| Objective: 1. To establish the type 2 diabetic nephropathy rats model,exploring the correlation between the expression of Mannose-binding lectin(MBL) and inflammatory cytokine on rat’s kidney and Diabetic nephropathy. 2. Observing the effects of triptergium wilfordii on the expression of Mannose-Binding Lectin( MBL) and inflammatory cytokine in renal tissue of diabetic nephropathy rats, to explore the potential protective mechanism of triptergium wilfordii on diabetic nephropathy.Method:46 male SD rats were randomly divided into experimental group(36)and normal group(10), after 6 weeks high-suger and high-fat diet, the experimental group diabetes were injected into streptozocin(STZ) 40 mg.kg-1for DN rat model.The Rat models were randomly divided into diabetes mellitus model group(group DN) of 17,and tripterygium wilfordii treated diabetes mellitus group(group TW) of 18,that were treated with tripterygium wilfordii(10mg/Kg.d) for 8 weeks.After 16 weeks, the weight of each group,Kidney index,24 hours protein(24h U-TP), blood-glucose and biochemical indexes were measured; Partial nephridial tissues were treated with HE staining; Immunohistochemical were used to detect the expression of MBL-A,MASP2, IL-6,TNF-α protein;Real-time PCR was used to detect MBL1 and MASP2 m RNA expression in renal tissue;Western blot were used to detect the expression of MBL-A and MASP2 protein.Result: 1. After the end of the experiment, DN group and TW group rats were died 4 and 5 separately. The rats in group DN were depressed, unresponsive, reduced activity,significant weight loss, shaggy Brown without luster, polydipsia and polyuria, loose stool. TW intervention improved the situation. 2. Compared with NC group, the renal index,24 h UTP, BG, SCr, BUN of DN group were significantly increased, the body weight apparently losse(P<0.01); compared with DN group, the renal index in rats of TW group decreased(P < 0.05), 24 h UTP, SCr, BUN decreased significantly(P < 0.01), and body weight and BG have no obvious changed(P > 0.05). 3. Pathologic study showed: Compared with NC group,the glomerular volume of DN and TW group were significantly increased(P<0.01),interstitial inflammatory cell infiltration increased(P<0.05); compared with DN group, the TW group rats glomerular volume shrinking, renal interstitial infiltration of inflammatory cells decreased in number(P<0.05). Renal tissue immunohistochemical staining showed: compared with NC group, the expression of MBL-A, MASP2, IL-6, TNF-a protein significantly increased in the other two groups(P<0.01); compared with DN group,the expression of MBL-A,MASP2 protein of TW group decreased(P < 0.05), the expression of IL-6, TNF-a significantly reduced(P < 0.01). Real-Time PCR results suggest: Expression of MBL1, MASP2 m RNA in DN group were significantly increased compared with that of NC group(P<0.01); expression of MBL1, MASP2 m RNA in TW group was lower than in group DN(P<0.05). Western blot results indicated: the expression of MBL-A, MASP2 protein of DN group were significantly increased compared with that of NC group(P < 0.01); Compared with DN group, the expression of MASP2 protein in group TW decreased(P<0.05), and MBL-A protein was significantly decreased(P < 0.01).Conclusion: 1. The mannose binding-lectin pathway and inflammatory factors involved in kidney damage of diabetic rats; 2. Tripterygium wilfordii can protect the renal tissue of diabetic rats. It may be related to the inhibition of the lectin pathway of complement activation, reduced the expression of inflammatory cell exudation and inflammatory cytokines IL-6,TNF-a, thereby reducing the inflammatory immune response to renal injury. |
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