Objective:To investigate the effect of saponin from the seed of Litchi chinensis Sonn(SLS)on the growth and apoptosis of human kidney epithelial cells(HKC)cultured in high glucose.Methods:HKC were cultured in DMEM/F12...Objective:To investigate the effect of saponin from the seed of Litchi chinensis Sonn(SLS)on the growth and apoptosis of human kidney epithelial cells(HKC)cultured in high glucose.Methods:HKC were cultured in DMEM/F12 medium supplemented with 30 mmol/L glucose and treated with or without SLS.In the normal group,isometric DMEM/F12 medium with 5.5mmol/L glucose was added.The secretion of TGF-β1 and fibronectin(FN)were detected by ELISA.Cell apoptosis was detected by the method of Annexin V-FITC/PI double staining.Western blot was used to detect the level of suppressor of cytokine signaling-1(SOCS-1).Results:The result of ELISA showed that the secretion of TGF-β1 and FN was decreased in SLS groups compared with those in 30 mmol/L glucose treated group(P<0.05).There were more cells apoptosis in 30 mmol/L glucose treated group than that in the normal group(P<0.01).Compared with the 30 mmol/L glucose treated group,the apoptosis of HKC were significantly decreased in SLS groups(P<0.01).Western blot showed that the level of SOCS-1 in high glucose+SLS group was decreased(P<0.01),compared with the high glucose group.Conclusion:SLS can reduce the secretion of TGF-β1 and FN in HKC by reducing the deposition of extracellular matrix.SLS also significantly reduced the apoptosis of HKC by inhibiting the level of SOCS-1.These results suggest the roles of SLS in preventing the progress of glomerular sclerosis.展开更多
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 b...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.展开更多
Objective： The effects of hydraulic pressure on renal tubular epithelial-myofibroblast transdifferentiation （TEMT） were investigated. Methods： We applied hydraulic pressure （50 cmH2O） to normal rat kidney tubula...Objective： The effects of hydraulic pressure on renal tubular epithelial-myofibroblast transdifferentiation （TEMT） were investigated. Methods： We applied hydraulic pressure （50 cmH2O） to normal rat kidney tubular epithelial cells （NRK52E） for different durations. Furthermore, different pressure magnitudes were applied to cells. The morphology, cytoskeleton, and expression ofmyofibroblastic marker protein and transforming growth factor-β1 （TGF-β1） of NRK52E cells were examined. Results Disorganized actin filaments and formation of curling clusters in actin were seen in the cytoplasm of pressurized cells. We verified that de novo expression α-smooth muscle actin induced by pressure, which indicated TEMT, was dependent on both the magnitude and duration of pressure. TGF-β1 expression was significantly upregulated under certain conditions, which implies that the induction of TEMT by hydraulic pressure is related with TGF-β1. Conclusion： We illustrate for the first time that hydraulic pressure can induce TEMT in a pressure magnitude- and duration-dependent manner, and that this TEMT is accompanied by TGF-β1 secretion.展开更多
基金National Natural Science Foundation of China(No.81374025)the Education Department of Jilin Province“13th Five-Year”science and technology research project.
文摘Objective:To investigate the effect of saponin from the seed of Litchi chinensis Sonn(SLS)on the growth and apoptosis of human kidney epithelial cells(HKC)cultured in high glucose.Methods:HKC were cultured in DMEM/F12 medium supplemented with 30 mmol/L glucose and treated with or without SLS.In the normal group,isometric DMEM/F12 medium with 5.5mmol/L glucose was added.The secretion of TGF-β1 and fibronectin(FN)were detected by ELISA.Cell apoptosis was detected by the method of Annexin V-FITC/PI double staining.Western blot was used to detect the level of suppressor of cytokine signaling-1(SOCS-1).Results:The result of ELISA showed that the secretion of TGF-β1 and FN was decreased in SLS groups compared with those in 30 mmol/L glucose treated group(P<0.05).There were more cells apoptosis in 30 mmol/L glucose treated group than that in the normal group(P<0.01).Compared with the 30 mmol/L glucose treated group,the apoptosis of HKC were significantly decreased in SLS groups(P<0.01).Western blot showed that the level of SOCS-1 in high glucose+SLS group was decreased(P<0.01),compared with the high glucose group.Conclusion:SLS can reduce the secretion of TGF-β1 and FN in HKC by reducing the deposition of extracellular matrix.SLS also significantly reduced the apoptosis of HKC by inhibiting the level of SOCS-1.These results suggest the roles of SLS in preventing the progress of glomerular sclerosis.
基金Supported by the National Natural Science Fund(30973909)Innovation Group Items of Beijing University of Traditional Chinese Medicine(No.2011-CXTD-19)
文摘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.
基金Project (No. 2007CB947802) supported by National Basic Research Program of China
文摘Objective： The effects of hydraulic pressure on renal tubular epithelial-myofibroblast transdifferentiation （TEMT） were investigated. Methods： We applied hydraulic pressure （50 cmH2O） to normal rat kidney tubular epithelial cells （NRK52E） for different durations. Furthermore, different pressure magnitudes were applied to cells. The morphology, cytoskeleton, and expression ofmyofibroblastic marker protein and transforming growth factor-β1 （TGF-β1） of NRK52E cells were examined. Results Disorganized actin filaments and formation of curling clusters in actin were seen in the cytoplasm of pressurized cells. We verified that de novo expression α-smooth muscle actin induced by pressure, which indicated TEMT, was dependent on both the magnitude and duration of pressure. TGF-β1 expression was significantly upregulated under certain conditions, which implies that the induction of TEMT by hydraulic pressure is related with TGF-β1. Conclusion： We illustrate for the first time that hydraulic pressure can induce TEMT in a pressure magnitude- and duration-dependent manner, and that this TEMT is accompanied by TGF-β1 secretion.
