Inhibitory Effect of Resveratrol on LPS-induced Glomerular Mesangial Cells Proliferation and TGF-β1 Expression via Sphingosine Kinase 1 Pathway / 中国结合医学杂志

OBJECTIVE@#To elucidate the renoprotective effect of resveratrol (RSV) on sphingosine kinase 1 (SphK1) signaling pathway and expression of its downstream molecules including activator protein 1 (AP-1) and transformation growth factor-β1 (TGF-β1) in lipopolysaccharide (LPS)-induced glomerular mesangial cells (GMCs).@*METHODS@#The rat GMCs line (HBZY-1) were cultured and randomly divided into 5 groups, including control, LPS (100 ng/mL), and 5, 10, 20 µmol/L RSV-treated groups. In addition, SphK1 inhibitor (SK-II) was used as positive control. GMCs were pretreated with RSV for 2 h and treated with LPS for another 24 h. GMCs proliferation was determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. The proteins expression of SphK1, p-c-Jun and TGF-β1 in GMCs were detected by Western blot, and DNA-binding activity of AP-1 was performed by electrophoretic mobility shift assay (EMSA). The binding activity between RSV and SphK1 protein was detected by AutoDock Vina and visualized by Discovery Studio 2016.@*RESULTS@#LPS could obviously stimulate GMCs proliferation, elevate SphK1, p-c-Jun and TGF-β1 expression levels and increase the DNA-binding activity of AP-1 (P<0.05 or P<0.01), whereas these effects were significantly blocked by RSV pretreatment. It was also suggested that the effect of RSV was similar to SK-II (P>0.05). Moreover, RSV exhibited good binding affinity towards SphK1, with docking scores of -8.1 kcal/moL and formed hydrogen bonds with ASP-178 and LEU-268 in SphK1.@*CONCLUSION@#RSV inhibited LPS-induced GMCs proliferation and TGF-β1 expression, which may be independent of its hypoglycemic effect on preventing the development of mesangial cell fibrosis and closely related to the direct inhibition of SphK1 pathway.

Effect of Bushen Tongluo Prescription on TGF-β1/Smads Pathway in Pulmonary Fibrosis Rats / 中国实验方剂学杂志

Objective:To observe the effect of Bushen Tongluo prescription on the pathogenesis of lung fibrosis induced by bleomycin and the transforming growth factor β1(TGF-β1)/Smads pathway protein in rats,and reveal the mechanism of this formula in inhibiting pulmonary fibrosis. Method:Totally 48 male SD rats were randomly divided into five groups:normal group(8 rats),model group(10 rats),high and low-dose Bushen Tongluo group(10 rats)and hydrocortisone sodium succinate group(10 rats). The model of pulmonary fibrosis was made with bleomycin except for normal group.Since day 3 after surgery,14.2,7.1 g·kg-1 Bushen Tongluo prescription were given by ig in high and low-dose Bushen Tongluo groups,and 4.58 g·kg-1 hydrocortisone sodium succinate was given by gavage(ig) in normal group and model group.All of the rats were put to death after 28 days.The contents of hydroxyproline (HYP),hyaluronic acid (HA) and laminin (LN) were measured after the death of the rats.Hematoxylin-eosin staining (HE) and Masson tricolor staining were used to observe the pathological changes of lung tissue. Real-time PCR and Western blot were used to observe the changes of mRNA and protein expressions of lung tissues TGF-β1, Smad2, Smad3, Smad7 in each group. Result:Compared with normal group, the level of pulmonary fibrosis was more significant in model group, and the serum HYP,HA and LN were increased obviously(PPβ1, Smad2,Smad3 proteins and mRNA in lung tissues were increased obviously(PPPPPβ1, Smad2, Smad3 protein and mRNA were decreased obviously(PPConclusion:Bushen Tongluo prescription can effectively improve the pathological process of pulmonary fibrosis, and the mechanism may be related to the regulation of TGF-β1/Smads pathway.

Effects of resveratrol on cell proliferation in glomerular mesangial cells cultured under high glucose by inhibiting SphK1/AP-1 signal pathway / 中国药理学通报

Aim To observe the expression of FN and TGF-β1 in the glomerular mesangial cells induced by high-glucose after the intervention of resveratrol, and further discuss its influence on SphK1/AP-1 signaling pathway. Methods The rat glomerular mesangial cells induced by high glucose were used to observe the effects of resveratrol on cell proliferation after interven-tion. The survival vitality and proliferation of glomeru-lar mesangial cells were determined by MTT, and then FN, TGF-β1 and SphK1 protein expression were deter-mined by Western blot. Also, AP-1 activity was deter- mined by EMSA assay. Results Resveratrol could obviously inhibit the proliferation of high glucose-in-duced glomerular mesangial cells, lower SphK1 expres-sion, inhibit AP-1 activity and thus inhibit the expres-sion of FN, TGF-β1. Conclusions Resveratrol inhib-its the proliferation of high glucose-induced glomerular mesangial cells, which may be closely related to the in-hibition of SphK1/AP-1 signaling pathway.

