The Study Of The Expression Of Osteopontin And Its Acceptor Stimulated By High Concentration Glucose In Human Renal Tubular Epithelial Cell And The Intervention Of Mycophenolic Acid

Objective To investigate the expression of osteopontin (OPN) and its acceptor integrinα_v,integrinβ₁ and TGF-β₁ in the renal tubular epithelial cell stimulated by high concentration glucose,to discuss the effect of these cell factor in tubulointerstitial fibrosis,and make use of the new kind of immunosuppressant mycophenolic acid (MPA)to intervene,in order to find out its effect for several cell factors and the effect for diabetic nephropathy.Methods To cultivate the human renal tubular epithelial (HK-2) in vitro,divide the HK-2 into four groups by the difference of medium ,the group of A: the medium of low concentration glucose; B:the medium of high concentration glucose;C: the medium of high concentration glucose+10-3mmol/lMPA ; D: the medium of high concentration glucose+10-2mmol/lMPA,separately measure at 48 and 72 hours and evaluate the protein of OPN by Elisa ,OPN mRNA by RT-PCR and integrinα_v,integrinβ₁and TGF-β₁ by immunohistochemistry.Results after 24 hours the level of the express of OPN in group of the high glucose increase obviously than in group of the low glucose,and the level of OPN mRNA in tubular epithelial cell also obvious increase, P﹤0.05,and the level of OPN and its mRNA increase further with time,after 72 hours the level of OPN in group of the high glucose get 4.563 times by the level of OPN in the group of the low glucose,and the level of OPN mRNA in tubular epithelial cell in group of the high glucose get 3.141 times by the level of OPN mRNA in the group of the low glucose; The outcome of immunohistochemistry reveals after 48 hours the positive percent of integrinα_v in the group of high glucose (5.235±0.618) obviously increase than in the group of low glucose (0.686±0.262),the positive percent of TGF-β₁ in the group of high glucose (10.795±1.582)also obviously increase than in the group of low glucose (2.428±0.261),after 72 hours the positive percent of integrinβ₁ in the group of high glucose (10.228±1.282)obviously increase than the group of low glucose (4.365±0.328), P﹤0.05;after adding MPA,the high level of the cell factor in different group all descend to a certain extent, P﹤0.05,and there is the difference depending on the concentration between the group of 10-2mmol/lMPA and the group of 10-3mmol/lMPA, P﹤0.05;After adding the minim albumin ,the level of the cell factor have not increase obviously ;the correlation analysis reveals that OPN,integrinα_v,integrinβ₁and TGF-β₁ were positively correlated.Conclusions 1.The high concentration glucose can enhance the expression of OPN,integrinα_v,integrinβ₁,and these cell factors are positively correlated, it reveals that the abnormal OPN in diabetic nephropathy affect by connecting to integrinα_v and integrinβ₁.2.The high concentration glucose can increase the expression of TGF-β₁ , TGF-β₁and OPN,integrinα_v,integrinβ₁ are positively correlated, it reveals that all above cell factors affect all together in diabetic nephropathy.3. MPA can restrain the expression of the above cell factors, and different concentration has different effect,this can reveal that MPA can decrease the abnormal expression of the above cell factors in diabetic nephropathy, and it maybe have some good effect on diabetic nephropathy.