本文選題：脂多糖　切入點(diǎn)：血管內(nèi)皮細(xì)胞　出處：《安徽醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的探討Tribbles同源蛋白3(TRB3)在脂多糖(LPS)致急性肺損傷時(shí)的變化及其與p38-MAPK信號(hào)通路的關(guān)系。方法體內(nèi)實(shí)驗(yàn):復(fù)制LPS致急性肺損傷(ALI)大鼠模型,分生理鹽水對(duì)照組(5ml/kg)和5 mg/kg LPS刺激組,免疫組織化學(xué)法檢測(cè)肺組織中TRB3蛋白表達(dá),逆轉(zhuǎn)錄聚合酶聯(lián)反應(yīng)(RT-PCR)檢測(cè)肺組織TRB3 m RNA表達(dá)。體外實(shí)驗(yàn):體外培養(yǎng)大鼠肺微血管內(nèi)皮細(xì)胞(PMVEC),隨機(jī)分為LPS量效組(0、2、4、10μg/ml LPS分別孵育4 h)、LPS時(shí)效組(10μg/ml LPS分別孵育0、4、8、12 h)和p38抑制劑(SB203580)干預(yù)組:分為正常對(duì)照組,10μg/ml LPS組、10μmol/L SB203580組、10μmol/L SB203580+10μg/ml LPS組,免疫印跡法檢測(cè)TRB3蛋白、p-p38和p38-MAPK表達(dá)。結(jié)果免疫組織化學(xué)法顯示大鼠肺泡壁和腺上皮均表達(dá)TRB3;RT-PCR法檢測(cè)大鼠肺組織和大鼠PMVEC均表達(dá)TRB3 m RNA;與生理鹽水組比較,LPS致ALI大鼠肺組織TRB3 m RNA表達(dá)顯著增加(3.675±0.423 vs 1.056±0.209,t=15.524,P0.01);與未刺激組比較,LPS刺激的大鼠PMVEC中TRB3 m RNA表達(dá)增加(2.098±0.317 vs0.612±0.314,t=7.549,P0.01);免疫印跡法發(fā)現(xiàn)PMVEC表達(dá)TRB3蛋白:表達(dá)量隨LPS濃度(0、2、4、10μg/ml)增加逐漸升高,分別為0.169±0.089、0.198±0.071、0.338±0.089、0.494±0.118,組間比較,差異有統(tǒng)計(jì)學(xué)意義(F=12.619,P0.001);時(shí)效組TRB3蛋白表達(dá)量于4 h達(dá)高峰(0.443±0.087),之后下降, 8 h(0.303±0.107),仍高于正常對(duì)照組(0.159±0.073),組間比較,差異有統(tǒng)計(jì)學(xué)意義(F=11.273,P0.001)。干預(yù)組:與正常對(duì)照組比較,10μg/ml LPS誘導(dǎo)大鼠PMVEC的p-p38蛋白表達(dá)量增高(0.660±0.100 vs 0.227±0.085,t=49.121,P0.001)以及TRB3蛋白表達(dá)增高(0.461±0.097 vs 0.178±0.084,t=15.113,P0.001);10μmol/L SB203580+10μg/ml LPS聯(lián)合刺激組與LPS單獨(dú)刺激組比較,p-p38蛋白表達(dá)量下降(0.557±0.125 vs 0.660±0.100,t=7.040,P0.05,TRB3表達(dá)量也下降(0.306±0.077 vs 0.461±0.097,t=11.900,P0.001);SB203580單獨(dú)作用對(duì)大鼠PMVEC表達(dá)p-p38及TRB3蛋白無影響(與正常對(duì)照組比較,P0.05)。結(jié)論LPS致急性肺損傷時(shí)TRB3表達(dá)增加,TRB3表達(dá)受p38-MAPK信號(hào)通路調(diào)控。
[Abstract]:Objective to investigate the changes of Tribbles homologue protein 3 (TRB3) in acute lung injury induced by lipopolysaccharide (LPS) and its relationship with p38-MAPK signaling pathway. Methods the rat model of acute lung injury induced by LPS was established in vivo and divided into normal saline control group (5 ml / kg) and 5 mg/kg LPS stimulation group. Immunohistochemical method was used to detect the expression of TRB3 protein in lung tissue. Reverse transcriptase polymerase chain reaction (RT PCR) was used to detect the expression of TRB3 m RNA in lung tissue. In vitro, rat pulmonary microvascular endothelial cells were incubated with 10 渭 g / ml LPS (10 渭 g / ml LPS) and p38 (10 渭 g / ml LPS, respectively) and p38. The control group was divided into 10 渭 g / ml LPS group and 10 渭 mol/L SB203580 10 渭 g / ml LPS group, 10 渭 mol/L SB203580 group, 10 渭 mol/L SB203580 10 渭 g / ml LPS group, 10 渭 mol/L SB203580 10 渭 g / ml LPS group, 10 渭 mol/L SB203580 10 渭 g / ml LPS group. The expression of p-p38 and p38-MAPK in rat alveolar wall and glandular epithelium was detected by immunoblotting assay. Results the expression of TRB3 mRNA in rat lung tissue and rat PMVEC was detected by RT-PCR, and compared with that in normal saline group, the expression of TRB3 mRNA in rat alveolar wall and glandular epithelium was detected by immunohistochemistry. The expression of TRB3 m RNA increased significantly in lung tissue of rats (3.675 鹵0.423 vs 1.056 鹵0.209), and the expression of TRB3 m RNA in PMVEC was increased by 2.098 鹵0.317 vs0.612 鹵0.314 vs0.612 鹵7.549P0.01.The expression of TRB3 protein was increased with the increase of LPS concentration (10 渭 g / ml). The expression of TRB3 protein was 0.169 鹵0.089 鹵0.198 鹵0.071, 0.338 鹵0.089 and 0.494 鹵0.118, respectively. The difference between the two groups was statistically significant, and the expression of TRB3 protein reached the peak at 4 h, reached the peak at 4 h, then decreased, and decreased to 0.303 鹵0.107 at 8 h, which was still higher than that of the normal control group (0.159 鹵0.073). In the intervention group, the expression of p-p38 protein in PMVEC induced by 10 渭 g / ml LPS was significantly higher than that in the control group (0.660 鹵0.100 vs 0.227 鹵0.085) and the expression of TRB3 protein was 0.461 鹵0.097 vs 0.178 鹵0.084 渭 mol/L SB203580 10 渭 g / ml LPS combined with LPS alone. The decrease of protein expression (0.557 鹵0.125 vs 0.660 鹵0.100) P0.05TRB3 also decreased 0.306 鹵0.077 vs 0.461 鹵0.097 T11.900P0.001P0.001TRB3. The expression of p-p38 and TRB3 protein in PMVEC was not affected by LPS alone. Conclusion the increased expression of TRB3 in acute lung injury induced by LPS is regulated by p38-MAPK signaling pathway.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R563.8

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 張丹;尤青海;孫耕耘;王楠;岳揚(yáng);邵敏;;脂多糖誘導(dǎo)急性肺損傷大鼠肺組織小窩蛋白-1的變化[J];中國急救醫(yī)學(xué);2013年01期

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本文編號(hào)：1580459