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  本文選題：Toll樣受體7 + Toll樣受體9��； 參考：《廣西醫(yī)科大學》2014年碩士論文

【摘要】：目的觀察TOLL樣受體7(Toll Like Receptor7, TLR7)、TOLL樣受體9(Toll Like Receptor9, TLR9)、核因子-κB P65（Nuclear Factor-κB P65, NF-κBP65）、腫瘤壞死因子-α（Tumor Necrosis Factor-α，TNF-α）在脂多糖誘導的AR42J細胞急性胰腺炎模型中的表達變化，探討TLR7、TLR9在急性胰腺炎發(fā)病機制中的作用及可能的作用機制。 方法取對數(shù)生長期的AR42J細胞用6孔細胞培養(yǎng)板傳代培養(yǎng)24小時后，分別用含脂多糖濃度為0mg/L、1mg/L、10mg/L、100mg/L的培養(yǎng)液刺激AR42J細胞，構建體外急性胰腺炎細胞模型,孵育24小時后，收集細胞培養(yǎng)上清液，離心留取上清，用ELISA法檢測細胞培養(yǎng)上清液中TNF-α的含量；收集細胞，提取細胞總RNA，用RT-PCR法分別檢測細胞中TLR7、TLR9、NF-κB P65mRNA的表達量變化；提取細胞總蛋白，用Western Blot法分別檢測細胞中TLR7、TLR9、NF-κB P65蛋白質(zhì)表達水平變化。 結果與脂多糖濃度為0mg/L的空白對照組相比，其余各組TLR7、TLR9、NF-κB P65的mRNA和蛋白表達量都隨著脂多糖濃度升高而增加，脂多糖濃度為100mg/L時達到最大值，各組間的差異有統(tǒng)計學意義（均P＜0.05）。TLR7、TLR9與NF-κB P65在mRNA、蛋白的表達水平上皆有正相關性（mRNA：r=0.786，0.762；蛋白：r=0.801，，0.720；均P＜0.05）。細胞培養(yǎng)上清液中TNF-α的含量呈脂多糖濃度依賴性增加，當脂多糖濃度為100mg/L時達到最大值，各組間的差異有統(tǒng)計學意義（P＜0.05）。 結論（1）脂多糖誘導的AR42J細胞急性胰腺炎模型中，TLR7及TLR9表達明顯上調(diào)，提示兩者可能在急性胰腺炎疾病發(fā)展中發(fā)揮重要作用。（2）TLR7、TLR9可能通過作用于NF-κB上調(diào)下游炎癥因子的產(chǎn)生，而參與急性胰腺炎的炎癥反應。
[Abstract]:Objective to observe the expression of TOLL like receptor 7(Toll Like Receptor7, TLR7TLL-like receptor 9(Toll Like Receptor9, nuclear factor- 魏 B P65(Nuclear Factor- 魏 B P65, NF- 魏 BP65, tumor necrosis factor- 偽 tumor Necrosis factor- 偽 in the model of AR42J cell acute pancreatitis induced by lipopolysaccharide (LPS). To investigate the role and possible mechanism of TLR7 and TLR9 in the pathogenesis of acute pancreatitis. Methods AR42J cells in logarithmic growth phase were cultured on a 6-well cell culture plate for 24 hours. The AR42J cells were stimulated with a culture medium containing a concentration of 0 mg / L of lipopolysaccharide at a concentration of 10 mg / L ~ (10) mg 路L ~ (-1). The acute pancreatitis cell model was established in vitro, and incubated for 24 hours. We collected the supernatant of cell culture, centrifuged the supernatant, detected the content of TNF- 偽 in the supernatant of cell culture by ELISA method, collected the cells, extracted the total RNAs of the cells, detected the changes of the expression of NF- 魏 B P65mRNA in the cells by RT-PCR method, and extracted the total cell protein. The expression level of NF- 魏 B p65 in TLR7 and TLR9 was detected by Western Blot assay. Results compared with the control group with lipopolysaccharide concentration of 0mg/L, the mRNA and protein expression of NF- 魏 B p65 in other groups increased with the increase of lipopolysaccharide concentration, and reached the maximum when the concentration of LPS was 100mg/L. There were significant differences among the three groups (P < 0.05). TLR7, TLR9 and NF- 魏 B p65 were all positively correlated with mRNAs in mRNAs, the protein expression levels were all positively correlated with mRNA-0.7860.762protein: rr-0.801and 0.720; all of them were all P < 0.05. The content of TNF- 偽 in the supernatant of cell culture was increased in a dose-dependent manner, and reached the maximum when the concentration of lipopolysaccharide was 100mg/L (P < 0.05). Conclusion the expression of TLR7 and TLR9 in acute pancreatitis model of AR42J cells induced by lipopolysaccharide is obviously up-regulated, which suggests that the expression of TLR7 and TLR9 may play an important role in the development of acute pancreatitis, which may be related to the up-regulation of down-stream inflammatory factor production by NF- 魏 B. And participate in the inflammatory response of acute pancreatitis.
【學位授予單位】：廣西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R57

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