盐酸戊乙奎醚对大鼠心肌微血管内皮细胞缺氧/复氧后的保护作用
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Effects of penehyclidine hydrochloride on Cardiac microvascular endotheliocyte apoptosis after hypoxia/reoxygenation injury in rats
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    摘要:

    目的:从细胞水平建立大鼠心肌微血管内皮细胞缺氧/复氧模型,观察了盐酸戊乙奎醚对心肌微血管内皮细胞的H/R损伤的保护作用。方法:实验采用酶消化法进行细胞的原代培养,1:2传代。采用荧光标记乙酰化Dil-Ac-LDL的方法鉴定。培养的细胞随机分为4组,①对照组,常氧环境下培养,不给予任何处理;②H/R组,改培养液为D-Hank,s液,于37℃、5%CO2、95%N2 密闭缺氧鑵内培养2h后,弃D-Hank,s液,改为完全培养液,放入常氧培养箱中继续培养4h;③PHC预处理组,在细胞缺氧2h前给予PHC(终浓度为0.1μm•L-1)预处理,④H/R +PHC组,在细胞H/R后,给予PHC(终浓度为0.1μm•L-1)孵育2h。MTT自动比色法测细胞活力,流式细胞法检测细胞凋亡。结果:①成功培养大鼠心肌微血管内皮细胞,用荧光标记乙酰化Dil-Ac-LDL的方法鉴定为微血管内皮细胞。②成功制备出微血管内皮细胞的H/R模型 将试验细胞培养液改为D-Hank,s 液,置于37℃、5%CO2、92%N2 密闭缺氧罐中环缺氧2h,后改为完全培养液放入常氧培养箱中继续培养4h可制备出微血管内皮细胞的I/R模型。③. H/R组细胞死亡率和凋亡率与对照组相比明显增高,差异有统计学意义(P<0.01),PHC+H/R组的细胞死亡率和凋亡率下降,差异有统计学意义(P<0.05),H/R+PHC组与对照组相比,虽然细胞的死亡率和凋亡率有所下降,但差异没有统计学意义(P>0.05)。结论:PHC预处理对大鼠微血管内皮细胞的H/RI有保护作用。 关键词:盐酸戊乙奎醚;心肌微血管内皮细胞;缺氧/复氧

    Abstract:

    Abstract: Objective Using the H/R model at cell leve, to observe protective effect and possible mechanism of penehyclidine Hydrochloride on myocardial I/R injur of microvascular endothelial cells.Methods In this experiment carry out cardiac vascular endothelial cells for primary culture were obtained by enzyme digestion1:2 passage.Fluorescent labeling acetylate Dil-Ac-LDL method was used to identification. Cutured cells were randomly divided into 4 groups, ① the normal control group, cultured under normoxic environment and did not give any treatment; ② H/R group,exchanged the culture medium into D-Hank,s liquid,putted it into 37℃、5%CO2、95%N2 a close tightly jar which was hypoxia for 2h,and then retrieved the total culture medium ,putted it into a normoxic environment to continue to culture for 4h; ③ PHC preconditon group,gived PHC(final concentration is 0.1μmol/L) hypoxia before; ④ H/R+ PHC group,after the cells H/R,gived PHC(final concentration is 0.1μmol/L),and continued to culture 2h.Results .The cardiac microvascular endothelial cells of adult rat were cultured successfully. Use fluorescent labeling acetylate Dil-Ac-LDL method to identify the cells. The H/R model of cardiac microvascular endothelial cells was established successfully. Exchange total culture medium into D-Hank,s liquid,put it into a close tightly jar which is on the condition of37℃、5%CO2、95%N2 hypoxia for 2h,and then retrieve the total culture medium ,put it into a normoxic environment to continue to culture for 4h. H/R group, the rate of cell death and apoptosis compared with the control group significantly increased, the difference was statistically significant(P<0.01); PHC+ H/R group,the cells death and apoptosis rate were declined than the H/R group, the differences were significance (P <0.05); H/R + PHC group compared with the control group, although the cells and the apoptosis rate of were decline, but the difference was not statistically significant(P>0.05).Conclusion PHC precondition has protective effects on cardiac microvascular endothelial cells after H/R injury in rats.

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兑宏志. 盐酸戊乙奎醚对大鼠心肌微血管内皮细胞缺氧/复氧后的保护作用[J]. 科学技术与工程, 2009, 9(21): .
Dui Hongzhi. Effects of penehyclidine hydrochloride on Cardiac microvascular endotheliocyte apoptosis after hypoxia/reoxygenation injury in rats[J]. Science Technology and Engineering,2009,9(21).

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  • 收稿日期:2009-07-15
  • 最后修改日期:2009-07-24
  • 录用日期:2009-07-28
  • 在线发布日期: 2009-11-02
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