灯盏花乙素上调细胞外信号调节激酶表达拮抗人心脏微血管内皮细胞IR损伤

发布时间：2018-03-31 04:10

  本文选题：灯盏花乙素　切入点：缺血再灌注损伤　出处：《医学研究生学报》2017年09期

【摘要】：目的灯盏花乙素(SCU)可拮抗心肌IR损伤,但SCU是否通过细胞外信号调节激酶(ERK1/2)通路拮抗IR所致血管内皮细胞损伤尚不清楚。文中探讨SCU在缺氧-复氧(HR)诱导的人心脏微血管内皮细胞(HCMEC)损伤中的作用及对ERK1/2信号通路的影响。方法体外培养的HCMEC分别进行正常培养和缺氧12 h-复氧12 h建立IR模型(HR)处理。正常培养条件下,将HCMEC分为正常对照组、DMSO组、SCU 1μmol/L组和SCU 10μmol/L组。给予HCMEC IR损伤处理后,分为空白对照组、HR模型组、HR+DMSO组、HR+SCU 1μmol/L组和HR+SCU 10μmol/L组。将HCMEC与SCU或DMSO预孵育2 h后行HR处理。采用MTT法和锥虫蓝染色检测细胞存活率,并检测HCMEC的丙二醛(MDA)浓度。Western blot检测p-ERK1/2、ERK2和GAPDH蛋白表达情况。结果 MTT检测结果显示,与正常对照组细胞存活率(100%)比较,SCU 1μmol/L组和SCU 10μmol/L组[(110.40±2.34)%和(122±1.25)%]均增加(P0.05);与空白对照组细胞活力(100%)比较,HR模型组[(68.00±4.06)%]下降(P0.05),与HR模型组细胞存活率比较,HR+SCU 1μmol/L组和HR+SCU 10μmol/L组[(90.53±3.67)%、(92.04±2.32)%]增加(P0.05)。锥虫蓝染色显示,与正常对照组细胞活力[(90.06±1.85)%]比较,SCU 10μmol/L组[(96.78±2.01)%]增加(P0.05);与空白对照组[(91.83±2.34)%]比较,HR模型组细胞存活率[(73.72±4.91)%]下降(P0.01),与HR模型组比较,HR+SCU 10μmol/L组细胞存活率[(87.59±2.64)%]增加(P0.05)。与空白对照组比较,HR模型组、HR+DMSO组MDA明显增加(P0.01);与HR模型组比较,HR+SCU 1μmol/L组和HR+SCU 10μmol/L组MDA含量减少(P0.05)。与空白对照组比较,HR模型组p-ERK1/2蛋白表达明显降低(P0.01);与HR模型组比较,HR+SCU 10μmol/L组p-ERK1/2蛋白表达明显增加(P0.01)。结论 HR损伤导致HCMEC细胞活力下降、MDA增加和p-ERK1/2蛋白表达降低,SCU通过上调p-ERK1/2表达而拮抗HCMEC IR损伤。
[Abstract]:Objective to antagonize myocardial IR injury by Erigeron breviscapine (SCU). However, it is not clear whether SCU antagonizes vascular endothelial cell damage induced by IR through the extracellular signal-regulated kinase ERK1 / 2 pathway. In this paper, we investigate the role of SCU in human heart microvascular endothelial cell injury induced by hypoxia and reoxygenation. Methods HCMEC cultured in vitro was treated with normal culture and 12 h hypoxia and 12 h reoxygenation to establish IR model. HCMEC was divided into normal control group (1 渭 mol/L group) and SCU group (10 渭 mol/L group). HR SCU 1 渭 mol/L group and HR SCU 10 渭 mol/L group were divided into two groups. HCMEC was preincubated with SCU or DMSO for 2 hours. Cell survival rate was detected by MTT method and trypanosome blue staining. The malondialdehyde (MDA) concentration of HCMEC. Western blot was used to detect the expression of p-ERK1 / 2 ERK2 and GAPDH protein. Compared with the normal control group, the cell survival rate of SCU 1 渭 mol/L group and SCU 10 渭 mol/L group [110.40 卤2.34% and 122 卤1.25%] increased P0.05%, compared with the blank control group, the cell viability of HR model group [68.00 卤4.06%] decreased P0.05%, and the cell survival rate of HR SCU 1 渭 mol/L group and HR SCU 10 渭 mol/L group was higher than that of HR model group. [90.53 卤3.67] increased by 92.04 卤2.32%. Trypanosoma blue staining showed, Compared with the normal control group [90.06 卤1.85%], the cell viability of the SCU 10 渭 mol/L group [96.78 卤2.01U%] increased P0.05%, compared with the blank control group [91.83 卤2.34%], the cell viability of the HR model group [73.72 卤4.91%] decreased P0.01U, and the cell survival rate of the HR SCU 10 渭 mol/L group [87.59 卤2.641g] increased than that of the blank control group [87.59 卤2.641U], and increased the cell viability of the HR SCU 10 渭 mol/L group (87.59 卤2.641g%), compared with the control group (91.83 卤2.34g%) and the HR model group (93.72 卤4.91g%). Compared with HR model group, HR SCU 1 渭 mol/L group and HR SCU 10 渭 mol/L group decreased MDA content in HR SCU 1 渭 mol/L group and HR SCU 10 渭 mol/L group. Compared with blank control group, p-ERK1/2 protein expression in HR model group decreased significantly (P 0.01), and that in HR SCU 10 渭 mol/L group was significantly lower than that in HR model group, and that in HR SCU 10 渭 mol/L group was significantly lower than that in HR model group. Conclusion the activity of HCMEC cells decreased after HR injury and the expression of p-ERK1/2 protein decreased. Sch U antagonized HCMEC IR damage by upregulating the expression of p-ERK1/2.
【作者单位】： 昆明医科大学第一附属医院心内科;昆明医科大学病理教研室;昆明医科大学云南省天然药物药理重点实验室;
【基金】：国家自然科学基金(81560072) 云南省科学技术厅-昆明医科大学应用基础研究联合专项资金(2014FB037)
【分类号】：R54
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