| ObjectiveTaking the high-fat high-sugar diet to feed and intraperitoneal injection of streptozotocin SD rats as our research objects.Taking Tripterygium Wilfordii Polyglucoside (TWP) Tablets as the treatment drug.We observed the infiltration of macrophages an the expression of NF-κB、MCP-1、in SD rats’kidney,and discussed their effection on diabetic nephropathy and the possible mechanism. The effect of different dosages of TWP Tablets on the macrophages and the related cytokines in prevention and early treatment was also our reaserch purpose.Method136clean grade male SD rats were adopted as the research objects and select16rats of them randomly as prevention normal control group (PNC group) and treatment normal control groups (TNC group), with8rats in each group, fed with regular diet.The other120rats were taken to establish the rat model of T2DM with the method of four weeks’high-sucrose-fat diet in combination with low dose STZ (30mg/kg) injected from tail vein. The DM model rats were randomly divided into prevention model control group (PDM group), prevention TWP low-dose group (PTL group,6mg/kg), prevention TWP middle-dose group (PTM group,12mg/kg) and prevention TWP high-dose group (PTH group,24mg/kg); treatment model control group (TDM group), treatment TWP low-dose group (TTL group.6mg/kg), treatment TWP middle-dose group (TTM group,12mg/kg) and treatment TWP high-dose group (TTH group,24mg/kg), with15rats in each group. The rats of TWP prevention group began intragastric administration immediately after being into DM model and were killed after4weeks, while the TWP treatment group began intragastric administration in the5th week after being into DM model and were killed at the end of the8th week. The PNC, TNC, PDM and TDM group were treated with equal amount of distilled water. Blood glucose、24h urine total protein、blood urea nitrogen (BUN)、creatinine (Scr) were observed; kidney morphology and ultrastructural changes was observed by light microscopy and electron microscopy;The expression of the cytokine:NF-κB、 MCP-1were observed by RT-PCR and immunohistochemistry personally from the genetic and protein levels,CD68,the surface symbol protein of macrophages, was obsearved by immunohistochemistry,in order to investigate the potential mechenism of the macrophages and related cytokine in diabetic nephropathy and also the effection of the TWP Tablets.Result 1. Compared with the normal control SD rats, diabetes model of the prevention and treatment rats showed significant increased plasma glucose level and decrease in body weight, the level of KW/BW also was significantly higher in diabetes model rats(P<0.05), and KW/BW was decreased obviously with TWP treatment, and the middle dose of TWP decreased most obviously.2. The level of AST was significantly increased in PTH group compared with PNC、 PDM、PTL、PTM (P<0.05). The level of ALT、SCr、BUN、WBC、RBC、 Hb、PLT、LYM、MONO and NEUT was no significant different in prevention rats (P>0.05). Compared with the treatment normal control rats, the level of LYM was decreased markly with TWP treatment (P<0.05), and was no significant different in TTL、TTM、TTH (P>0.05). The level of AST、ALT、SCr、BUN、 WBC、RBC、Hb、PLT、MONO and NEUT was no significant different in treatment rats (P>0.05).After feeding the high-fat high-sugar diet for4weeks, Compared with the normal control rats, the level of TG、TC was significant increased in model rat (P<0.05. after TWP treatment in prevention and treatment,the level of TG、C were decrease obviously compared with model rat.3. The result of UMA. Compared with PNC and TNC group, the UMA was significantly higher in the PDM and TDM group(P>0.05); compared with the PDM group, the UMA in the TWP prevention group was reduced to varying degrees, with the most significant decline in PTM group(P<0.05), followed by PTL group (P<0.05), and the PTH group had no significant change(P>0.05);compared with the TDM group, the UMA in the TWP treatment group was significantly reduced with the most significant decline in TTM group.4. Light microscopy and electron microscopy shows: The light microscopy and electron microscopy showed that the normal control rats glomerular volume, mesangial area, the renal capsule, tubular and renal mass showed no obvious abnormalities compared with prevention and treatment groups. The glomerular and renal pathological tubular changes renal pathological lesions as DN of model control rats in the prevention and treatment groups.The pathological lesions of the group after administration of Tripterygium glycosides were improved,which group improved the most obvious is the Tripterygium wilfordii middle dose group. 5. Immunohistochemistry and PCR shows:(1) Prevention group:Compared with PNC group, CD68、NF-κB、MCP-1in PDM group was higher (P<0.05), PTL、PTM、PTH group was decreased than PDM group, and PTM decreased most obviously(P<0.05).(2) Treatment group:Compared with TNC group, CD68、NF-κB、MCP-1in TDM group was higher (P<0.05), TTL、TTM、TTH group was decreased than TDM group, and TTM decreased most obviously(P<0.05).Conclution1. The rat model of T2DM and DN were successfully prepared, and the characteristics of high blood sugar, high cholesterol and UMA were in accordance with human T2DM and DN.2. TWP Tablets can effectively delay the occurrence and development of DN before suffering from diabetic nephropathy, treatment of TWP Tablets was decreased the urine total protein lead to slow development of diabetic nephropathy, the specific mechanism is worth to research.3. The low-dose and middle-dose of TWP had better security, while the high-dose of TWP (24mg/kg) may lead to liver injury.Thus the clinical application of TWP should pay more attention to the dose and closely watched the side effects.TWP Tablets may be by restraining NF-κB、MCP-1and CD68expression to decrease the urine total protein and improve the enlarged glome rulus.4. TWP Tablets may be by restraining NF-κB、MCP-1and CD68expression to decrease the urine total protein and improve the enlarged glome rulus.5. Prevention and treatment group show the middle dose of TWP (12mg/kg/d) decreased most obviously to decrease the urine total protein and improve the enlarged glomerulus, thicked glomerular basement membrane and mesangium and was increase markedly the expressions of NF-κB、MCP-1. |
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