细颗粒物对人脐静脉内皮细胞ACE、AT1R基因表达的影响

发布时间：2018-06-20 06:57

本文选题：细颗粒物 + 血管内皮细胞　；参考：《中南大学》2012年硕士论文

【摘要】：目的：分析细颗粒物(PM2.5)对人脐静脉内皮细胞(HUVEC)活力和ACE、 ATlR mRNA和蛋白表达的影响,探讨培哚普利对HUVEC细胞ACE、 AT1R表达的调控。方法：于2010年10月-2011年12月采集长沙市非工业区大气的PM2.5,并提取PM2.5的水溶性和非水溶性成分。分别用100μg/ml、200μg/ml、400μg/ml(称为低浓度、中浓度、高浓度)PM2.5水溶性和非水溶性成分染毒HUVEC细胞24h。采用MTT法比较不同浓度PM2.5水溶性和非水溶性成分染毒对HUVEC细胞活力的影响,采用实时荧光定量PCR和Western Blot方法分析HUVEC细胞ACE、 ATlR mRNA和蛋白表达的差异。利用高浓度PM2.5染毒HUVEC细胞Oh、12h、24h和48h,比较不同时间PM2.5染毒对细胞活力和ACE, AT1R基因表达的影响。同时在PM2.5染毒HUVEC细胞时加入10μmol/L的培哚普利进行干预,分析培哚普利对HUVEC细胞ACE, AT1R表达的调控。结果： 1.PM2.5对HUVEC细胞活力的抑制效应：水溶性和非水溶性成分各浓度组细胞存活率均低于空白对照组(P0.05),且高浓度组中浓度组低浓度组(P0.05)。当染毒剂量同为中、高浓度时,非水溶性成分对细胞活力影响较水溶性成分明显(P0.05)。高浓度PM2.5染毒不同时间组细胞存活率均低于0h组(P0.05),且48h组24h组12h组(P0.05)。 2.PM2.5对HUVEC细胞ACE基因表达的影响：水溶性成分中、高浓度组和非水溶性成分各浓度组细胞ACEmRNA和蛋白表达量均高于空白对照组(P0.05),且非水溶性成分高浓度组ACE表达量最高(P0.05)。高浓度水溶性成分24h、48h组和非水溶性成分各不同时间组细胞ACE基因表达量高于Oh组(P0.05),且非水溶性成分24h组ACE表达量最高(P0.05)。 3.PM2.5对HUVEC细胞AT1R基因表达的影响：水溶性和非水溶性成分高浓度组细胞AT1RmRNA和蛋白表达量高于空白对照组(P0.05),且非水溶性成分高浓度组AT1R表达量最高(P0.05)。高浓度水溶性和非水溶性成分各不同时间组细胞AT1R表达量高于Oh组(P0.05),且非水溶性成分48h组AT1R表达量最高(P0.05)。 4.培哚普利干预对HUVEC细胞ACE、 AT1R表达的调控：培哚普利干预后,水溶性和非水溶性成分低、中、高浓度组细胞ACEmRNA和蛋白表达量均显著低于未加药物组(P0.05)；但AT1R mRNA和蛋白表达量无显著改变(P0.05)。结论： 1.PM2.5染毒可导致HUVEC细胞活力降低,以非水溶性成分对细胞活力影响更为明显。 2.PM2.5染毒可促进HUVEC细胞ACE、 AT1R基因表达,以非水溶性成分更为明显。培哚普利可抑制PM2.5染毒后ACE基因表达上调。
[Abstract]:Aim: to investigate the effects of PM2.5 on the activity of HUVECs and the expression of ACE, ATlR mRNA and protein in human umbilical vein endothelial cells (HUVECs), and to investigate the regulation of perindopril on the expression of ACE-AT1R in HUVEC cells. Methods: PM2.5 was collected from non-industrial area of Changsha from October 2010 to December 2011, and water soluble and insoluble components of PM2.5 were extracted. HUVEC cells were inoculated with 100 渭 g / ml of 100 渭 g / ml 200 渭 g / ml of 200 渭 g 路ml ~ (-1) or 400 渭 g / ml of 100 渭 g / ml (called low concentration, medium concentration, high concentration of water soluble or insoluble components of PM2.5) for 24 h. MTT assay was used to compare the effects of water soluble and insoluble components of PM2.5 on the viability of HUVEC cells. The expression of ACE-ATlR mRNA and protein in HUVEC cells was analyzed by real-time fluorescent quantitative PCR and Western Blot. HUVEC cells were exposed to high concentration PM2.5 for 24 h and 48 h respectively. The effects of PM2.5 on cell viability and ACE-AT1R gene expression were compared. At the same time, 10 渭 mol / L perindopril was added to HUVEC cells exposed to PM2.5 to investigate the effects of perindopril on the expression of ACE-AT1R in HUVEC cells. Results: 1. The inhibitory effect of PM2.5 on HUVEC cell viability: the survival rate of HUVEC cells in the water-soluble and insoluble components groups was lower than that in the blank control group (P 0.05), and that in the middle and low concentration groups in the high concentration group was lower than that in the low concentration group. At the same dose and high concentration, the effect of insoluble components on cell viability was significantly higher than that of water-soluble components. The cell survival rate of high concentration PM2.5 at different time groups was lower than that of 0 h group (P 0.05), and 48 h group 24 h group 12 h group (P 0.05) .2.The effect of PM2.5 on the expression of ACE gene in HUVEC cells: water soluble components. The expression of ACE mRNA and protein in the cells of high concentration and insoluble components groups were higher than that of control group (P 0.05), and the highest ACE expression level was in the group of high concentration of water soluble components (P 0.05). The expression of ACE gene was higher in high concentration water soluble components group (24 h or 48 h) than in Oh group (P 0.05), and the highest level of ACE expression was found in water soluble component group (24 h). 3. The effect of PM2.5 on AT1R gene expression in HUVEC cells. The expression of AT1R mRNA and protein in the high concentration of water-soluble and insoluble components was higher than that in the blank control group, and the highest expression of AT1R was found in the high concentration of water-soluble and insoluble components. The expression of AT1R in high concentration water soluble and insoluble components groups was higher than that in Oh group at different time points, and the highest AT1R expression level was found in 48 h insoluble components group. The effect of perindopril on the expression of ACE-AT1R in HUVEC cells: after perindopril treatment, the water-soluble and insoluble components were low, and the expression of ACEmRNA and protein in the medium and high concentration groups were significantly lower than those in the control group (P 0.05). However, the expression of AT 1 R mRNA and protein did not change significantly (P 0.05). Conclusion: 1.The activity of HUVEC cells was decreased by PM2.5 exposure, especially by the insoluble components. 2. PM2.5 could promote the expression of ACE-AT1R gene in HUVEC cells, especially the insoluble components. Perindopril inhibited the up-regulation of ACE gene expression after PM2.5 exposure.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

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