冠通方含药血清通过miR-223/NLRP3信号通路调控血管内皮细胞焦亡的作用及机制研究  

Effect and Mechanism of Guantong Decoction-Containing Serum in Regulating Vascular Endothelial Cell Pyroptosis through miR-223/NLRP3 Signaling Pathway

作　　者：祝英杰 王强[1] 韦斌 尹玮[1] 李冬杏 谷林峰 ZHU Yingjie;WANG Qiang;WEI Bin;YIN Wei;LI Dongxing;GU Linfeng(Guangxi International Zhuang Medicine Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530000,Guangxi,China)

机构地区：[1]广西中医药大学附属国际壮医医院,南宁530000 [2]广西中医药大学附属瑞康医院 [3]广西中医药大学

出　　处：《中西医结合心脑血管病杂志》2023年第6期1037-1041,共5页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：国家自然科学基金项目(No.81960867)。

摘　　要：目的:探究冠通方含药血清通过microRNA-223(miR-223)/NOD样受体家族3(NLRP3)信号通路调控血管内皮细胞焦亡的作用及机制研究。方法:将血管内皮细胞系人脐静脉血管内皮细胞(HUVECs)分为对照组、模型组、空白血清组和含药血清组。对照组仅进行常规培养,其余3组采用氧化型低密度脂蛋白(ox-LDL)诱导HUVECs细胞损伤模型,模型组不进行任何干预,空白血清组采用空白血清干预,含药血清组则采用冠通方含药血清进行干预。实时荧光定量聚合酶链式反应(qRT-PCR)检测细胞中miR-223 mRNA表达,蛋白质印迹法(Western Blot)检测NLRP3、含半胱氨酸的天冬氨酸蛋白水解酶(Caspase1)的蛋白表达,酶联免疫吸附实验(ELISA)检测白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)的含量,细胞计数试剂盒(CCK8)检测细胞活性,并计算乳酸脱氢酶(LDH)释放水平。碘化丙啶(PI)染色荧光显微镜下观察细胞膜完整性和细胞焦亡情况。结果:与对照组相比,模型组HUVECs细胞活力明显降低(P<0.05);与模型组相比,空白血清组细胞活力差异无统计学意义(P>0.05),含药血清组的细胞活力则明显增强(P<0.05)。与对照组相比,模型组LDH释放水平明显升高(P<0.05);与模型组相比,空白血清组LDH释放水平差异无统计学意义(P>0.05),含药血清组LDH释放水平则明显下降(P<0.05)。与对照组相比,模型组PI染色程度明显增加;与模型组相比,空白血清组PI染色程度变化不明显,含药血清组PI染色程度明显减少。与对照组相比,模型组炎性因子IL-1β、IL-18含量明显升高(P<0.05);与模型组相比,空白血清组IL-1β、IL-18含量差异无统计学意义(P>0.05),含药血清组IL-1β、IL-18含量明显下降(P<0.05)。与对照组相比,模型组Caspase1蛋白表达明显上调;与模型组相比,空白血清组Caspase1蛋白表达差异无统计学意义(P>0.05),含药血清组Caspase1蛋白表达明显下调。与空白血清组相比,含药血�Objective:To explore the effect and mechanism of Guantong Decoction-containing serum in regulating the pyroptosis of vascular endothelial cells through microRNA-223(miR-223)/NOD-like receptor family 3(NLRP3)signaling pathway.Methods:Human umbilical vein endothelial cells(HUVECs)of vascular endothelial cell line were divided into control group,model group,blank serum group,and drug-containing serum group.The control group received routine culture,and the remaining three groups were treated with oxidized low density lipoprotein(ox-LDL)to induce HUVECs cell injury model,and the model group did not undergo any intervention,the blank serum group was intervened with blank serum,and the drug-containing serum group was treated with Guantong Decoction-containing serum.Quantitative real-time PCR(qRT-PCR)was used to detect the mRNA expression of miR-223 in cells,and Western Blot was applied to measure the protein expressions of NLRP3 and cysteinyl aspartate specific proteinase 1(Caspase1).Enzyme linked immunosorbent assay(ELISA)was adopted to detect the levels of interleukin-1β(IL-1β)and interleukin-18(IL-18),and cell counting kit-8(CCK8)was used to detect cell viability,and the release level of lactate dehydrogenase(LDH)was calculated.The cell membrane integrity and cell pyroptosis were observed under fluorescence microscopy with propidium iodide(PI)staining.Results:The HUVECs cell viability was significantly reduced in model group compared with that in control group(P<0.05),and the cell viability in blank serum group hardly changed compared with that in model group(P>0.05),but it was significantly enhanced in drug-containing serum group(P<0.05).In addition,compared with control group,the LDH release level of model group was significantly increased(P<0.05),while compared with model group,the LDH release level of blank serum group showed no significant change(P>0.05),and the LDH release level of drug-containing serum group was significantly decreased(P<0.05).The PI staining degree was significantly increased in model grou

关 键 词：血管内皮细胞 细胞焦亡 冠通方 miR-223 NOD样受体家族3 实验研究 

分 类 号：R285[医药卫生—中药学]