葱白提取物对H_(2)O_(2)处理脑微血管内皮细胞活性、凋亡及氧化应激的影响  

Effects of Scallion Extract on Activity,Apoptosis and Oxidative Stress of Brain Microvascular Endothelial Cells Treated with H_(2)O_(2)

作　　者：齐献忠[1] 邢英瀛 秦慧兵[1] QI Xianzhong;XING Yingying;QIN Huibing

机构地区：[1]南阳市中心医院神经内科,河南南阳473000

出　　处：《新中医》2022年第1期15-20,共6页New Chinese Medicine

摘　　要：目的:探讨葱白提取物对过氧化氢(H_(2)O_(2))处理脑微血管内皮细胞活性、凋亡、氧化应激的影响及其可能作用机制。方法:体外培养大鼠RBMEC,H_(2)O_(2)处理大鼠脑微血管内皮细胞(RBMEC)建立细胞损伤模型,加入不同剂量的葱白提取物处理细胞,分别将pc DNA、pc DNA-CAI2转染至RBMEC,加入葱白提取物与H_(2)O_(2)共同处理细胞;采用甲基噻唑基四唑(MTT)法、流式细胞术分别检测细胞活性及凋亡率;采用实时荧光定量聚合酶链反应(q RT-PCR)法检测长链非编码RNA CAI2(Lnc RNA CAI2)的表达量;采用2,4-二硝基苯肼显色法检测乳酸脱氢酶(LDH)的活性;采用黄嘌呤氧化酶法检测超氧化物歧化酶(SOD)的活性;蛋白免疫印迹(Western blot)法检测Cleaved-caspase3、Bax、Pro-caspase3、Bcl-2蛋白表达量。结果:与Con组比较,H_(2)O_(2)处理后可明显降低细胞活性、SOD的活性及Pro-caspase3、Bcl-2蛋白水平(P<0.05),提高CAI2的表达水平、凋亡率、LDH的活性及Cleaved-caspase3、Bax蛋白水平(P<0.05),葱白提取物可明显逆转H_(2)O_(2)对RBMEC活性、凋亡率、氧化应激及CAI2表达量的作用;与H_(2)O_(2)+葱白-高+pc DNA组比较,H_(2)O_(2)+葱白-高+pc DNA-CAI2组细胞活性、SOD的活性及Pro-caspase3、Bcl-2蛋白水平降低(P<0.05),凋亡率、LDH的活性及Cleaved-caspase3、Bax蛋白水平显著升高(P<0.05)。结论:葱白提取物可通过抑制CAI2的表达而增强H_(2)O_(2)诱导的RBMEC活性及抑制细胞凋亡、氧化应激,进而对RBMEC细胞损伤发挥保护作用。Objective:To investigate the effects of scallion extract on the activity,apoptosis and oxidative stress of brain microvascular endothelial cells treated with hydrogen peroxide(H_(2)O_(2))and its possible mechanism.Methods:Rat brain microvascular endothelial cells(RBMEC)were cultured in vitro.RBMEC was treated with H_(2)O_(2) to establish the cell damage model.Different doses of scallion extract were added to treat the cells.pc DNA and pc DNA-CAI2 were transfected into RBMEC respectively.Scallion extract and H_(2)O_(2) were added to treat the cells together;MTT assay and flow cytometry were used to detect cell activity and apoptosis rate.The expression of long chain noncoding RNA CAI2(Lnc RNA CAI2)was detected by real-time fluorescence quantitative polymerase chain reaction(q RT-PCR).The activity of LDH was detected by 2,4-dinitrophenylhydrazine colorimetry.The activity of SOD was detected by xanthine oxidase method.The expressions of Cleaved-caspase3,Bax,Pro-caspase3 and Bcl-2 were detected by Western blot.Results:Compared with con group,H_(2)O_(2) treatment could significantly reduce cell activity,SOD activity,Pro-caspase3 and Bcl-2 protein levels(P<0.05),increase CAI2 expression level,apoptosis rate,LDH activity and Cleaved-caspase3 and Bax protein levels(P<0.05).Scallion extract could significantly reverse the effects of H_(2)O_(2) on RBMEC activity,apoptosis rate,oxidative stress and CAI2expression.Compared with H_(2)O_(2)+scallion high+pc DNA group,the cell activity,SOD activity and Pro-caspase3 and Bcl-2protein levels in H_(2)O_(2)+scallion high+pc DNA-CAI2 group decreased(P<0.05),and the apoptosis rate,LDH activity and Cleaved-caspase3 and Bax protein levels increased significantly(P<0.05).Conclusion:Scallion extract can enhance the activity of RBMEC induced by H_(2)O_(2),inhibit cell apoptosis and oxidative stress,and then protect RBMEC cells from damage by inhibiting the expression of CAI2.

关 键 词：葱白提取物 过氧化氢 脑微血管内皮细胞 LncRNA CAI2 细胞活性 凋亡 氧化应激 

分 类 号：R285.5[医药卫生—中药学]