The Effect Of Ages On Trpc6 Expression In Glomerular Mesangial Cell

Background:Diabetes is increasing dramatically worldwide, and diabetic nephropathy (DN), the most common cause of end-stage, is one of the main microvascular complications, of which the disease mechanisms are extremely complicated, including advanced glycation end products (ACEs). ECM accumulation and GBM thickening are pathological characteristics of DN. Glomerular mesangial cell (GMC) is more active in the pathophysiology of DN. Calcium as a second messenger, involves in many biological functions, such as contraction, secretion, proliferation, and apoptosis in GMC.In recent years GMC proliferation has been researched accompanied with mesangial cell membrane ion channels open and close. Further researches of GMC membrane ion channel are of great significances for prevention and treatment of diabetic nephropathy. Transient receptor potential (TRP) is a newly discovered membrane cation channel, mainly through Ca2+. Transient receptor potential C (TRPC), the traditional-type TRP, is a key member of the TRP family. Recent studies show that TRPC6 associates with kidney disease, such as familial focal segmental glomerulosclerosis (FSGS), proteinuria, the reseach of which associates with DN is poor. We investigate whether AGEs affect TRPC6 location in GMC and TGF-β1 involves in it.Objective:To study whether the AGEs effect TRPC6 expression in GMC and TGF-β1 involves in it, further investigating AGEs in the pathogenesis of diabetic nephropathy.Methods:GMC of three generations were primarily cultured in RMPI 1640 supplemented with.20% fetal bovine serum, being grouped, respectivly treated with AGE-BSA,BSA, TGF-β1 for 24 hours,designing the control group, and then cells were collected, RNA and proteins extracted, Western blot, RT-PCR and immunofluorescence detecting TRPC6 location. Results:1. TRPC6 was detected in GMC with inmmunofluorescence, cell membrane mainly located.2. TRPC6 expression of different doses of AGE-BSA, BSA in GMC:BSA treated groups had no statistical significance (p> 0.05). AGE-BSA down-regulated TRPC6 location in the dose-dependent way, with statistical significance (p<0.05).3. AGE-BSA treated group attenuated TRPC6 appearance in the inmmun of luorescence way.4. Compared with normal group, TRPC6 mRNA and protein levels of TGF-β1 (10ng/ml)group decreased(p<0.05).5. Compared with normal group, Inmmunofluorescence showed the reduction of TRPC6 protein expression in TGF-β1 (10ng/ml) group.Conclusion:GMC expressed TRPC6, cell membrane mainly located; AGEs and TGF-β1 down-regulated TRPC6 protein, also AGEs increased TGF-β1 appearance in GMC, suggesting that AGEs down-regulated TRPC6 expression maybe partly by means of TGF-β1.