大黄醇提物对氯化血红素诱导HT22细胞自噬和凋亡的影响  

Effects of rhubarb ethanol extract on hemin-induced autophagy and apoptosis of HT22 cells

作　　者：岳孟茹 汪坤[2] 纪晓宁[2] 杨艳华 雷震[2] 马鸣 赵旭[2] YUE Meng-ru;WANG Kun;JI Xiao-ning;YANG Yan-hua;LEI Zhen;MA Ming;ZHAO Xu(School of Pharmacy,Henan University of Chinese Medicine,Zhengzhou 450046,China;TheSecond Hospital Affiliated to Henan University of Chinese Medicine,Zhengzhou 450002,China)

机构地区：[1]河南中医药大学药学院,河南郑州450046 [2]河南中医药大学第二附属医院,河南郑州450002

出　　处：《中成药》2023年第12期3930-3936,共7页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金面上项目(81573919);河南省中医药科学研究重大专项20-21ZYZD08;河南省中医药拔尖人才培养项目(豫卫中医函[2021]15号)。

摘　　要：目的探究大黄醇提物对氯化血红素诱导损伤后HT22细胞自噬与凋亡的影响。方法将HT22细胞分为正常组、模型组和大黄醇提物高、中、低剂量组(25、12.5、6.25μg/mL),给药干预24 h后,以40μmol/L氯化血红素诱导3 h建立细胞脑出血损伤模型。造模结束后,CCK-8法检测细胞存活率,试剂盒检测乳酸脱氢酶(LDH)漏出率,流式细胞术检测细胞凋亡率,Western blot法检测细胞凋亡及自噬相关蛋白半胱氨酸蛋白酶-3(caspase-3)、Bcl-2相关X蛋白(Bax)、B细胞淋巴瘤-2(Bcl-2)、微管相关蛋白轻链3(LC3)、Beclin1及P62表达。结果与正常组比较,模型组细胞存活率降低(P<0.01),LDH漏出率、细胞凋亡率和Bax/Bcl-2、caspase-3、Beclin1、LC3蛋白表达升高(P<0.01),P62蛋白表达降低(P<0.01);与模型组比较,大黄醇提物各剂量组细胞存活率升高(P<0.05,P<0.01),LDH漏出率、细胞凋亡率和Bax/Bcl-2、caspase-3、Beclin1、LC3蛋白表达降低(P<0.05,P<0.01),大黄醇提物高、中剂量组P62蛋白表达升高(P<0.05,P<0.01)。结论大黄醇提物可能通过调节自噬与凋亡减轻细胞损伤,实现对神经元的保护。AIM To explore the effects of rhubarb ethanol extract on autophagy and apoptosis of HT22 cells after hemin-induced injury.METHODS The HT22 cells were divided into the blank group,the model group,and the high,medium and low dose rhubarb ethanol extract groups(25,12.5 and 6.25μg/mL).After the 24 hours corresponding drug intervention,3 hours induction of 40μmol/L hemin was used for the modeling of cell cerebral hemorrhage injury.Subsequently,the cells had their cell viability detected by CCK-8 method;their leakage rate of lactate dehydrogenase(LDH)detected by kit;their apoptosis rate detected by flow cytometry;and their apoptosis and autophagy related protein expressions of cysteine protease-3(caspase-3),Bcl-2 related X protein(Bax),B cell lymphoma-2(Bcl-2)and microtubule related protein light chain 3(LC3)detected by Western blot.RESULTS Compared with the normal group,the model group displayed decreased cell viability(P<0.01);increased LDH leakage rate,apoptosis rate and the protein expressions of Bax/Bcl-2,caspase-3,Beclin1 and LC3(P<0.01);and decreased protein expression of P62(P<0.01).Compared with the model group,all of the groups intervened with rhubarb ethanol extract showed increased cell viability(P<0.05,P<0.01),decreased LDH leakage rate,apoptosis rate and protein expressions of Bax/Bcl-2,caspase-3,Beclin1 and LC3(P<0.05,P<0.01),in addition to the decreased protein expression of P62 increased(P<0.05,P<0.01)in the high and middle dose rhubarb ethanol extract groups.CONCLUSION The ethanol extract of rhubarb may protect neurons and reduce cell damage through its efficacy in regulating their autophagy and apoptosis.

关 键 词：大黄醇提物 HT22细胞 氯化血红素 脑出血 自噬 凋亡 

分 类 号：R285.5[医药卫生—中药学]