TGF-β1对人腹膜间皮细胞转分化影响的探讨

The effects of TGF-β1 on transdifferentiation of human peritoneal mesothelial cells

目的探讨TGF-β1刺激对人腹膜间皮细胞(HPMCs)转分化的影响机制。方法用5ng/m l TGF-β1刺激第三代HPMCs培养细胞,采用免疫组化法观察细胞内磷酸化Sm ad-2(p-Sm ad-2)的蛋白表达,W estern b lot法观察细胞内α-平滑肌肌动蛋白(α-SMA)、I型胶原(COL-1)和p-Sm ad-2的蛋白表达,ELISA观察上清液纤维连接蛋白(FN)的蛋白表达,以及RT-PCR观察α-SMA、FN和COL-1A1的mRNA表达水平。结果刺激组α-SMA、FN和COL-1A1的mRNA及α-SMA、FN、COL-1和p-Sm ad-2蛋白的表达显著增加,且均呈时间依从。结论TGF-β1能诱导HPMCs转分化为肌成纤维细胞,其可能的机制与激活了Sm ad信号通路有关。

Objective To explore the effects of TGF-β1 stimulation on the transdifferentiation of human peritoneal mesothelium cells （HPMCs）. Methods The greater omentum cells from healthy adults were used for primary culture and the third generation cells were stimulated hy 5ng/ml TGF-β1. Immunohistochemistry assay was used to detect the expression of p-Smad-2. Western hlot assay was used to detect the expressions of α- smooth muscle aetin （α-SMA）, collagen, type Ⅰ （COL-1） and p-Smad-2 in cells. The contents of fihronectin （FN） in supernatant were detected by EI,ISA, and the mRNA expressions of α-SMA, FN and COL-1 in cells were analyzed hy RT - PCR. Results The mRNA levels as well as their protein expressions of α-SMA, FN adn COL-1 in the treatment groups were. significantly increased than those of the control group, and the mRNA and protein expressions displayed the time dependence. Conclusion HPMCs can be transdifferentiated into nlynfilbrohlast under the effects of TGF-β1 and the mechanism may he related with the activation of Smad signal pathway in HPMCs.