Mechanism and clinical significance of TGF-β1 signal transduction pathway regulated by VEGF gene in nasopharyngeal carcinoma metastasis / 实用医学杂志

Objective To explore the relationship between the expression of VEGF and TGF-β1 in naso-pharyngeal carcinoma and the clinical characteristics and prognosis of nasopharyngeal carcinoma patients. Meth-ods The expression of VEGF and TGF-β1 in the local middle-late nasopharyngeal carcinoma tissue of 96 patients was detected using immunohistochemistry.The correlation and the prognosis analysis were analyzed in combination with their clinical characteristics and prognosis. Results Expression rate of VEGF was 66.7% and that of TGF-β 1 53.1%.The expression of VEGF and TGF-β1 were positively correlated in the advanced nasopharyngeal carcino-ma.Both VEGF and TGF-β1 had significant correlation with lymphatic metastasis.The effects of VEGF and TGF-β1 on MFS were analyzed by Cox single factor and the results showed that VEGF positive was an adverse prognostic factor(P<0.05).The effects of age,sex,lymphatic metastasis,local invasion,VEGF and TGF-β1 on MFS were analyzed with Cox multifactor analysis and the results showed that VEGF positive was an adverse prognostic factor (P < 0.01). Conclusion The detection of VEGF and TGF-β1 protein in nasopharyngeal carcinoma may help to predict the metastasis of nasopharyngeal carcinoma.

Correlation analysis of Treg cells and TGF-β1, IL-10 levels in a rat experimental liver carcinoma model / 中华普通外科杂志

Objective To study the relationship between Treg cell numbers and level of TGF-β1,IL-10 in the immune microenvironment of rat liver cancer.Method 40 male Wistar rats were randomly divided into three groups,liver cancer model group (n =20),saline control group (n =10) and blank control group (n =10).Liver cancer model was established by intraperitoneal injection of DEN (100 mg/kg).Percentage of Treg cells in cancer tissues was detected by fiow cytometry and the expression level of TGF-β1 and IL-10 by immunohistochemical SABC.Results The ratio of Treg cells in CD4 + T cells was 14.32％ ± 4.84％ in liver cancer tissues,significantly higher than that in the blank control group 5.64％ ±6.10％ and the saline control group 7.95％ ±3.55％.The difference was statistically significant (F =211.279,P ＜ 0.05).The expression level of TGF-β1 and IL-10 in cancer tissues was significantly higher than the blank control group and the saline control group (FTGF-β1 =250.740,FIL-10 =152.744,all P ＜0.05).The number of Treg cells was positively correlated with TGF-β1,IL-10 level (P ＜ 0.05).Conclusions TGF-β1,IL-10 and Treg cells significantly increased in rat experimental liver cancer tissues,and there was positive correlation between Treg cells,and TGF-β1 and IL-10 in liver cancer tissues.

Change of serum associated factors in patients with combined pulmonary fibrosis and emphysema / 中国综合临床

Objective To investigate the levels of transformation growth factor-β1 (TGF-β1),vascular endothelial growth factor (VEGF) and insulin growth factor-Ⅰ (IGF-Ⅰ) role in the pathogenesis of serum of patients with combined pulmonary fibrosis and emphysema (CPFE).Methods Recruited 20 patients with CPFE,40 cases with idiopathic pulmonary interstitial fibrosis (IPF) and 40 cases with emphysema who were admitted to our hospital during July 2011 to February 2012.Serum levels of TGF-β1,VEGF and IGF-Ⅰ were detected by ABC-ELISA.Results Serum levels of TGF-β1 and IGF-Ⅰ were significantly higher in patients with CPFE and IPF than these in patients with emphysema (TGF-β1:(160.73 ± 40.62) ng/L vs.(167.35 ± 42.82) ng/L vs.(128.75 ±35.77) ng/L; IGF-Ⅰ:(179.65 ±60.73) ng/L vs.(192.32 ±58.75) ng/L vs.(148.73 ±49.67) ng/L,P ＜ 0.05 or P ＜ 0.01).The IPF group had significantly higher serum level of VEGF than the emphysema group ((506.12 ±82.37) ng/L vs.(437.31 ±62.58) ng/L,P ＜0.01).Serum levels of TGF-β1 and IGF-Ⅰ in CPFE and emphysema groups were positively correlated (r =0.885,0.918 respectively,P ＜ 0.01).Smokers in the CPFE group had significandy lower level of serum VEGF than those who did no smoke ((406.19 ± 66.94) ng/L vs.(482.88 ± 79.91) ng/L,t =-2.287,P =0.035).Conclusion Serum level of VEGF increased significantly in the IPF group,suggesting the participation of VEGF in pulmonary interstitial fibrosis.IGF-Ⅰ involved in the pathogenesis of pulmonary interstitial fibrosis.TGF-β1 and IGF-Ⅰ have a positive linear regressive relationship,which indicates that they may work synergistically in the process of the fibrosis.

Effects of high glucose on the proliferation of HSC and expressions of TGF-?_1,PDGF and PCⅢ in HSCs / 西安交通大学学报(医学版)

Objective To investigate the effects of high glucose on the proliferation of hepatic stellate cell-T6(HSC-T6) and the expressions of transformation growth factor-?1(TGF-?1),platelet-derived growth factor(PDGF) and precollagen Ⅲ(PCⅢ).Methods Based on glucose groups of different concentration,we observed the proliferation of HSC in 30min-16h time period,and used the methods of immunohistochemistry and radioimmunoassay to measure the expressions of TGF-?1,PDGF and PCⅢ in the supernatant at 48h and 72h.Results In 2-16h time period,the proliferation of HSC was increased stepwise over time in 1500-6000mg/L group,and was more visible in 4500-6000mg/L group.The expressions of TGF-?1 and PDGF increased at 48h,and the expression of PCⅢ in the supernatant increased at 72h in 4500-6000mg/L group compared with that in glucose control and hypertonic control groups.Conclusion High glucose can promote hepatic fibrosis by stimulating the expressions of TGF-?1,PDGF and PCⅢ in HSC-T6.