瑞舒伐他汀调节Nrf2/HO-1信号通路对病毒性心肌炎细胞凋亡和炎症反应影响的研究OA
Research of impacts of rosuvastatin on apoptosis and inflammatory response of viral myocarditis cells by regulating the Nrf2/HO-1 signaling pathway
目的 探讨瑞舒伐他汀调节核因子红细胞2相关因子2/血红素加氧酶-1(Nrf2/HO-1)信号通路对病毒性心肌炎细胞凋亡和炎症反应的影响.方法 心肌H9c2细胞分为空白组(常规DMEM培养基培养)、模型组[含柯萨奇病毒B3(CVB3)病毒悬液DMEM培养基培养]、实验组(含CVB3病毒悬液+10 μmol·L-1瑞舒伐他汀的DMEM培养基培养)、ML385组(含CVB3病毒悬液+5 μmol·L-1 ML385的DMEM培养基)、瑞舒伐他汀(RSV)+ML385组(含CVB3病毒悬液+10 μmol·L-1瑞舒伐他汀+5 μmol·L-1 ML385 DMEM培养基),以上各组均处理48 h后,用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法检测细胞活力、用酶联免疫吸附测定(ELISA)法检测白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)水平,用流式细胞术检测细胞凋亡,用实时荧光定量聚合酶链式反应(qRT-PCR)及蛋白质免疫印迹法分别检测Nrf2、HO-1、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)mRNA及蛋白相对表达水平表达.结果 空白组、模型组、实验组、ML385组和RSV+ML385组的细胞活力分别为(90.52±8.65)％、(36.25±3.12)％、(53.84±5.96)％、(25.17±2.48)％和(38.49±4.03)％,IL-1β 水平分别为(5.36±0.74)、(18.62±1.91)、(7.85±0.87)、(27.28±2.49)和(19.43±2.15)ng·mL-1,TNF-α 水平分别为(18.09±1.32)、(42.46±4.75)、(25.12±2.49)、(63.75±6.16)和(38.91±4.08)ng·mL-1,IL-6 水平分别为(45.82±4.19)、(121.29±13.36)、(52.48±5.64)、(176.94±18.83)和(119.06±13.57)ng·mL-1,细胞凋亡率分别为(7.15±0.62)％、(23.47±2.59)％、(15.83±1.48)％、(34.29±3.94)％和(22.64±2.73)％,Nrf2 mRNA 相对表达水平分别为1.05±0.14、0.58±0.07、0.76±0.08、0.27±0.03 和 0.53±0.07,HO-1 mRNA相对表达水平分别为 0.94±0.11、0.62±0.07、0.89±0.10、0.38±0.04 和0.60±0.07,Bcl-2 mRNA 相对表达水平分别为 0.98±0.10、0.45±0.04、0.78±0.08、0.22±0.03 和 0.48±0.06,Bax mRNA 相对表达水平分别为1.03±0.11、1.78±0.18、1.15±0.12、2.35±0.26 和 1.72±0.17;Nrf2 蛋白相对表达水平分别为 0.75±0.08、0.41±0.05、0.69±0.07、0.28±0.03 和0.47±0.05,HO-1 蛋白相对表达水平分别为 1.42±0.15、0.84±0.09、1.35±0.14、0.58±0.06和0.79±0.08,Bcl-2蛋白相对表达水平分别为1.15±0.12、0.64±0.07、0.97±0.10、0.43±0.05 和 0.68±0.07,Bax 蛋白相对表达水平分别为 0.45±0.05、0.78±0.08、0.52±0.06、1.37±0.14 和0.72±0.08.模型组的上述指标与空白组比较,实验组和ML385组的上述指标与模型组比较,RSV+ML385组的上述指标与实验组比较,在统计学上差异均有统计学意义(均P＜0.05).结论 瑞舒伐他汀可能通过激活Nrf2/H O-1信号通路,部分减轻病毒性心肌炎细胞凋亡并抑制炎症反应.
