摘要
OBJECTIVE： To explore the function of Tangnai- kang （TNK） in the prevention and treatment of re- nal interstitial fibrosis through transdifferentiation of the human renal tubular epithelial cell line HK-2 induced bytransforming growth factor-β1 （TGF-β1）. METHODS： HK-2 cells cultured in dulbecco＇s modi- fied eagle medium/F12 （1：1） with 10% fetal calf se- rum were divided into six groups： blank control group, TGF-β1 group （TGF-β1 10 ng/mL）, serum con- trol group （TGF-β1 10 ng/mL ＋ 10% serum）, treat- ment group 1 （TGF-β1 10 ng/mL ＋ 5% TNK serum）, treatment group 2 （TGF-β1 10 ng/mL＋10% TNK se-rum）, and treatment group 3 （TGF-β1 10 ng/mL＋ 20% TNK serum）. Cell proliferation was detected by 4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazoliu m bromide assay. Expression of a-smooth muscle ac- tin （a-SMA） and E-cadherin were observed by im- munohistochemical assay. The contents of collagen Ⅰ （Col Ⅰ）, collagen Ⅲ（ColⅢ）, and fibronectin （FN） in the culture medium supernatant were detected by ELISA. RESULTS： E-cadherin was expressed and α-SMA was not expressed in normal HK-2 cells. In HK-2 cells cultured with TGF-β1, α-SMA expression signifi- cantly increased, HK-2 cells significantly proliferat- ed, and secretion of Col Ⅰ, Col Ⅲ, and FN significantly increased compared with the blank control group （all P〈0.05）. In the HK-2 cells cultured with TGF-β1 and TNK serum, the expression of α-SMA signifi- cantly decreased, the expression of E-cadherin sig- nificantly increased, and the cell proliferation and the secretion of Col Ⅰ, Col Ⅲ and FN were significant- ly inhibited compared with the TGF-β1 group （all P〈 0.05. CONCLUSION： TNK can inhibit cell proliferation and reduce secretion of Col Ⅰ, Col Ⅲ, and FN.This in- dicates that TNK can inhibit transdifferentiation of human renal tubular epithelial cells induced by TGF-β1, with the effect of preventing and treating renal interstitial fibrosis.
OBJECTIVE: To explore the function of Tangnaikang (TNK) in the prevention and treatment of renal interstitial fibrosis through transdifferentiation of the human renal tubular epithelial cell line HK-2 induced by transforming growth factor-β1(TGF-β1).METHODS: HK-2 cells cultured in dulbecco's modified eagle medium/F12 (1∶1) with 10% fetal calf serum were divided into six groups: blank control group, TGF-β1 group (TGF-β1 10 ng/mL), serum control group (TGF-β1 10 ng/mL+10% serum), treatment group 1 (TGF-β1 10 ng/mL+ 5% TNK serum),treatment group 2 (TGF-β1 10 ng/mL+10% TNK serum), and treatment group 3 (TGF-β1 10 ng/mL+20% TNK serum). Cell proliferation was detected by 4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Expression of α-smooth muscle actin (α-SMA) and E-cadherin were observed by immunohistochemical assay. The contents of collagen Ⅰ (Col Ⅰ), collagen Ⅲ (Col Ⅲ), and fibronectin (FN) in the culture medium supernatant were detected by ELISA.RESULTS: E-cadherin was expressed and α-SMA was not expressed in normal HK-2 cells. In HK-2 cells cultured with TGF-β1,α-SMA expression significantly increased, HK-2 cells significantly proliferated, and secretion of Col Ⅰ, Col Ⅲ, and FN significantly increased compared with the blank control group (all P<0.05). In the HK-2 cells cultured with TGF-β1 and TNK serum, the expression of α-SMA significantly decreased, the expression of E-cadherin significantly increased, and the cell proliferation and the secretion of Col Ⅰ, Col Ⅲ and FN were significantly inhibited compared with the TGF-β1 group (all P<0.05).CONCLUSION: TNK can inhibit cell proliferation and reduce secretion of Col Ⅰ, Col Ⅲ, and FN.This indicates that TNK can inhibit transdifferentiation of human renal tubular epithelial cells induced by TGF-β1, with the effect of preventing and treating renal interstitial fibrosis.
基金
Supported by the National Natural Science Fund(30973909)
Innovation Group Items of Beijing University of Traditional Chinese Medicine(No.2011-CXTD-19)
