TGF-β1 activates mouse hepatic stellate cells through Jagged1/Notch

Ai Maiti · Ya Sen; Jin Xin; Wang Wei; Li Dewei

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TGF-β1 activates mouse hepatic stellate cells through Jagged1/Notch

更新时间：2023-12-22

- TGF-β1 activates mouse hepatic stellate cells through Jagged1/Notch
- Journal of Chongqing Medical University Vol. 43, Issue 5, Pages: 634(2018)
- 作者机构：
  
  重庆医科大学附属第一医院肝胆外科、重庆医科大学第一附属医院实验研究中心,重庆,400016
- 作者简介：
- 基金信息：
- DOI：
  CLC：
- Published：2018，
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Ai Maiti · Ya Sen, Jin Xin, Wang Wei, et al. TGF-β1 activates mouse hepatic stellate cells through Jagged1/Notch. [J]. Journal of Chongqing Medical University 43(5):634(2018)
DOI：

Ai Maiti · Ya Sen, Jin Xin, Wang Wei, et al. TGF-β1 activates mouse hepatic stellate cells through Jagged1/Notch. [J]. Journal of Chongqing Medical University 43(5):634(2018) DOI：

摘要

目的：研究转化生长因子-β1（transforming growth factor β1，TGF-β1）激活小鼠肝星状细胞（mouse hepatic stellate cells，mHSC）的具体机制。方法：实验选用mHSC-T25细胞株为实验对象，随机分为激活组（mHSC-T25+10 ng/mL TGF-β1）、抑制组（mHSC-T25+1 μmol/L TGF-β-R1抑制剂）和对照组（mHSC-T25）。qRT-PCR检测α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）、TGF-β1、转化生长因子β受体1（transforming growth factorβreceptor1，TGF-β-R1）、Jagged1、血管内皮生长因子（vas－cular endothelial growth factor，VEGF）、肝细胞生长因子（hepatocyte growth factor，HGF）等的mRNA表达；细胞免疫荧光法检测α-SMA、Jagged1等蛋白的表达；Western blot检测TGF-β1、α-SMA、TGF-β-R1、Jagged1、Smad2/3、p-Smad2/3等蛋白的表达。结果：①TGF-β1刺激组中α-SMA、TGF-β1、TGF-β-R1、Jagged1、VEGF、HGF等mRNA表达量均较对照组明显增加而抑制组均明显下降。α-SMA：激活组（315.1±6.2）%，抑制组（34.3±4.5）% （F=3291.956，P=0.000）；TGF-β1：激活组（524.8±14.3）%，抑制组（29.0±10.7）%（F=469.534，P=0.000）；TGF-β-R1：激活组（235.5±15.2）%，抑制组（17.0±2.8）%（F=392.239，P=0.000）；Jagged1：激活组（548.7±16.6）%，抑制组（17.4±4.7）%（F=364.166，P=0.000）；VEGF：激活组（331.9±19.8）%，抑制组（19.5±3.7）%（F=278.407，P=0.000）；HGF：激活组（376.00±6.51）%，抑制组（16.2±2.7）%（F=640.340，P=0.000）。②激活组高表达α-SMA、Jagged1等蛋白，而对照组、抑制组表达明显较少。α-SMA：激活组0.880±0.016，对照组0.481±0.007，抑制组0.207±0.014 （F=2098.556，P=0.000）；Jagged1：激活组0.796±0.015，对照组0.474±0.021，抑制组0.167±0.005（F=1242.556，P=0.000）。③TGF-β1激活组中TGF-β1、α-SMA、TGF-β-R1、Jagged1、Smad2/3、p-Smad2/3的蛋白表达量均较对照组明显上调而抑制组均明显下调。TGF-β1：激活组1.180±0.138，对照组0.654±0.061，抑制组0.359±0.012（F=67.706，P=0.000）；α-SMA：激活组1.076±0.063，对照组0.689±0.022，抑制组0.374±0.011（F=239.186，P=0.000）；TGF-β-R1：激活组1.192±0.142，对照组0.710±0.380，抑制组0.378±0.069 （F=57.235，P=0.000）；Jagged1：激活组1.582±0.247，对照组0.817±0.116，抑制组0.364±0.059（F=43.649，P=0.000）；Smad2/3：激活组2.057±0.114，对照组1.203±0.105，抑制组0.664±0.063（F=158.856，P=0.000）；p-Smad2/3：激活组2.088±0.120，对照组1.168±0.107，抑制组0.573±0.120 （F=136.369，P=0.000）。上述实验结果提示，激活组、抑制组与对照组比较，差异均有统计学意义（P＜0.05）。结论：TGF-β1通过Jagged1/Notch激活小鼠肝星状细胞并调控其相关生物学活性。

Abstract

Objective：To study specific mechanisms of activation of mouse hepatic stellate cells（mHSCs） by transforming growth factor β1（TGF-β1）. Methods：mHSC-T25 cell line was selected as experimental subject，and randomly divided into activation group（mH－SC-T25+10 ng/mL TGF-β1），inhibition group（mHSC-T25+l μmol/L TGF-β-R1 inhibitor） and control group（mHSC-T25）. mRNA expression of α-SMA，TGF-β1，TGF-β-R1，Jagged1，VEGF，HGF were detected by qRT-PCR. Expressions of α-SMA，Jagged1 were measured by immunocytochemistory. Protein expressions of TGF-β1，α-SMA，TGF-β-R1，Jagged1，Smad2/3，p-Smad2/3 were tested by Western blot. Results：①mRNA expressions of α-SMA，TGF-β1，TGF-β-R1，Jagged1，VEGF，HGF were significantly higher in ac－tivation group than in control group，but significantly lower in inhibition group. Activation group：（315.1±6.2）%，（524.8±14.3）%，（235.5±15.2）%，（548.7±16.6）%，（331.9±19.8）%，（376.00±6.51）%；inhibition group：（34.3±4.5）%，（29.0±10.7）%，（17.0±2.8）%，（17.4±4.7）%，（19.5±3.7）%，（16.2±2.7）%（P=0.000）. ②Expressions of α-SMA，Jagged1 were significantly higher in activation group than in control group and inhibition group. Activation group：0.880±0.016，0.796±0.015；control group：0.481±0.007，0.474±0.021；inhibition group：0.207±0.014，0.167±0.005（P=0.000）. ③Protein expressions of TGF-β1，α-SMA，TGF-β-R1，Jagged1，Smad2/3，p-Smad2/3 were significantly upregulated in activation group and significantly downregulated in inhi－bition group. Activation group：1.076±0.063，1.180±0.138，1.192±0.142，1.582±0.247，2.057±0.114，2.088±0.120；control group：0.689±0.022，0.654±0.061，0.710±0.380，0.817±0.116，1.203±0.105，1.168±0.107；inhibition group：0.374±0.011，0.359±0.012，0.378±0.069，0.364±0.059，0.664±0.063，0.573±0.120（P=0.000）. The results above showed that there was significantly statistical difference between three groups（P<0.05）. Conclusion：TGF-β1 activates mouse hepatic stellate cells and regulate their bio－logical activities via Jagged1/Notch.

关键词

肝星状细胞转化生长因子-β1Jagged1

Keywords

hepatic stellate cellstransforming growth factor β1Jagged1

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