葱白提取物对H2O2处理脑微血管内皮细胞活性、凋亡及氧化应激的影响
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R285.5

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Effects of Scallion Extract on Activity, Apoptosis and Oxidative Stress of Brain Microvascular Endothelial Cells Treated with H2O2
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    摘要:

    目的:探讨葱白提取物对过氧化氢(H2O2) 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,H2O2 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pcDNA、pcDNA-CAI2 转染至RBMEC,加入葱白提取物与H2O2 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(qRT-PCR) 法检测长链非编码RNACAI2(LncRNA 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,H2O2 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H2O2 对RBMEC 活性、凋亡率、氧化应激及CAI2 表达量的作用;与H2O2+葱白-高+pcDNA 组比较,H2O2+葱白-高+pcDNA-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H2O2 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。

    Abstract:

    Abstract:Objective:To investigate the effects of scallion extract on the activity,apoptosis and oxidative stress of brain microvascular endothelial cells treated with hydrogen peroxide(H2O2) and its possible mechanism. Methods: Rat brain microvascular endothelial cells(RBMEC) were cultured in vitro. RBMEC was treated with H2O2 to establish the cell damage model. Different doses of scallion extract were added to treat the cells. pcDNA and pcDNA- CAI2 were transfected into RBMEC respectively. Scallion extract and H2O2 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(LncRNA CAI2) was detected by real- time fluorescence quantitative polymerase chain reaction(qRT- 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, H2O2 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 H2O2 on RBMEC activity, apoptosis rate, oxidative stress and CAI2 expression.Compared with H2O2 + scallion high + pcDNA group,the cell activity,SOD activity and Pro- caspase3 and Bcl- 2 protein levels in H2O2 + scallion high + pcDNA- 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 H2O2,inhibit cell apoptosis and oxidative stress,and then protect RBMEC cells from damage by inhibiting the expression of CAI2.

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齐献忠,邢英瀛,秦慧兵.葱白提取物对H2O2处理脑微血管内皮细胞活性、凋亡及氧化应激的影响[J].新中医,2022,54(1):15-20

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  • 在线发布日期: 2022-01-10
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