参元益气活血胶囊对肿瘤坏死因子α诱导小鼠心肌细胞损伤的影响及机制  

Effect and mechanism of Shenyuan Yiqi Huoxui Capsules on TNF-α-induced myocardial cell injury in mice

作　　者：娄妍 刘红旭[1] 尚菊菊[1] 周琦[1] 周明学 李享[1] 邢文龙[1] LOU Yan;LIU Hongxu;SHANG Juju;ZHOU Qi;ZHOU Mingxue;LI Xiang;XING Wenlong(Department of Cardiology,Beijing Hospital of Traditional Chinese Medicine,Capital Medical University,Beijing 100010,China;Capital Medical University,Beijing 100069;Brain and Heart Tongzhi Laboratory,Beijing Institute of Traditional Chinese Medicine,Beijing 100010)

机构地区：[1]首都医科大学附属北京中医医院心血管科,北京100010 [2]首都医科大学,北京100069 [3]北京市中医药研究所脑心同治实验室,北京100010

出　　处：《北京中医药》2024年第11期1275-1280,共6页Beijing Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目面上课题(8217151495);国家自然科学基金资助项目青年课题(82205098)。

摘　　要：目的探讨参元益气活血胶囊(SYYQ)对肿瘤坏死因子α(TNF-α)诱导的小鼠心肌细胞损伤的影响及机制。方法常规培养小鼠心肌细胞(HL-1细胞),并制备SYYQ含药血清。用0、10%、20%、50%的SYYQ含药血清干预HL-1细胞,用细胞计数试剂法(CCK-8)检测细胞存活率,根据细胞存活率筛选最佳实验浓度。将HL-1细胞随机分为3组,对照组加入DMEM培养,模型组用50 ng/mL的TNF-α刺激12 h诱导心肌损伤模型,SYYQ组在模型组基础上加入最佳实验浓度的含药血清,同步培养12 h。流式细胞术检测细胞凋亡,Real-time PCR检测细胞内核因子κB(NF-κB)、核苷酸结合寡聚域样受体蛋白3(NLRP3)、白细胞介素18(IL-18)mRNA表达,Western blotting法检测细胞内NF-κB、NLRP3、IL-18蛋白表达。结果不同浓度SYYQ含药血清干预的心肌细胞存活率分别为(53.32±5.70)%、(68.24±6.41)%、(92.62±8.56)%、(86.93±7.12)%,故将20%含药血清组作为药物干预细胞的最佳浓度。对照组、模型组、SYYQ组心肌细胞凋亡率分别为(1.57±0.18)%、(18.99±1.32)%、(6.76±0.38)%。与对照组比较,模型组心肌细胞凋亡率高(P<0.01);与模型组比较,SYYQ组心肌细胞凋亡率低(P<0.01)。与对照组比较,模型组细胞内NF-κB、NLRP3、IL-18 mRNA相对表达量高(P<0.05,P<0.01);与模型组比较,SYYQ组细胞内NF-κB、NLRP3、IL-18 mRNA相对表达量低(P<0.01)。与对照组比较,模型组细胞内NLRP3、IL-18蛋白相对表达量高(P<0.01);与模型组比较,SYYQ组细胞内NF-κB、NLRP3、IL-18蛋白相对表达量低(P<0.05,P<0.01)。结论SYYQ可能通过调控NF-κB/NLRP3/IL-18信号通路减轻TNF-α所致心肌细胞损伤。Objective To investigate the effect of Shenyuan Yiqi Huoxue Capsules(SYYQ)on tumor necrosis factorα(TNF-α)-induced myocardial cell injury in mice and its mechanism.Methods Mouse cardiomyocytes(HL-1 cells)were cultured routinely,and SYYQ-containing serum was prepared.HL-1 cells were treated with 0%,10%,20%,and 50%SYYQcontaining serum.Cell viability was measured using the cell counting kit-8(CCK-8)assay,and the optimal experimental concentration was selected based on the cell survival rate.HL-1 cells were randomly divided into three groups:the control group,which was cultured with DMEM;the model group,which was stimulated with 50 ng/mL TNF-αfor 12 h to induce myocardial injury;the SYYQ group,which was treated with the optimal concentration of drug-containing serum for 12 h.Flow cytometry was used to detect apoptosis,and real-time PCR was used to measure the mRNA expression levels of nuclear factor kappa B(NF-κB),nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3(NLRP3),and interleukin-18(IL-18).Western blotting was used to detect the protein expression levels of NF-κB,NLRP3,and IL-18.Results The survival rates of myocardial cells were(53.32±5.70)%,(68.24±6.41)%,(92.62±8.56)%,and(86.93±7.12)%,respectively.The 20%drug-containing serum group was selected as the optimal concentration for drug intervention.The apoptosis rates of cardiomyocytes in the control group,model group,and SYYQ group were(1.57±0.18)%,(18.99±1.32)%,and(6.76±0.38)%,respectively.Compared with the control group,the apoptosis rate in the model group was significantly higher(P<0.01).Compared with the model group,the apoptosis rate in the SYYQ group was significantly lower(P<0.01).The mRNA expression levels of NF-κB,NLRP3,and IL-18 were significantly higher in the model group compared with the control group(P<0.05).Compared with the model group,the mRNA expression levels of NF-κB,NLRP3,and IL-18 were significantly lower in the SYYQ group(P<0.01).The relative protein expression levels of NLRP3 and IL-18

关 键 词：参元益气活血胶囊 心肌细胞 肿瘤坏死因子Α 炎症损伤 核因子κB 核苷酸结合寡聚域样受体蛋白3 白细胞介素18 小鼠 

分 类 号：R285.5[医药卫生—中药学]