马齿苋总黄酮通过调控TRB3表达对支气管上皮细胞缺氧/复氧损伤的影响  

Effects of Portulaca oleracea total flavones on hypoxic/reoxygenation injury of bronchial epithelial cells by regulating TRB3 expression

作　　者：付敏 程弘夏 桑筱筱 FU Min;CHENG Hongxia;SANG Xiaoxiao(School of Biology and Pharmaceutical Engineering,Wuhan Huaxia University of Technology,Wuhan 430223,China)

机构地区：[1]武汉华夏理工学院生物与制药工程学院,湖北武汉430223

出　　处：《沈阳药科大学学报》2022年第12期1501-1508,共8页Journal of Shenyang Pharmaceutical University

基　　金：2018年湖北省科技厅计划项目(2018CFC888)。

摘　　要：目的探讨马齿苋总黄酮(PTF)是否通过调控tribbles同源蛋白3(tribbles homolog 3,TRB3)表达影响缺氧/复氧(H/R)诱导的支气管上皮细胞损伤。方法体外培养人支气管上皮细胞BEAS-2B,建立H/R诱导的细胞损伤模型,使用低(10 mg·L^(-1))、中(20 mg·L^(-1))、高浓度(40 mg·L^(-1))PTF处理细胞;用试剂盒检测乳酸脱氢酶(lactate dehydrogenase,LDH)、丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)的含量;流式细胞术检测细胞凋亡率;采用实时荧光定量PCR(qRT-PCR)与Western blot法分别检测TRB3的mRNA及蛋白表达水平;再将si-TRB3、pcDNA-TRB3分别转染至BEAS-2B细胞,采用上述方法检测转染后LDH、MDA、SOD的水平及细胞凋亡率;Western blot法检测B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、B淋巴细胞瘤-2相关蛋白(Bcl-2-associated X protein,Bax)蛋白表达量。结果与对照组比较,H/R组LDH、MDA的水平升高,SOD的水平降低,细胞凋亡率升高,Bax蛋白水平升高,Bcl-2蛋白水平降低,TRB3 mRNA及蛋白水平升高;与H/R组比较,H/R+PTF不同浓度组LDH、MDA的水平均降低,SOD的水平升高,细胞凋亡率降低,Bax蛋白水平降低,Bcl-2蛋白水平升高,TRB3 mRNA及蛋白水平降低;与H/R+si-NC组比较,H/R+si-TRB3组LDH、MDA的水平降低,SOD的水平升高,细胞凋亡率降低,Bax蛋白水平降低,Bcl-2蛋白水平升高;TRB3过表达可减弱PTF对H/R诱导的支气管上皮细胞损伤的作用。结论马齿苋总黄酮可能通过下调TRB3的表达从而减轻H/R诱导的支气管上皮细胞损伤。Objective To investigate whether total flavonoids(PTF)affect hypoxia/reoxygenation(H/R)-induced bronchial epithelial cell damage by regulating the expression of tribbles homologous protein 3(tribbles homolog 3,TRB3).Methods BEAS-2 B cells were cultured in vitro to establish the H/R-induced cell damage model,and cells were treated using low(10 mg·L^(-1)),medium(20 mg·L^(-1)),and high concentration(40 mg·L^(-1))PTF,and in subsequent experiments si-trb3,pcdna-trb3 were transfected into BEAS-2 B cells,respectively;The levels of lactate dehydrogenase(LDH),malondialdehyde(MDA),superoxide dismutase(SOD)were measured using kits;The apoptosis rate was detected by flow cytometry;TRB3 mRNA was quantified by real-time PCR(qRT-PCR);The protein expressions of TRB3,B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)were determined by Western blot analysis.Results Compared with the control group,the levels of LDH and MDA increased,the levels of SOD decreased,the apoptosis rate increased,the levels of Bax protein increased,the levels of Bcl-2 protein decreased,and the levels of TRB3 mRNA and protein increased in the H/R group.Compared with the H/R group,the levels of LDH and MDA were decreased,the levels of SOD were increased,the apoptosis rate was decreased,the levels of Bax protein were decreased,the levels of Bcl-2 protein were increased,and the levels of TRB3 mRNA and protein were decreased in the H/R+PTF group at different concentrations.Compared with the H/R+si-NC group,the H/R+si-TRB3 group showed decreased levels of LDH and MDA,increased levels of SOD,decreased apoptosis rate,decreased levels of Bax protein,and increased levels of Bcl-2 protein.TRB3 overexpression attenuated the effect of PTF on H/R-induced bronchial epithelial cell injury.Conclusion Portulaca oleracea total flavones may attenuate H/R-induced bronchial epithelial cell injury by downregulating TRB3 expression.

关 键 词：马齿苋总黄酮 TRB3 支气管上皮细胞 缺氧/复氧 凋亡 氧化应激 

分 类 号：R96[医药卫生—药理学]