本文選題：細(xì)顆粒物 + 血管內(nèi)皮細(xì)胞　； 參考：《中南大學(xué)》2012年碩士論文

【摘要】：目的： 分析細(xì)顆粒物(PM2.5)對人臍靜脈內(nèi)皮細(xì)胞(HUVEC)活力和ACE、 ATlR mRNA和蛋白表達(dá)的影響,探討培哚普利對HUVEC細(xì)胞ACE、 AT1R表達(dá)的調(diào)控。 方法： 于2010年10月-2011年12月采集長沙市非工業(yè)區(qū)大氣的PM2.5,并提取PM2.5的水溶性和非水溶性成分。分別用100μg/ml、200μg/ml、400μg/ml(稱為低濃度、中濃度、高濃度)PM2.5水溶性和非水溶性成分染毒HUVEC細(xì)胞24h。采用MTT法比較不同濃度PM2.5水溶性和非水溶性成分染毒對HUVEC細(xì)胞活力的影響,采用實時熒光定量PCR和Western Blot方法分析HUVEC細(xì)胞ACE、 ATlR mRNA和蛋白表達(dá)的差異。利用高濃度PM2.5染毒HUVEC細(xì)胞Oh、12h、24h和48h,比較不同時間PM2.5染毒對細(xì)胞活力和ACE, AT1R基因表達(dá)的影響。同時在PM2.5染毒HUVEC細(xì)胞時加入10μmol/L的培哚普利進(jìn)行干預(yù),分析培哚普利對HUVEC細(xì)胞ACE, AT1R表達(dá)的調(diào)控。 結(jié)果： 1.PM2.5對HUVEC細(xì)胞活力的抑制效應(yīng)：水溶性和非水溶性成分各濃度組細(xì)胞存活率均低于空白對照組(P0.05),且高濃度組中濃度組低濃度組(P0.05)。當(dāng)染毒劑量同為中、高濃度時,非水溶性成分對細(xì)胞活力影響較水溶性成分明顯(P0.05)。高濃度PM2.5染毒不同時間組細(xì)胞存活率均低于0h組(P0.05),且48h組24h組12h組(P0.05)。 2.PM2.5對HUVEC細(xì)胞ACE基因表達(dá)的影響：水溶性成分中、高濃度組和非水溶性成分各濃度組細(xì)胞ACEmRNA和蛋白表達(dá)量均高于空白對照組(P0.05),且非水溶性成分高濃度組ACE表達(dá)量最高(P0.05)。高濃度水溶性成分24h、48h組和非水溶性成分各不同時間組細(xì)胞ACE基因表達(dá)量高于Oh組(P0.05),且非水溶性成分24h組ACE表達(dá)量最高(P0.05)。 3.PM2.5對HUVEC細(xì)胞AT1R基因表達(dá)的影響：水溶性和非水溶性成分高濃度組細(xì)胞AT1RmRNA和蛋白表達(dá)量高于空白對照組(P0.05),且非水溶性成分高濃度組AT1R表達(dá)量最高(P0.05)。高濃度水溶性和非水溶性成分各不同時間組細(xì)胞AT1R表達(dá)量高于Oh組(P0.05),且非水溶性成分48h組AT1R表達(dá)量最高(P0.05)。 4.培哚普利干預(yù)對HUVEC細(xì)胞ACE、 AT1R表達(dá)的調(diào)控：培哚普利干預(yù)后,水溶性和非水溶性成分低、中、高濃度組細(xì)胞ACEmRNA和蛋白表達(dá)量均顯著低于未加藥物組(P0.05)；但AT1R mRNA和蛋白表達(dá)量無顯著改變(P0.05)。 結(jié)論： 1.PM2.5染毒可導(dǎo)致HUVEC細(xì)胞活力降低,以非水溶性成分對細(xì)胞活力影響更為明顯。 2.PM2.5染毒可促進(jìn)HUVEC細(xì)胞ACE、 AT1R基因表達(dá),以非水溶性成分更為明顯。培哚普利可抑制PM2.5染毒后ACE基因表達(dá)上調(diào)。
[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.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R363

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