本文選題：脂多糖　切入點：血管內皮細胞　出處：《安徽醫(yī)科大學》2017年碩士論文　論文類型：學位論文

【摘要】：目的探討Tribbles同源蛋白3(TRB3)在脂多糖(LPS)致急性肺損傷時的變化及其與p38-MAPK信號通路的關系。方法體內實驗:復制LPS致急性肺損傷(ALI)大鼠模型,分生理鹽水對照組(5ml/kg)和5 mg/kg LPS刺激組,免疫組織化學法檢測肺組織中TRB3蛋白表達,逆轉錄聚合酶聯(lián)反應(RT-PCR)檢測肺組織TRB3 m RNA表達。體外實驗:體外培養(yǎng)大鼠肺微血管內皮細胞(PMVEC),隨機分為LPS量效組(0、2、4、10μg/ml LPS分別孵育4 h)、LPS時效組(10μg/ml LPS分別孵育0、4、8、12 h)和p38抑制劑(SB203580)干預組:分為正常對照組,10μg/ml LPS組、10μmol/L SB203580組、10μmol/L SB203580+10μg/ml LPS組,免疫印跡法檢測TRB3蛋白、p-p38和p38-MAPK表達。結果免疫組織化學法顯示大鼠肺泡壁和腺上皮均表達TRB3;RT-PCR法檢測大鼠肺組織和大鼠PMVEC均表達TRB3 m RNA;與生理鹽水組比較,LPS致ALI大鼠肺組織TRB3 m RNA表達顯著增加(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表達增加(2.098±0.317 vs0.612±0.314,t=7.549,P0.01);免疫印跡法發(fā)現PMVEC表達TRB3蛋白:表達量隨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)計學意義(F=12.619,P0.001);時效組TRB3蛋白表達量于4 h達高峰(0.443±0.087),之后下降, 8 h(0.303±0.107),仍高于正常對照組(0.159±0.073),組間比較,差異有統(tǒng)計學意義(F=11.273,P0.001)。干預組:與正常對照組比較,10μg/ml LPS誘導大鼠PMVEC的p-p38蛋白表達量增高(0.660±0.100 vs 0.227±0.085,t=49.121,P0.001)以及TRB3蛋白表達增高(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單獨刺激組比較,p-p38蛋白表達量下降(0.557±0.125 vs 0.660±0.100,t=7.040,P0.05,TRB3表達量也下降(0.306±0.077 vs 0.461±0.097,t=11.900,P0.001);SB203580單獨作用對大鼠PMVEC表達p-p38及TRB3蛋白無影響(與正常對照組比較,P0.05)。結論LPS致急性肺損傷時TRB3表達增加,TRB3表達受p38-MAPK信號通路調控。
[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.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R563.8

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相關期刊論文 前1條

1 張丹;尤青海;孫耕耘;王楠;岳揚;邵敏;;脂多糖誘導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;2013年01期

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本文編號：1580459