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400038 重庆,陆军军医大学(第三军医大学)第一附属医院全军消化病研究所
[彭贵勇,E-mail:pgy63@163.comPENG Guiyong, E-mail: pgy63@163.com
收稿:2017-10-12,
修回:2017-12-11,
纸质出版:2018-03-15
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王凌, 李敏, 贾益君, 等. 转化生长因子β1对L929细胞增殖转分化及细胞外基质代谢的影响[J]. 第三军医大学学报, 2018,40(5):367-371, 379.
Ling WANG, Min LI, Yijun JIA, et al. Effects of TGF-β1 on transdifferentiation and extracellular matrix synthesis in L929 cells[J]. Journal of Third Military Medical University, 2018, 40(5): 367-371, 379.
王凌, 李敏, 贾益君, 等. 转化生长因子β1对L929细胞增殖转分化及细胞外基质代谢的影响[J]. 第三军医大学学报, 2018,40(5):367-371, 379. DOI: 10.16016/j.1000-5404.201710068.
Ling WANG, Min LI, Yijun JIA, et al. Effects of TGF-β1 on transdifferentiation and extracellular matrix synthesis in L929 cells[J]. Journal of Third Military Medical University, 2018, 40(5): 367-371, 379. DOI: 10.16016/j.1000-5404.201710068.
目的
2
探讨转化生长因子β1(transforming growth factor-β1,TGF-β1)对L929细胞增殖的影响,为体外研究预防食管内镜黏膜下剥离术(endoscopic submucosal dissection,ESD)术后狭窄提供理论基础。
方法
2
实验分为两组,①空白对照组:L929细胞不做处理;②TGF-β1组,用10 ng/mL TGF-β1持续刺激L929细胞,依据刺激时间不同又分为24、48、72、96、120 h 5个亚组。观察各组对细胞增殖的影响。用CCK-8法检测细胞增殖,qPCR检测α-SMA及Ⅰ型前胶原、Ⅲ型前胶原、MMP1、TIMP1 mRNA表达,Western blot检测α-SMA、MMP1、TIMP1蛋白水平,ELISA检测Ⅰ型前胶原、Ⅲ型前胶原合成。
结果
2
与空白对照组比较,TGF-β1组细胞增殖能力显著增高(
P
<
0.05),TGF-β1组α-SMA、Ⅰ型前胶原、Ⅲ型前胶原、MMP1、TIMP1 mRNA表达均显著升高(
P
<
0.05);TGF-β1刺激24 h组Ⅰ型前胶原合成增加(
P
<
0.05),TGF-β1组Ⅲ型前胶原合成增加(
P
<
0.05);TGF-β1组MMP1、TIMP1蛋白表达增加(
P
<
0.05)。
结论
2
TGF-β1能促进L929细胞增殖,并诱导L929细胞向肌成纤维细胞转分化,同时促进Ⅰ型前胶原、Ⅲ型前胶原以及MMP1、TIMP1合成。
Objective
2
To investigate the effect of TGF-β1 on the growth of L929 cells in order to found the basis for theoretical study on prevention of esophageal stricture after endoscopic submucosal dissection.
Methods
2
Experiments had two sets: a control group without any treatments and a TGF-β1 treatment group in which cells were treated with 10 ng/mL TGF-β1. According to the time of stimulation by TGF-β1
subgroups were set as 24
48
72
96 and 120 h treatment groups
the cell proliferation were measured with CCK-8 assay in every group of cells. qPCR was used to detect the mRNA expression of α -SMA and procollagen type Ⅰ (Col Ⅰ) and Ⅲ (Col Ⅲ)
matrix metalloproteinase 1 (MMP1)
tissue inhibitor of metalloprotein 1 (TIMP1). Western blotting was employed for protein levels of α -SMA
MMP1 and TIMP1. ELISA was adopted to measure the synthesis of Col Ⅰ and Col Ⅲ.
Results
2
Compared with the blank control cells
TGF-β1 treatment significantly promote cell proliferation (
P
<
0.05)
and enhanced the mRNA levels of α-SMA
Col Ⅰ
Col Ⅲ
MMP1 and TIMP1 in L929 cells (
P
<
0.05). TGF-β1 stimulation for 24 h increased the syntheses of Col Ⅰ (
P
<
0.05) and Col Ⅲ (
P
<
0.05)
and protein level of MMP1. The treatment for 96 and 120 h also enhanced the protein level of TIMP1 (
P
<
0.05).
Conclusion
2
TGF-β1 can promote the proliferation of L929 cells
induce the transformation of L929 cells into myofibroblasts
and improve the syntheses of Col Ⅰ
Col Ⅲ
MMP1 and TIMP1.
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