Effects Of Hepatocyte Growth Factor On Peritoneal Fibrosis Induced By High Glucose

The Effects of Hepatocyte Growth Factor on Peritoneal Fibrosis Induced by High GlucoseBackground and Objective:Peritoneal dialysis(PD) is a major treatment modality for end-stage renal failure.Nevertheless,the peritoneal membrane exposed continuously to bioincompatible PD solutions could be injured to fibrosis,which lead patients to discontinue PD.EMT of HPMCs induced by TGF-β₁ which is up-regulate in HG milieu is a key process in the initiation and perpetuation of peritoneal fibrosis.It has been reported that the reciprocal imbalance between TGF-β₁ and HGF is closely involved in the progression of tissue fibrosis.Some studies have indicated that HGF is an endogenous, antifibrotic factor.Therefor,we try to investigate whether exogenous HGF could influence the up-regulation of TGF-β₁ in HPMCs cultured with HG,and prevent EMT of HPMCs, decreases the synthesis of ECM.Our final aim is to understand the role of HGF on the prevention and treatment of peritoneal fibrosis induced by high-glucose in the patients with peritoneal dialysis.Method:HPMCs were isolated from specimens of the omentum in nonuremic patients by enzymatic disaggregatio.HPMCs were identified by phase-contrast microscopy according to the morphologic criteria and by the immunohistochemistry.HPMCs from the third passages were used in this experiment.After sub-confluent culture HPMCs were incubated in serum-free medium for 24 hours,HPMCs were exposed to different concentrations of glucose(5.5,30,60mmol/L D-glucose;60mmol/L mannitol),and the HPMCs cultured with HG(60mmol/L D-glucose) were treated with rhHGF(25,50,100ng/ml) for 48h.The level of TGF-β₁ and FN protein in the supernatant were measured by ELISA assay.α-SMA in HPMCs was tested by immuocytochemistry.The expressions of TGF-β₁,FN,α-SMA mRNA in HPMCs were detected by Real-Time PCR.Result:1.The cultured HPMCs were spindly at first.A week later,they connected with each other and took on a slabstone-like appearance.Immunohistochemistry analysis showed positive immunoreactivity of vimentin and cytokeratin in these cells.2.The expression ofα-SMA,TGF-β₁,FN mRNA and protein increased significinely in HPMCs cultured with HG(P＜0.05),but did not increase in HPMCs cultured with mannitol. Significant positive correlation was found between the concentrations of glucose and the expression ofα-SMA,TGF-β₁,FN(P＜0.01).3.Up-regulation of the expression ofα-SMA,TGF-β₁,FN mRNA and protein induced by HG was suppressed by HGF. Significant negative correlation was found between the concentrations of HGF and the expression ofα-SMA,TGF-β₁,FN(P＜0.01).Conclusion:1.HG can induce the expression of TGF-β₁,and EMT of cells,promote the synthesis of ECM in HPMCs.2.HGF can suppress the up-regulation of TGF-β₁ induced by HG,prevent EMT of cells.,and decrease synthesis of ECM in HPMCs.These results suggest that HGF could inhibit HG-inducing peritoneal fibrosis of the patients with PD.