本文關鍵詞： Notch α-平滑肌肌動蛋白 肌成纖維細胞 肺纖維化　出處：《中國現(xiàn)代醫(yī)學雜志》2016年15期 　論文類型：期刊論文

【摘要】：目的探討在成纖維細胞轉(zhuǎn)化為肌成纖維細胞過程中,Notch1表達的變化規(guī)律,以及γ-分泌酶抑制劑(DAPT)抑制Notch信號后,對細胞轉(zhuǎn)化的影響。方法取新出生2~3 d SD大鼠的肺組織,用胰酶消化法分離肺成纖維細胞,再將培養(yǎng)至第3代的細胞分為3組:對照組、轉(zhuǎn)化生長因子-β_1(TGF-β_1)組、TGF-β_1+DAPT組。對照組為空白對照,TGF-β_1組加入5ng/ml TGF-β_1,TGF-β_1+DAPT組同時加入5 ng/ml TGF-β_1及5μmol/L DAPT。采用免疫細胞化學法對α-平滑肌肌動蛋白(α-SMA)的表達變化進行檢測。同時通過逆轉(zhuǎn)錄聚合酶鏈反應(RT-PCR)及Western blot檢測Notch1 m RNA和蛋白的表達變化。結(jié)果α-SMA免疫細胞化學結(jié)果表明,對照組大部分細胞無染色,而TGF-β_1組則可見大部分細胞內(nèi)有黃色和棕黃色顆粒及條紋,DAPT組和對照組無明顯差異。對照組、TGF-β_1組、TGF-β_1+DAPT組Notch1 m RNA表達量分別為(0.278±0.022)、(0.783±0.018)和(0.313±0.029),對照組與TGF-β_1組、TGF-β_1組與TGF-β_1+DAPT組比較,差異有統(tǒng)計學意義(P0.05),對照組與TGF-β_1+DAPT組比較差異無統(tǒng)計學意義(P0.05)。對照組、TGF-β_1組、TGF-β_1+DAPT組Notch1蛋白表達量分別為(0.312±0.019)、(0.701±0.026)和(0.345±0.022),組間比較結(jié)果同Notch1 m RNA。結(jié)論 Notch1可以促進成纖維細胞向肌成纖維細胞的轉(zhuǎn)化,進而促進肺纖維化。
[Abstract]:Objective to investigate the changes of Notch1 expression in fibroblasts and the effect of 緯 -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 偽 -smooth muscle actin (偽 -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 偽 -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 鹵0.022 鹵0.018 and 0.313 鹵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 鹵0.083 鹵0.018 and 0.313 鹵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 鹵0.019 0. 701 鹵0. 026 and 0. 345 鹵0. 022 2 respectively. Conclusion Notch1 can promote fibroblasts. The transformation to myofibroblasts, Then promote pulmonary fibrosis.
【作者單位】： 浙江省杭州市蕭山區(qū)第一人民醫(yī)院呼吸科;南昌大學第二附屬醫(yī)院呼吸科;
【基金】：江西省自然科學基金(No:20132BAB205014)
【分類號】：R563

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