Caspase-1 expression,and reducing IL-1βand IL-18 secretion.

关 键 词：补肾活血颗粒 含药血清 核因子E2相关因子2 核苷酸结构域样受体蛋白3 天冬氨酸蛋白水解酶1 

分 类 号：R285.5[医药卫生—中药学]