肾衰康灌肠液含药血清对HK-2细胞缺氧/复氧损伤的保护作用及对Caspase-3、Bax、Bcl-2mRNA表达的影响  

Protective Effect of Shenshuaikang Enema(肾衰康灌肠液)-Containing Serum on Hypoxia/Reoxygenation Injury of HK-2 Cells and Its Effect on Caspase-3,Bax and Bcl-2 mRNA

作　　者：陆红梅 何玉华[1] 罗芯怡[1] 叶乃菁 瞿波 李明权[1] LU Hongmei;HE Yuhua;LUO Xinyi;YE Naijing;QU Bo;LI Mingquan(Hospital of Chendu University of Traditional Chinese Medicine,Chengdlu 610075,Sichuan,China)

机构地区：[1]成都中医药大学附属医院,四川成都610075

出　　处：《中华中医药学刊》2021年第3期218-222,I0034-I0038,共10页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81973831)。

摘　　要：目的观察肾衰康灌肠液含药血清对缺氧/复氧损伤肾小管上皮细胞(HK-2细胞)凋亡的抑制作用,并探讨其对缺氧/复氧损伤HK-2细胞凋亡相关基因Caspase-3、Bax、Bcl-2表达的影响。方法将家兔随机分为空白组、PBS组、肾衰康组,予相应灌肠3 d后取血清,分别制备含空白血清、PBS血清、25%肾衰康灌肠液含药血清、50%肾衰康灌肠液含药血清、75%肾衰康灌肠液含药血清,利用各组血清分别处理缺氧/复氧损伤HK-2细胞。利用ROS荧光探针检测缺氧/复氧损伤情况,利用CFSE/PI、流式细胞术检测细胞凋亡情况;利用RT-PCR检测各组HK-2细胞Caspase-3、Bax、Bcl-2基因表达情况。结果与造模前相比,造模后细胞内ROS荧光强度增强,提示造模成功。CFSF/PI结果显示,与模型组相比、PBS组,肾衰康各组细胞凋亡明显被抑制,呈剂量依赖性。流式细胞术结果显示,与对照组[4 h,(42.7±3.5)%;12 h,(41.9±2.2)%]、PBS组[4 h,(40.9±1.2)%;12 h,(39.8±0.7)%]比较,肾衰康各组凋亡/死亡率明显被抑制(P<0.05),低剂量组[4 h,(18.5±0.2)%;12 h,(17.3±0.3)%]、中剂量组[4 h,(13.2±2.1)%;12 h,(14.1±0.6)%]、高剂量组[4 h,(10.7±0.1)%;12 h,(12.2±0.9)%]。RT-PCR结果显示,与对照组、PBS组比较,肾衰康各组Caspase-3、Bax mRNA表达明显被抑制(P<0.05),Bcl-2mRNA表达明显增加(P<0.05),以高剂量组明显。结论肾衰康灌肠液可以有效改善HK-2细胞缺氧/复氧损伤作用,其机制与下调促凋亡基因Caspase-3、Bax表达,上调抗凋亡基因Bcl-2表达有关。Objective To observe the inhibitory effect of Shenshuaikang Enema(肾衰康灌肠液)-containing serum on the renal tubular epithelial cell(HK-2)injured by hypoxia/reoxygenation injury and explore its effect on the expressions of apoptosis-related gene Caspase-3,Bax and Bcl-2 mRNA.Methods The rabbits were randomly divided into blank group,PBS group and Shenshuaikang Enema group,and the serum was taken out 3 days after the corresponding enema.The blank serum,PBS serum,25%Shenshuaikang Enema-containing serum and 50%Shenshuaikang Enema-containing serum,75%Shenshuaikang Enema-containing serum were prepared.The above containing serum was used to treat hypoxia/reoxygenation injury HK-2 cells.ROS fluorescence probes were used to detect hypoxia/reoxygenation injury,CFSE/PI and flow cytometry was used to detect apoptosis and RT-PCR was used to detect expressions of Caspase-3,Bax and Bcl-2 mRNA in HK-2 cells of each group.Results Compared with those before modeling,the intracellular ROS fluorescence intensity was enhanced after modeling,suggesting that the modeling was successful.The CFSF/PI results showed compared with that of the model group,the apoptosis of the Shenshuaikang Enema group was significantly inhibited in a dose-dependent manner.Flow cytometry results showed compared with those of the control group[4 h,(42.7±3.5)%;12 h,(41.9±2.2)%]and the PBS group[4 h,(40.9±1.2)%;12 h,(39.8±0.7)%],the apoptosis/mortality of the Shenshuaikang Enema group was significantly inhibited(P<0.05).The apoptosis/mortality of the low dose group was 4 h[(18.5±0.2)%]and 12 h[(17.3±0.3)%].The apoptosis/mortality of the medium dose group was 4 h[(13.2±2.1)%]and 12 h[(14.1±0.6)%].The apoptosis/mortality of high-dose group was 4 h[(10.7±0.1)%]and 12 h[(12.2±0.9)%].The results of RT-PCR showed that compared with those of the control group and the PBS group,the expressions of Caspase-3 and Bax mRNA were significantly inhibited(P<0.05),and the expression of Bcl-2 mRNA was significantly increased(P<0.05),and those of the high dose gro

关 键 词：肾衰康灌肠液 细胞凋亡 CASPASE-3 BAX bal-2 

分 类 号：R285.5[医药卫生—中药学]