發(fā)布時(shí)間：2018-05-04 00:03

  本文選題：妊娠相關(guān)血清蛋白-A + 內(nèi)皮細(xì)胞��； 參考：《中南大學(xué)》2008年碩士論文

【摘要】： 目的觀察妊娠相關(guān)血清蛋白-A(PAPP-A)在體外對(duì)內(nèi)皮細(xì)胞功能的影響,同時(shí)探討氟伐他汀對(duì)PAPP-A誘導(dǎo)內(nèi)皮細(xì)胞功能改變的干預(yù)作用。 方法體外培養(yǎng)人臍靜脈內(nèi)皮細(xì)胞株(HUVEC),將對(duì)數(shù)生長期的HUVEC隨機(jī)分為四組。分別給予不同濃度的PAPP-A(0 ng/ml、50ng/ml、100 ng/ml、200 ng/ml)刺激,于0h、12 h、24 h和48 h收集上清液和細(xì)胞,用硝酸還原酶法測定一氧化氮(NO)的濃度,用免疫組化法測定內(nèi)皮素-1(ET-1)的表達(dá)。取對(duì)數(shù)生長期的HUVEC隨機(jī)分為五組,實(shí)驗(yàn)組分別予不同濃度的氟伐他汀(0 mmol/L、10~(-7)mmol/L、10~(-6)mmol/L、10~(-5)mmol/L)干預(yù)20min后,加入200 ng/mlPAPP-A,12h后收集上清液和細(xì)胞,用硝酸還原酶法測定NO的濃度,用免疫組化法測定ET-1的表達(dá)。 結(jié)果在50ng/mlPAPP-A、100ng/mlPAPP-A及200ng/mlPAPP-A干預(yù)組,12h、24h和48h上清液NO濃度均低于同組0h NO的濃度,差異具有統(tǒng)計(jì)學(xué)意義,P＜0.05。在12h、24h和48h,隨著PAPP-A濃度的增加,上清液NO的濃度逐漸降低,與對(duì)照組相比具有統(tǒng)計(jì)學(xué)意義,P＜0.05。在200ng/mlPAPP-A干預(yù)組,細(xì)胞ET-1的表達(dá)呈時(shí)間依賴性增加;在12h、24h和48h,隨著PAPP-A濃度的增加,ET-1的表達(dá)呈濃度依賴性增加,與對(duì)照組相比具有統(tǒng)計(jì)學(xué)意義,P＜0.05。 實(shí)驗(yàn)第二步,隨著氟伐他汀干預(yù)濃度增加,上清液NO的濃度逐漸升高,與單純PAPP-A干預(yù)組比較具有統(tǒng)計(jì)學(xué)意義,P＜0.05。在單純PAPP-A干預(yù)組,細(xì)胞ET-1的表達(dá)明顯增加,與對(duì)照組比較具有統(tǒng)計(jì)學(xué)差異,P＜0.05。隨著氟伐他汀干預(yù)濃度增加,ET-1的表達(dá)逐漸減低,均低于單純PAPP-A干預(yù)組,P＜0.05,但仍高于正常對(duì)照組,P＜0.05。 結(jié)論P(yáng)APP-A在體外能誘導(dǎo)內(nèi)皮細(xì)胞NO分泌減少,ET-1表達(dá)上調(diào),且具有濃度依賴性,無明顯時(shí)間依賴性。氟伐他汀對(duì)PAPP-A誘導(dǎo)的內(nèi)皮細(xì)胞功能損害可能有保護(hù)作用,且具有濃度依賴性。
[Abstract]:Objective to observe the effect of pregnancy-associated serum protein (APP-A) on the function of endothelial cells in vitro, and to explore the effect of fluvastatin on the changes of endothelial cell function induced by PAPP-A. Methods Human umbilical vein endothelial cells (HUVECs) were cultured in vitro. HUVEC in logarithmic phase was randomly divided into four groups. The supernatants and cells were collected at 0 h ~ 12 h for 24 h and 48 h respectively. The concentration of nitric oxide (no) was determined by nitrate reductase method. The expression of endothelin-1 ET-1 was detected by immunohistochemical method. HUVEC in logarithmic growth phase was randomly divided into five groups. The experimental group was treated with different concentrations of fluvastatin 0 mmol / L 10 ~ 7 mmol / L ~ (10) ~ (-1) ~ (-1) ~ (-1) ~ (-1) / L ~ (10) mol / L ~ (10) ~ (-1) 20min, then added 200ng / ml PAPP-A for 12 h to collect supernatant and cell, and then the concentration of no was determined by nitrate reductase method, and the concentration of no was determined by nitric acid reductase method, and the concentration of no was determined by nitrate reductase method. The expression of ET-1 was detected by immunohistochemistry. Results at 50 ng / ml PAPP-A 100 ng / ml PAPP-A and 200ng/mlPAPP-A intervention group, the concentration of no in supernatant of 12h and 48h was lower than that of the same group (P < 0.05). At 12h and 48h, the concentration of no in supernatant gradually decreased with the increase of PAPP-A concentration, which was statistically significant compared with the control group (P < 0.05). In the 200ng/mlPAPP-A intervention group, the expression of ET-1 increased in a time-dependent manner, and increased in a concentration-dependent manner with the increase of PAPP-A concentration at 12h and 48h, which was significantly higher than that in the control group (P < 0.05). In the second step of the experiment, with the increase of fluvastatin concentration, the concentration of no in supernatant gradually increased, which was statistically significant compared with that of PAPP-A group (P < 0.05). The expression of ET-1 in PAPP-A group was significantly higher than that in control group (P < 0.05). With the increase of fluvastatin concentration, the expression of Tet-1 decreased gradually, which was lower than that of PAPP-A group (P < 0. 05), but still higher than that of normal control group (P < 0. 05). Conclusion in vitro, PAPP-A can induce no secretion in endothelial cells to decrease the expression of Tet-1 in a dose-dependent manner, but not in time dependent manner. Fluvastatin may have a protective effect on PAPP-A induced endothelial cell dysfunction, and it is in a concentration dependent manner.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R363

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本文編號(hào)：1840668