Objective To investigate the impacts of rosuvastatin on apoptosis and inflammatory response of viral myocarditis cells by regulating the nuclear factor erythroid 2-related factor 2/heme oxygenase-1(Nrf2/HO-1)signaling pathway.Methods Myocardial H9c2 cells were divided into blank group(cultured in regular DMEM medium),model group(cultured in DMEM medium containing coxsackievirus B3(CVB3)suspension),experimental group(cultured in DMEM medium containing CVB3 suspension+10 μmol·L-1 rosuvastatin),ML385 group(cultured in DMEM medium containing CVB3 suspension+5 μmol·L-1 ML385)and rosuvastatin(RSV)+ML385 group(cultured in DMEM medium containing CVB3 suspension+10 μmol·L-1 rosuvastatin+5 μmol·L-1 ML385).After 48 h of treatment in all groups,cell viability,levels of interleukin-1 β(IL-1 β),tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6),as well as apoptosis were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay,enzyme-linked immunosorbent assay(ELISA),and flow cytometry,respectively.The mRNA and protein expression levels of Nrf2,HO-1,B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax)were detected by quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot.Results The cell viability in the blank,model,experimental,ML385 and RSV+ML385 groups were(90.52±8.65)％,(36.25±3.12)％,(53.84±5.96)％,(25.17±2.48)％and(38.49±4.03)％,respectively;the IL-1 β levels were(5.36±0.74),(18.62±1.91),(7.85±0.87),(27.28±2.49)and(19.43±2.15)ng·mL-1,respectively;TNF-α levels were(18.09±1.32),(42.46±4.75),(25.12±2.49),(63.75±6.16)and(38.91±4.08)ng·mL-1,respectively;IL-6 levels were(45.82±4.19),(121.29±13.36),(52.48±5.64),(176.94±18.83)and(119.06±13.57)ng·mL-1,respectively;apoptosis rates were(7.15±0.62)％,(23.47±2.59)％,(15.83±1.48)％,(34.29±3.94)％and(22.64±2.73)％,respectively;the relative expression levels of Nrf2 mRNA were 1.05±0.14,0.58±0.07,0.76±0.08,0.27±0.03 and 0.53±0.07,respectively;the relative expression levels of HO-1 mRNA were 0.94±0.11,0.62±0.07,0.89±0.10,0.38±0.04 and 0.60±0.07,respectively;the relative expression levels of Bcl-2 mRNA were 0.98±0.10,0.45±0.04,0.78±0.08,0.22±0.03 and 0.48±0.06,respectively;the relative expression levels of Bax mRNA were 1.03±0.11,1.78±0.18,1.15±0.12,2.35±0.26 and 1.72±0.17,respectively.The Nrf2 protein expression levels were 0.75±0.08,0.41±0.05,0.69±0.07,0.28±0.03,and 0.47±0.05,respectively;the relative expression levels of HO-1 protein levels were 1.42±0.15,0.84±0.09,1.35±0.14,0.58±0.06,and 0.79±0.08,respectively;the relative expression levels of Bcl-2 protein levels were 1.15±0.12,0.64±0.07,0.97±0.10,0.43±0.05,and 0.68±0.07,respectively;the relative expression levels of Bax protein levels were 0.45±0.05,0.78±0.08,0.52±0.06,1.37±0.14,and 0.72±0.08,respectively.The differences between the model group and the blank group,between the experimental group or ML385 group and the model group,and between the RSV+ML385 group and the experimental group were all statistically significant(P＜0.05).Conclusion Rosuvastatin may partially alleviate apoptosis and inhibit the inflammatory response in viral myocarditis by activating the Nrf2/HO-1 signaling pathway.
李林;程茵;李梓楠;侯乐萍
杭州市第一人民医院城北院区药学部,浙江 杭州 310000杭州市第一人民医院城北院区药学部,浙江 杭州 310000杭州市第一人民医院城北院区药学部,浙江 杭州 310000杭州市第一人民医院城北院区药学部,浙江 杭州 310000
医药卫生
瑞舒伐他汀核因子红细胞2相关因子2/血红素加氧酶-1信号通路病毒性心肌炎凋亡炎症反应
rosuvastatinnuclear factor erythroid 2-related factor 2/heme oxygenase-1 signaling pathwayviral myocarditisapoptosisinflammatory response
《中国临床药理学杂志》 2026 (15)
2161-2168,8
浙江省医药卫生科技计划基金资助项目(2022KY279)杭州市生物医药和健康产业发展扶持专项基金资助项目(2023WJC015)
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