本文選題：TGF-β1 + 支氣管上皮細(xì)胞　； 參考：《青島大學(xué)》2017年碩士論文

【摘要】：目的:探討TGF-β1對人支氣管上皮細(xì)胞間充質(zhì)轉(zhuǎn)化及Six1基因表達的影響。方法:體外培養(yǎng)人支氣管上皮細(xì)胞(16HBE),構(gòu)建上皮-間充質(zhì)轉(zhuǎn)化模型。采用倒置顯微鏡觀察各組細(xì)胞形態(tài)的變化;采用Real-time PCR檢測E-鈣黏蛋白、波形蛋白、α-肌動蛋白、纖維黏連蛋白及Six1基因m RNA表達的情況;采用Westernblotting檢測E-鈣黏素、波形蛋白、α-肌動蛋白、纖維黏連蛋白及Six1蛋白的表達變化。結(jié)果:1.倒置顯微鏡結(jié)果示:對照組細(xì)胞呈單層,細(xì)胞間連接較為緊密;處理組A細(xì)胞形態(tài)變化不明顯;處理組B部分細(xì)胞出現(xiàn)肥大、拉長,細(xì)胞間連接緊密性下降;處理組C細(xì)胞形態(tài)變化較為明顯,呈梭行,同時細(xì)胞間隙增大,喪失典型的鋪路石樣特征。2.RT-PCR檢測結(jié)果示:對照組細(xì)胞可正常表達E-鈣黏素基因、波形蛋白基因、纖維黏連蛋白基因、α-肌動蛋白基因及Six1基因m RNA;與對照組相比較,處理組A其E-鈣粘素基因、α-肌動蛋白基因m RNA表達量降低(P值分別為0.862、0.566,均0.05),Six1基因m RNA表達量增高(P值為0.1020.05),差異均無統(tǒng)計學(xué)意義,波形蛋白基因、纖維黏連蛋白基因m RNA表達量顯著升高(P值分別為0.003、0.0060.01),差異有統(tǒng)計學(xué)意義。處理組B其E-鈣粘素基因m RNA表達量明顯減少(P=0.042),波形蛋白基因、纖維黏連蛋白基因、α-肌動蛋白基因及Six1基因m RNA表達量均顯著增加(P=0.000、0.001、0.031、0.023,均小于0.05),與對照組相比差異均有統(tǒng)計學(xué)意義。處理組C其E-鈣粘素基因m RNA表達量顯著減少(P=0.0040.01),波形蛋白基因、纖維黏連蛋白基因、α-肌動蛋白基因及Six1基因m RNA表達量均明顯增加(P=0.004、0.003、0.023、0.004,均小于0.05),與對照組相比差異均有統(tǒng)計學(xué)意義。3.Westernblot檢測結(jié)果示:對照組細(xì)胞能夠正常表達E-鈣粘素、α-肌動蛋白、纖維黏連蛋白、波形蛋白及Six1蛋白。與對照組相比,處理組A細(xì)胞中E-鈣粘素表達減少(P=1.0230.05),α-肌動蛋白、纖維黏連蛋白及波形蛋白的表達量增高(P值分別為0.789、0.542、0.624,均0.05),但差異均無統(tǒng)計學(xué)意義,Six1蛋白表達量增加(P=0.0280.05),差異有統(tǒng)計學(xué)意義;處理組B細(xì)胞中E-鈣粘素表達量明顯減少(P=0.0110.05),α-肌動蛋白、纖維黏連蛋白、波形蛋白及Six1蛋白的表達量均顯著增加(P值分別為0.002、0.001、0.004、0.009均0.01),與對照組相比差異均有統(tǒng)計學(xué)意義;處理組C細(xì)胞中E-鈣粘素表達量顯著減少(P=0.0090.01),α-肌動蛋白、波形蛋白及Six1蛋白的表達量顯著增加(P值分別為0.008、0.003、0.007,均0.01),纖維黏連蛋白表達量明顯增加(P值為0.0170.05),與對照組相比差異均有統(tǒng)計學(xué)意義。且隨TGF-β1作用時間越長,Six1蛋白表達越高,各組間有顯著性差異(F=33.50)。結(jié)論:TGF-β1可能通過上調(diào)Six1的表達誘導(dǎo)氣道上皮細(xì)胞間充質(zhì)轉(zhuǎn)化,從而參與哮喘氣道重塑。
[Abstract]:Aim: to investigate the effect of TGF- 尾 1 on mesenchymal transformation and expression of Six1 gene in human bronchial epithelial cells. Methods: human bronchial epithelial cells were cultured in vitro to construct epithelial-mesenchymal transformation model. The morphological changes of each group were observed by inverted microscope, the expressions of Ecadherin, vimentin, 偽 -actin, fibronectin and Six1 mRNA were detected by Real-time PCR, and Ecadherin was detected by Western blotting. The expression of vimentin, 偽-actin, fibronectin and Six1 protein. The result is 1: 1. The results of inverted microscope showed that the cells of the control group were monolayer and the intercellular junctions were close; the morphological changes of the A cells in the treatment group were not obvious; the cells in part B of the treatment group were hypertrophic, elongated, and the tight intercellular junctions decreased. The morphological changes of C cells in the treatment group were obvious, and the cell gap was enlarged, and the typical paving stone features were lost. 2. The results of RT-PCR showed that the normal expression of Ecadherin gene and vimentin gene were observed in the control group. Fibronectin gene, 偽 -actin gene and Six1 gene mRNAs were compared with control group. In the treatment