The Effects Of Ligustrazine And Simvastatin On High Glueose-induced T-PA And PAI-1 Expressions In Cultured Human Peritoneal Mesothelial Cells

Background Peritoneal dialysis (PD) is a form of renal replacement treatment, which has the advantages of better protection of residual renal function, better filtration effect for middle molecular substance and treating themselves at home. In recent years, PD gradually become the first choice of alternative treatment in such patients who are at end stage renal disease. But along with the continuous peritoneal dialysis, peritoneal fibrosis appeared gradually and worse steadily. Peritoneal fibrosis is a common complication of peritoneal dialysis, influence the effects of metabolic products and water by ultrafiltration, result in decreaseing the removal efficiency of creatinine, urea and guanidine toxin. Patients eventually drop out of peritoneal dialysis and choose to other replacement therapy. With the extracellular matrix (ECM) deposition, peritoneal vascular hyperplasia and damage of peritoneal mesothelial cells, peritoneal fibrosis aggravated gradually. At this time, studies have shown that High glucose concentration、low pH、high osmotic pressure and glucose degradation products in peritoneal dialysis fluid can induce the secretion of fibrogenic cytokines such as TGF-β,FGF, VEGF etc. Along with deposition of extracellular matrix, angiogenesis, peritoneal fibrosis occurs and increases. Previous study showed that, t-PA and PAI-1 are important cytokines, whose secretion and expression play an important role in peritoneal fibrosis process and influence the occurrence and development of peritoneal fibrosisObjective The research attempts to probe into the impacts of ligustrazine and simvastatin on high glucose-induced t-PA and PAI-1 secretion and expressions in HPMCs.Methods HPMCs were separated from human omentum through trypsinization and serial subcultivation. After concurrent of cells growth, HPMCs were distributed into normal control group (group N), high glucose-induced group(group H,2.5% glucose), high glucose-induced plus low-dose Simvastatin (group S1,2.5μmol/L Simvastatin), high glucose-induced plus mid-dose Simvastatin (group S2,5μmol/L Simvastatin), high glucose-induced plus high-dose Simvastatin (group S3,10μmol/L Simvastatin); high glucose-induced plus low-dose Ligustrazine(group T1,10mg/L Ligustrazine),. high glucose-induced plus mid-dose Ligustrazine(group T2,20mg/L Ligustrazine),. high glucose-induced plus high-dose Ligustrazine(group T3,40mg/L Ligustrazine),. MTT assay was used to measure the activity of HPMCs. Semi-quantitative RT-PCR was used to measure the mRNA expressions of t-PA and PAI-1 in HPMCs. Protein levels of t-PA and PAI-1 in culture supernatants were measured by ELISA. Cells protein concentration measured by BCA was to adjust the ELISA test results.Results①To contrast between the group N and group H, high glucose was able to depress cell viability (P<0.01) and the expression of t-PA in HPMCs significantly, meanwhile inerease the expression of PAI-1.②Compared with group H, Simvastatin can significantly improve the activity of HPMCs induced by high glucose(P<0.01). Otherwise, Simvastatin significantly reduced PAI-1 and raised t-PA expressions in HPMCs in a dose-dependent methods both in protein and gene levels (P<0.01).③Compared with group H,Ligustrazine also significantly ameliorate the viability of HPMCs in the intervention of high glucose(P<0.01).Above all, Ligustrazine markedly depressed PAI-1 expressions and increased t-PA in HPMCs in a dose-dependent manner both in protein and gene levels (P<0.01).Conclusion Simvastatin and Ligustrazine can depress high glucose-induced PAI-1 and promote t-PA expressions and improve the viability of HPMCs in the intervention of high glucose in vitro.This can confirm that Simvastatin and Ligustrazine play an important role in lessening the accumulation of ECM in HPMCs, meanwhile defending and/or postponing the procedure of peritoneal fibrosis in patients of continuous ambulatory peritoneal dialysis.