健脾通络饮对缺氧再复神经细胞损伤保护作用机制的影响  

Effect of Jianpi Tongluo Decoction(健月卑通络饮)on the Protective Mechanism of Hypoxia Reactivated SH一SY5Y Injury

作　　者：陈枫 刘海兴[1] CHEN Feng;LIU Haixing(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning Province)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847

出　　处：《实用中医内科杂志》2021年第6期102-105,I0010,I0011,共6页Journal of Practical Traditional Chinese Internal Medicine

基　　金：辽宁省科技计划项目(2011408004)。

摘　　要：目的观察健脾通络饮对神经细胞(SH-SY5Y)缺氧损伤后是否有保护作用,探讨健脾通络饮防治脑缺血的作用机制。方法制备健脾通络饮含药血清,培养SH-SY5Y,将其分成正常对照组(C)、缺氧再复模型组(IR)、缺氧+健脾通络饮含药血清低浓度组(JL)、缺氧+健脾通络饮含药血清中浓度组(JM)、缺氧+健脾通络饮含药血清高浓度组(JH)、缺氧+银杏叶阳性对照组(GC),用低糖缺氧的DMEM培养基处理SH-SY5Y模拟细胞缺氧,Hoechst染色法检测神经细胞存活率;Western Blot检测细胞中Bcl-2和Bax蛋白的表达;流式细胞仪检测分析缺氧2 h处理后SH-SY5Y凋亡情况。结果Hoechst染色结果显示,给予中、高浓度的健脾通络饮含药血清处理的组别与缺氧/复氧损伤处理的模型组相比凋亡数量明显减少。WB结果显示,在含药血清的干预下,与正常对照组相比,模型组中Bax蛋白表达升高,Bcl-2/Bax比值明显下降(P<0.05);中、高浓度组与模型组相比,Bcl-2蛋白表达量升高,Bax下降,两者比值明显升高(P<0.05)。流式细胞仪检测结果,中、高浓度的健脾通络饮含药血清和银杏叶片阳性对照组与模型组相比,凋亡率下降,差异有统计学意义(P<0.05)。结论健脾通络饮可能对脑缺血再灌注脑损伤神经具有保护作用,其保护机制可能与其下调凋亡蛋白Bcl-2和Bax表达有关。Objective To investigate the protective effects of Jianpi Tongluo Decoction(健脾通络饮)on nerve cells(SH-SY5 Y) induced by hypoxia, and to study the mechanism of JianpiTongluo Decoction in preventing and treating cerebral ische-mia.MethodsPreparation of JianpiTongluo Yin drug-containing serum, Culture of SHSY-5 Y, divided into a normal controlgroup(C),hypoxia-reperfusion model group(IR), hypoxia + JianpiTongluo Yin low concentration group(JL), hypoxia +JianpiTongluo dection drug serum concentration group( JM), Hypoxia + JianpiTongluo Yin drug serum high concentrationgroup(JH), hypoxia + Ginkgo biloba positive control group(GC), SHSY-5 Y were treated with DMEM medium,which lowglucose and hypoxia to simulate cell hypoxia, and Hoechst staining was used to detect the survival rate of nerve cells;The pro-tein expression of Bcl-2 and Bax in cells was detected by Western Blot;Flow cytometry was used to detect the apoptosis ofnerve cells after 2 h hypoxia treatment.ResultsHoechst staining results showed, compared with the model group treated of an-oxia/reoxygenation injury, the number of apoptosis was significantly reduced in the group treated with Jianpi Tongluoyin con-taining drug serum at medium or high concentration. WB results showed that compared with the normal control group, the ex-pression of Bax protein in model group was increased and the Bcl-2/Bax ratio was significantly decreased(P< 0. 05) afterthe intervention of drug serum. Compared with model group, Bcl-2 protein expression was increased and Bax was decreasedin medium and high concentration groups, and the ratio of Bcl-2 protein expression and Bax protein expression was signifi-cantly increased(P< 0. 05). The results of flow cytometry showed that compared with the model group, the apoptosis rate ofthe positive control group of Jianpitongluoyin and Ginkgo biloba leaves with medium and high concentrations decreased, andthe difference was statistically significant(P< 0. 05).ConclusionJianpi Tongluo Decoction may have a protective effect on cer-ebral isc

关 键 词：健脾通络饮 人神经母细胞瘤细胞 含药血清 凋亡 

分 类 号：R285.5[医药卫生—中药学]