香青兰总黄酮对缺氧缺糖/复氧损伤和内质网应激诱导H9c2细胞凋亡的影响  

Effects of total flavonoids of Dracocephalum moldavica on apoptosis of H9c2 cells induced by OGD/R injury and endoplasmic reticulum stress

作　　者：王天 刘砥威 王桐叶 张兴宇 邢建国 郑瑞芳 WANG Tian;LIU Di-wei;WANG Tong-ye;ZHANG Xing-yu;XING Jian-guo;ZHENG Rui-fang(Xinjiang Medical University,Urumqi 830011,China;Xinjiang Institute of Pharmacy,Urumqi 830002,China;Shihezi University,Shihezi832000,China)

机构地区：[1]新疆医科大学,新疆乌鲁木齐830011 [2]新疆药物研究所,新疆乌鲁木齐830002 [3]石河子大学,新疆石河子832000

出　　处：《中国中药杂志》2025年第5期1321-1330,共10页China Journal of Chinese Materia Medica

基　　金：新疆维吾尔自治区自然科学基金项目(2022D01A299,2022D01D50);新疆维吾尔自治区“天山英才”科技创新领军人才项目(2023TSYCLJ0042);新疆维吾尔自治区“天山英才”青年托举人才项目(2023TSYCQNTJ0010)。

摘　　要：探讨香青兰总黄酮(total flavonoids of Dracocephalum moldavica,TFDM)对大鼠H9c2细胞通过缺氧缺糖/复氧(oxygen-glucose deprivation and reoxygenation,OGD/R)损伤和衣霉素(tunicamycin,TM)建立内质网应激(endoplasmic reticulum stress,ERS)诱导细胞凋亡的影响,并探索可能的作用机制。造模成功后,将大鼠分为对照组,模型(OGD/R或TM)组,TFDM低、中、高(12.5、25、50μg·mL^(-1))剂量组。体外构建OGD/R损伤模型,cell counting kit-8(CCK-8)法检测TFDM对细胞增殖的影响;检测细胞上清液乳酸脱氢酶(lactate dehydrogenase,LDH)和肌酸激酶同工酶MB(creatine kinase MB isoenzyme,CKMB)水平;Western blot检测葡萄糖调节蛋白78(glucose regulatory protein 78,GRP78)、C/EBP同源蛋白(C/EBP homologous protein,CHOP)、活化转录因子6(activating transcription factor 6,ATF6)和B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X-protein,Bax)的表达。采用脱氧核苷酸末端转移酶介导的dUTP缺口末端标记(terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling,TUNEL)法检测细胞凋亡。TM诱导ERS模型中,Western blot检测TM组中ERS通路相关蛋白GRP78、CHOP、肌醇需求酶1(inositol-requiring enzyme 1,IRE1)、X-框结合蛋白1(X-box binding protein 1,XBP1)、蛋白激酶RNA样内质网激酶(protein kinase RNA-like endoplasmic reticulum kinase,PERK)、真核翻译起始因子2α(eukaryotic initiation factor 2α,eIF2α)、ATF6、磷酸化ATF6(phospho-ATF6,p-ATF6)和凋亡相关蛋白Bcl-2、Bax、半胱天冬氨酸蛋白水解酶-12(cysteinyl aspartate specific proteinase-12,caspase-12)、活性caspase-12(cleaved caspase-12,cleaved caspase-12)的表达;实时荧光定量PCR(real-time fluorescence quantitative PCR,RT-qPCR)检测GRP78、CHOP、PERK和ATF6的基因表达;采用TUNEL法检测细胞凋亡情况。结果显示,在OGD/R模型中,与对照组相比,OGD/R组细胞上清液LDH和CKMB水平显著升高;与OGD/R组相比,TFDM组细胞上清液LDH和CKMB水平显著This study investigated the effects of total flavonoids of Dracocephalum moldavica(TFDM)on apoptosis in rat H9c2 cells induced by endoplasmic reticulum stress(ERS)established by oxygen-glucose deprivation and reoxygenation(OGD/R)injury and tunicamycin(TM),and explored the potential mechanisms.After successful modeling,the following groups were set in this experiment:control group,model(OGD/R or TM)group,and TFDM low-,medium-,and high-dose groups(12.5,25,and 50μg·mL^(-1)).The OGD/R injury model was constructed in vitro.Cell proliferation was assessed using the cell counting kit-8(CCK-8)method.The levels of lactate dehydrogenase(LDH)and creatine kinase MB isoenzyme(CKMB)in the cell supernatant were detected.Western blot was used to assess the expression of ERS-related proteins,including glucose regulatory protein 78(GRP78),C/EBP homologous protein(CHOP),activating transcription factor 6(ATF6),and apoptotic proteins B-cell lymphoma 2(Bcl-2)and Bcl-2-associated X protein(Bax).Apoptosis was detected using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)method.In the TM-induced ERS model,Western blot was used to measure the expression of ERS pathway-related proteins GRP78,CHOP,inositol-requiring enzyme 1(IRE1),X-box binding protein 1(XBP1),protein kinase RNA-like endoplasmic reticulum kinase(PERK),eukaryotic initiation factor 2α(eIF2α),ATF6,p-ATF6,and apoptotic proteins Bcl-2,Bax,cysteinyl aspartate specific proteinase-12(caspase-12),and cleaved caspase-12.Gene expression of GRP78,CHOP,PERK,and ATF6 was detected by real-time fluorescence quantitative PCR(RT-qPCR).Apoptosis was again detected using the TUNEL method.The results showed that in the OGD/R model,compared with the control group,the levels of LDH and CKMB in the cell supernatant were significantly increased in the OGD/R group.Compared with the OGD/R group,the levels of LDH and CKMB in the TFDM group were significantly reduced.Western blot results revealed that compared with the control group,the expression of ERS-related pro

关 键 词：香青兰总黄酮 H9C2细胞 细胞凋亡 衣霉素 内质网应激 缺氧缺糖/复氧 

分 类 号：R285.5[医药卫生—中药学]