Notch1在肌成纤维细胞转化中的作用及表达变化

发布时间：2018-02-09 11:51

本文关键词： Notch α-平滑肌肌动蛋白肌成纤维细胞肺纤维化　出处：《中国现代医学杂志》2016年15期 　论文类型：期刊论文

【摘要】：目的探讨在成纤维细胞转化为肌成纤维细胞过程中,Notch1表达的变化规律,以及γ-分泌酶抑制剂(DAPT)抑制Notch信号后,对细胞转化的影响。方法取新出生2~3 d SD大鼠的肺组织,用胰酶消化法分离肺成纤维细胞,再将培养至第3代的细胞分为3组:对照组、转化生长因子-β_1(TGF-β_1)组、TGF-β_1+DAPT组。对照组为空白对照,TGF-β_1组加入5ng/ml TGF-β_1,TGF-β_1+DAPT组同时加入5 ng/ml TGF-β_1及5μmol/L DAPT。采用免疫细胞化学法对α-平滑肌肌动蛋白(α-SMA)的表达变化进行检测。同时通过逆转录聚合酶链反应(RT-PCR)及Western blot检测Notch1 m RNA和蛋白的表达变化。结果α-SMA免疫细胞化学结果表明,对照组大部分细胞无染色,而TGF-β_1组则可见大部分细胞内有黄色和棕黄色颗粒及条纹,DAPT组和对照组无明显差异。对照组、TGF-β_1组、TGF-β_1+DAPT组Notch1 m RNA表达量分别为(0.278±0.022)、(0.783±0.018)和(0.313±0.029),对照组与TGF-β_1组、TGF-β_1组与TGF-β_1+DAPT组比较,差异有统计学意义(P0.05),对照组与TGF-β_1+DAPT组比较差异无统计学意义(P0.05)。对照组、TGF-β_1组、TGF-β_1+DAPT组Notch1蛋白表达量分别为(0.312±0.019)、(0.701±0.026)和(0.345±0.022),组间比较结果同Notch1 m RNA。结论 Notch1可以促进成纤维细胞向肌成纤维细胞的转化,进而促进肺纤维化。
[Abstract]:Objective to investigate the changes of Notch1 expression in fibroblasts and the effect of 纬 -secretase inhibitor (DAPT) on cell transformation after inhibiting Notch signal. Lung fibroblasts were isolated by trypsin digestion. The cells cultured to the third passage were divided into three groups: control group. TGF- 尾 1 DAPT group was added to TGF- 尾 1 DAPT group. The control group was added 5 ng / ml TGF- 尾 1 TGF- 尾 1 DAPT group to add 5 ng / ml TGF- 尾 1 DAPT group. The expression of 伪 -smooth muscle actin (伪 -SMA) was detected by immunocytochemistry. The expression of Notch1 m RNA and protein was detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot. Results 伪 -SMA immunocytochemistry showed that, Most of the cells in the control group were not stained. The expression of Notch1 m RNA in the TGF- 尾 1 DAPT group was 0.278 卤0.022 卤0.018 and 0.313 卤0.029 respectively. The comparison between the control group and the TGF- 尾 1 group showed that the expression of Notch1 m RNA in the control group and the TGF- 尾 1 group was higher than that in the TGF- 尾 1 group, and the expression of Notch1 m RNA in the TGF- 尾 1 DAPT group was higher than that in the TGF- 尾 1 DAPT group, and the expression of Notch1 m RNA in the TGF- 尾 1 DAPT group was higher than that in the TGF- 尾 1 DAPT group, while the expression of Notch1 m RNA in the TGF- 尾 1 DAPT group was 0.38 卤0.083 卤0.018 and 0.313 卤0.029 respectively. There was no significant difference between control group and TGF- 尾 1 DAPT group (P 0.05). The expression of Notch1 protein in TGF- 尾 1 DAPT group was 0.312 卤0.019 0. 701 卤0. 026 and 0. 345 卤0. 022 2 respectively. Conclusion Notch1 can promote fibroblasts. The transformation to myofibroblasts, Then promote pulmonary fibrosis.
【作者单位】：浙江省杭州市萧山区第一人民医院呼吸科;南昌大学第二附属医院呼吸科;
【基金】：江西省自然科学基金(No:20132BAB205014)
【分类号】：R563

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