group A, the mRNA expression of E-cadherin gene and 偽 -actin gene decreased by 0.862 ~ 0.566.The mRNA expression of each group was increased (P = 0.1020.05, P = 0.1020.05, respectively), vimentin gene. The mRNA expression of fibronectin gene was significantly increased (P = 0.003, P = 0.0060.01), and the difference was statistically significant. In treatment group B, the expression of Ecadherin gene mRNA was significantly reduced by P0.042, vimentin gene. The mRNA expression of fibronectin gene, 偽 -actin gene and Six1 gene were all increased significantly (P < 0.05). In treatment group C, the expression of Ecadherin gene mRNA was significantly reduced by P0. 0040.01, vimentin gene. The mRNA expression of fibronectin gene, 偽 -actin gene and Six1 gene were all increased significantly (P < 0.05). Compared with the control group, the expression of E-cadherin and 偽 -actin in the control group was significantly higher than that in the control group. The results of Western blot showed that the normal expression of E-cadherin and 偽 -actin could be observed in the control group, and the expression of E-cadherin and 偽 -actin in the control group was significantly higher than that in the control group. Fibronectin, vimentin and Six1 protein. Compared with the control group, the expression of E-cadherin in A cells of the treatment group was decreased by P1.0230.05, 偽 -actin. The expression of fibronectin and vimentin were increased (P = 0.789) 0.542n 0.624, respectively (0.05%), but there was no significant difference in the expression of Six1 protein (P < 0.05), the difference was statistically significant, and the expression of E-cadherin in B cells decreased significantly (P 0.0110.05, 偽 -actin, 偽 -actin, P < 0.05), the expression of E-cadherin and vimentin in B cells was significantly lower than that in control group (P < 0.05), but there was no significant difference in the expression of Six1 protein (P < 0.05). The expression of fibronectin, vimentin and Six1 protein increased significantly (P = 0.002, 0.001, 0.004, 0.009, respectively), which were significantly different from those of the control group, and the expression of Ecadherin in C cells of treatment group decreased significantly, and the expression of Ecadherin and 偽 -actin in C cells of treatment group were significantly lower than that of control group, and the expression of Ecadherin in C cells of treatment group was significantly lower than that of control group (P < 0.05). The expression of vimentin and Six1 protein increased significantly (P = 0.008, P = 0.003, P = 0.01g, P = 0.0170.05, P = 0.0170.05, respectively). The higher the expression of TGF- 尾 1 protein was, the higher the expression of TGF- 尾 1 protein was. Conclusion: TGF- 尾 1 may be involved in airway remodeling by up-regulating the expression of Six1 and inducing airway epithelial mesenchymal transformation.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R725.6

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