补心软脉颗粒含药血清对AngⅡ诱导的HUVEC细胞抗氧化作用
Antioxidant Effects of Buxin Ruanmai Granules Serum on AngⅡ-induced Human Umbilical Vein Endothelial Cells
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摘要: 目的 观察补心软脉颗粒含药血清对AngⅡ诱导HUVEC细胞所致氧化损伤的相关指标SOD、NADPH、MDA、p40phox、p47phox、p67phox的影响。方法 大鼠被随机分为正常组、缬沙坦组、补心软脉颗粒组,每组10只,灌胃6周,取动脉血制备药物血清。建立10-6mol/L AngⅡ诱导的HUVEC细胞凋亡损伤模型,并予各药物血清干预,采用黄嘌呤氧化酶法检测细胞上清液中总SOD活力,ELISA法检测NADPH氧化酶,TBA法检测MDA含量,Bradford法p40phox、p47phox、p67phox蛋白定量;Western blot法分别检测细胞膜与细胞质p40phox、p47phox、p67phox蛋白表达。结果 AngⅡ诱导HUVEC细胞损伤,模型组SOD活力下降,NADPH、MDA升高,p40phox、p47phox、p67phox向细胞膜转移,出现氧化损伤;补心软脉颗粒组SOD活力上升,MDA、NADPH下降,抑制p40phox、p47phox、p67phox向细胞膜转移,减轻氧化损伤。结论 补心软脉含药血清对AngⅡ诱导体外培养的HUVEC损伤有抑制氧化作用,防止动脉硬化。Abstract: OBJECTIVE To observe the effects of Buxin Ruanmai granules (BXRM) serum on AngiotensinⅡ(AngⅡ)-induced oxidative damage related indicators, such as SOD,NADPH ,MDA,p40phox,p47phox and p67phox, in HUVEC. METHODS Rats were randomly divided into normal group,Valsartan group and BXRM group,each group of 10, and lavaged for 6 weeks. Drug serum were prepared from arterial blood. Then 10-6 mol/L Ang Ⅱ induced HUVEC damage and apoptosis model of human umbilical vein endothelial cells was established, and drug serum was used to intervene. SOD were determined by xanthinoxidase. NADPH was determined by ELISA. MDA was determined by thibabituric acid method. The expression of p40phox, p47phox and p67phox in cell membrane and cytoplasm of HUVEC were determined by Western blot. RESULTS After HUVECs were damaged by 10-6 mol/L Ang Ⅱ,the content of SOD decreased , NADPH and MDA increased , and p40phox, p47phox and p67phox transferred to the membrane of HUVECs. However, the content of SOD increased, NADPH and MDA decreased, and p40phox, p47phox and p67phox were inhibited to transfer to the membrane, after the medication of BXRM serum. CONCLUSION BXRM serum can inhibited the oxidation of AngⅡ induced HUVEC damage and prevent arteriosclerosis.
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Key words:
- Buxin Ruanmai granules /
- drug serum /
- AngⅡ /
- HUVEC /
- oxidation inhibition
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