Experimental Study On Expression Change Of Inflammatory Cytokines And Protein Kinase C By Ligustrazine Combined With Benazepil Tablets To The Renal Interstitium In The GK Rats

Objective: To observe ligustrazine combined with benazepil tablets _prevention and cure in expression level of TGFβ1,MCP-1,TGFβ1mRNA in GK rats nephridial tissue tentatively and influences renal protein kinase C(PKC) activity and expression of PKCβisofoims in diabetic rat. To evalute the possible mechanisms of effects on diabetic neuropathy (DN) on earlier period. To probe into the connection between diabetic neuropathy and inflammatory to open out the possible mechanisms of DN for appraising and diagnosing new quideline on diabetic neuropathy on earlier period.Methods:Forty GK male rats were randomly divided into four groups:10 every groups, respectively for model group, ligustrazine group, benazepil group and ligustrazine combined with benazepil group. Drench with NS 5ml·kg-1·d-1,benazepill.6mg·kg-1·d-1,ligustrazine 150mg·kg-1·d-1,ligustrazine combined with benazepil (150mg·kg-1·d-1+1.6mg·kg-1·d-1) to rats respectively, according to the group in 16 weeks. Puting big mouses into metabolism cages in the 8th week, the 12th week and the 16th week, collecting urine to determine quantitation of Urine protein in 24 hours. Treat for 16 weeks, anaesthetize the big mouse by 3% pentobarbital,then cut open the belly of rats and pick its kidney, The mRNA levels of TGF-β1 in renal cortex were measured by reverse transcription polymerase chain reaction(RT-PCR),examine TGF-β1,MCP-1 content in renal organize by means of by means of immunohistochemistry, The PKC activity was measured by[3H] phorbol 12,13-dibutyrate([3H] PDBu) binding assay. The expression of PKCI andⅡisoforms in glomerulus was assessed by immunohistochemistry. Result:Compared with other group, the blood glucose of model group rats increase obviously (P<0.05), quantitation of 24h Urine protein rise obviously (P<0.05), degrade the express level of TGF-β1,MCP-1,TGFβ1mRNA and CPKβin renal organizes increase obviously (P<0.05,或p<0.01); ligustrazine combined with benazepil tablets can protect the kidney for lowerring blood glucose,TG and TC, degrading TGFβ-1,MCP-1,TGFβ1 mRNA express level in renal organize, decreasing serum albumin,blaood urea mitrogen and increasing creatinine clearance,at the same time, reducing Urine protein level.[3H] PDBu binding assay of group model showed a significant increase in memberane associated protein Icinase C (mPKC) activity and a significant decrease in cytosol associated protein kinase C (cPKC) activity in venal cortex compared with group normal(P<0.01); control group showed asignificant increase in cPKC activity and a significant decrease in mPKC activity compared with group DM (P<0.01); No significant difference was found between control group in renal cortical PKC activity (P>0.05). Immunohistochemistry showed a significant increase in PKCβ1 expression in glomerulus in group model,compared with in control group (P<0.01); PKCβⅠexpression in glomerulus in control group was significantly lower than in group model (P<0.01).The control group showed nodifference in glomemlar expression of PKCβⅠand renal cortical expression of PKCβⅠand CPKβⅡisoforms (P>0.05).Conclusion:ligustrazine combined with benazepil can reduce great mouse blood glucose level of diabetic rats and improve nephric tubule pathological change, moreover; it can play its roles depend on resisting inflammation and anticoagulatory function.One of the possible mechanisms of DN is to degrade TGF-β1,MCP-1 and TGFβ1mRNA and at least partially via the inhibition of PKC activation and PKCβisoform expression in renal cortex.