| ObjectiveTo study the renal expressions of NF-κB and MCP-1 in DN rats and the effects of sulodexide on it.MethodsFirst, selected 10 from the healthy male Wistar rats randomly, set to the normal control group (NC group), other 40 rats feed high sucrose and high fat food for one months, then, the diabetes mellitus model of Wistar rats received a single intraperitoneal injection of STZ dissolved in 0.1mol/L citrate buffer solution (PH 4.2) at a low dose of 35mg/kg. The rats in NC group were only received an injection of an equal volume of 0.1mol/L citrate buffer solution. The model of diabetes was considered to be successful when the blood glucose was≥16.7mmol/L after 72 hours of the injection. Then the forty successful diabetic model rats were randomly divided into four groups: diabetic model group (DM group, n=10), sulodexide treated group (DMS group, n=10), valsartan treated group (DMV group, n=10) and sulodexide combined with valsartan treated group (DMS+V group, n=10). The rats in the DMS,the DMV,the DMS+V group were treated intramuscularly with sulodexide (10mg.kg-1.d-1),orally with valstartan (10mg.kg-1.d-1),intramuscularly with sulodexide (10mg.kg-1.d-1) and orally with valsartan (10mg.kg-1.d-1), respectively. The rats of NC and DM group were terated intramuscularly with normal saline (1ml.d-1). After 12 weeks of drug administration, blood glucose (BG),triglyceride (TG),total cholesterol (TC),serum creatinine (Scr),urea nitrogen (BUN),urinary creatinine were measured routely; 24-hour urinary albumin excretion (UAE) was measured by nephelometry; the kidney hypertrophy index and the creatinine clearance (Ccr) were calculated. The expression of NF-κBP65,MCP-1 in kidney were detected by immunohistochemical method. HE staining was performed on renal tissue for light microscopy examination of mean glomerular volume (MGV).Results1. At the end of experimentation (after 12 weeks of drug administration), compared with NC group, DM group,DMS group,DMV group and DMS+V group had significant elevated BG,TG and TC levels (P<0.01); there were no significant difference of BG,TG,TC level among DM group,DMS group,DMV group and DMS+V group (P>0.05).2. Compared with NC group, DM group had significant increased UAE,Scr,BUN and kindey hypertrophy index level(P<0.01), and had significant decreased Ccr level(P<0.01); UAE,Scr,BUN and kindey hypertrophy index were significantly lower in DMS,DMV and DMS+V group than in DM group (P<0.01 or P<0.05), the Ccr was significantly higher in DMS,DMV and DMS+V group than in DM group(P<0.01, respectively); however, there were still statistical differences between NC group and every therapy groups (P<0.01 or P<0.05). UAE,Scr,BUN and kindey hypertrophy index were significantly lower in DMS+V group than in DMS and DMV group(P<0.01 or P<0.05),and the Ccr was significantly higher in DMS+V group than in DMS group and DMV group(P<0.01). There was no significant difference between DMS group and DMV group(P>0.05).3. Morphology changes were examined under light microscope. In the DM group,glomerulus has mesangial cell ground substance hyperplasia, mesangial region widen. Compared with NC group, MGV of DM group and every therapy groups were significantly increased (P<0.01 or P<0.05). Compared with DM group, every therapy groups reduced degree of renal pathological morphology, MGV of every therapy groups were significantly reduced (P<0.01 or P<0.05), and the group DMS+V reduced obviously. Compared with DMS,DMV group, MGV of DMS+V group was significntly reduced (P<0.01 or P<0.05). The MGV of DMV group was smaller than the DMS group, but there was no significant difference between the two groups (P>0.05).4. The immunohistochemical staining results showed that the renal expression of NF-κBP65 and MCP-1 increased remarkable in group DM compared with NC group (P<0.01). Compared with DM group, the renal expression of NF-κBP65,MCP-1 were significantly reduced in every therapy groups (P<0.01), and the DMS+V group reduced most obviously, but still higher than the NC group (P<0.01). The expression of NF-κBP65,MCP-1 was lower in DMS+V group than in DMS,DMV group (P<0.01 or P<0.05). There was no significant difference between DMS and DMV group (P>0.05).5. The correlation analysis results showed that renal NF-κBP65 expression was positively correlated with UAE and renal MCP-1 expression (r=0.584,P<0.01; r=0.902,P<0.01, respectively); renal NF-κBP65 expression was negatively correlated with Ccr (r=-0.796,P<0.01). The ranal MCP-1 expression also had positive correlation with UAE (r=0.680,P<0.01); and had negative correlation with Ccr (r=-0.740,P<0.01).Conclusions1. There were dysglycemia,lipodystrophia and more urinary albumin excretion in diabetic rats. And the diabetic rats also had sever renal functional and structural impairment. Meanwhile, the renal expression of NF-κBP65,MCP-1 in diabetic rats were significantly increased. This prompt that the abnormal expression of inflammatory factors NF-κB and MCP-1 may play an important role in the development and progression of diabetic nephropathy.2. Sulodexide can reduce the level of UAE of diabetic rats, improve the kidney function, attenuate the renal injury, decrease the renal expression of NF-κBP65 and MCP-1, and the mechanism of this renoprotection has no relative with the regulation of glucose and fat metabolism. This suggest that sulodexide possibly can retard the progression of the renal functional and structural injury through reducing the renal NF-κBP65 and MCP-1 expression, develop the effects of renoprotection and that is independent of glucose and fat metobolism regulation. Valsartan has the same effects in protecting kidney with sulodexide, and the combination of the two drugs has additive effects.3.The renal expression of NF-κBP65 was positively correlated with UAE, negatively correlated with Ccr. The renal expression of MCP-1 was also positively correlated with UAE, negatively correlated with Ccr. This prompt that the abnormal expressions of NF-κBP65 and MCP-1 are closely related to the dysfunction of kidney in DM rats. |
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