The Expression Of RANTES And TGF-β In Tubule Epithelial Cells Under High Glucose Concentration And The Intervention Effect Of Mycophenolic Acid On Them

Objective:To investigate the expression of NF-κB,RANTES and TGF-β1,IV collagen in human tubule epithelial cells which incubated under the condition of high glucose,and the intervention effect of Mycophenolic acid on them,to discuss the protect's effect of Mycophenolate mofetil on the tubule epithelial cells under high glucose concentration.Methods:In vitro,incubate human tubule epithelial cells and divide into three groups according to different medium: (1)control group(glucose's concentration:5.5mmol/l) (2)high glucose group (glucose's concentration :25mmol/l) (3)MPA+high glucose intervention group(concentration of MPA: 10-5mol/l);measure the content of RNATES by ELISA,the expression of NF-κBmRNA by semi-quantity RT-PCR ,and observe the expression of TGF-β1,IV collagen by immunohistochemistry.Results: 1. HK-2 has its basal level of RANTES protein,under high glucose condition,the content of RNATES protein increases as time manner。After 48hs,the content of RNATES began to increase(P<0.05);after 72hs,the expression's increasing had the significant difference(P<0.01). RT-PCR examination showed the expression of NF-κBmRNA increase as time manner in high glucose group,which are significantly higher than that in control group at the same time(P<0.01),increase 1.11,2.07,2.41 times respectively after 24hs,48hs,72hs.2. The result of cell immunohistochemistry showed the expression of TGF-β1 and IV-C are mostly in the cell plasma , positive cell expression percentage increase significantly as time manner after stimulated by high glucose(P<0.01).3. RANTES expression of HK-2 reduced distinctly in MPA group than that in the high glucose group, RANTES expression reduced sharply after 48hs'and 72hs' intervention by MPA(P<0.05),the level of NF-κbmRNA were lower distinctly in MPA grou(P<0.01),and depressed 32.0%,35.4%,39.3% compared with high glucose group respectively after 24hs,48hs,72hs. After intervention by MPA, positive cell percentage of TGF-β1 and IV-C decreased distinctly compared with than in the high glucose group(P<0. 05).Conclusions:1.Expression of NF-κB,RANTES in HK-2 stimulating by high glucose is significantly higher than control group as time manner. 2. Expression of TGF-β1 and IV-C stimulating by high glucose is significantly higher than control group, and there was a positive correlation between them. 3. Expression of NF-κB,RANTES,TGF-β1,IV-C is depressed significantly by MPA,and MPA can play a protective role on tubule epithelial cells by restaining the expression of the above factors, which can offer experimental elements for the clinical application of mycophenolate mofetil ( MMF )on diabetic nephropathy.