Effect Of Method Of Warming Kidney And Invigorating Spleen, Removing Toxicity And Activating Blood On The Serum IL-6 And TNF-α In Early-Stage Diabetic Nephropathy

ObjectiveTo investigate the effects of the method of warming kidney and invigoratingspleen, removing toxicity and activating blood (WIRA) on the inflammatoryfactor serum interleukin-6 (IL-6) and tumor necrosis factor-α(TNF-α) inearly-stage diabetic nephropathy (DN), and to explore its therapeuticmechanism.Methods1.58 cases of patients with early-stage DN with deficiency of spleen-Yangand kidney-yang, deficiency Qi and blood stasis syndrome were selected andrandomly divided into treating group(n=32) and control group(n=26). 2 groupswere given the same basal treatment including education course for diabetes,diabetic diet, and hypoglycemic and hypotension agent in routine. Patientsin the control group attended to the fosinopril(5～10mg, per os), once a day.And to those in the treating group, WIRA therapy was given additionally, onedose taken in twice a day. After 1 month's treatment, the curative effectand Chinese medical syndromes were observed. Moreover, fasting bloodglucose(FBG), post-prandial 2-hour blood glucose(P2hBG), blood lipid, urinaryalbumin excretion rate(UAE) and serum IL-6, and TNF-αwere measured andcompared before and after treatment.2. In experimental study, SPF male Sprague Dawley (SD) rats were selected,The diabetic model rats were made by a single intraperitoneal injection of60mg/kg streptozotocin(STZ). The rats were randomly assigned to four groups: normal group, model (early-stage DN)group, western medicine(WM) group withfosinopril(4mg·kg^-1·d^-1 by garage) and traditional Chinese medicine(TCM)group with WIRA complex prescription (20g·kg^-1·d^-1 by garage). The courseof treatment lasted 6weeks.The rats general status(psychosis, diet, haircolor, weight amplitude, etc), renal weight/body weight ratio, blood glucose,24hUAE, triglyceride(TG), total cholesterol(TC), serum creatinine(SCr),blood urea nitrogen(BUN). The serum IL-6, and TNF-αwere measured of ratsby enzyme linked immunosorbent assay(ELISA), and renal pathologic lesion wereobserved through light microscopy and electron microscopy. The expression oftransforming growth factor-β₁(TGF-β₁) in renal cortex of each group wereobserved by immunohistochemicl staining.Results1. Based on controlling the blood glucose availability, the total ratioof effective of TCM group was 87.5%, western medicine group is 61.5%. Comparedwith two groups, TCM group is obviously excel to western medicine group (P＜0.05). The clinical symptom score, FBG, P₂hBG and 24hUAE lowered as well asimproved the blood lipid in both groups. In TCM group, the clinical symptomscore, 24hUAE, TC, TG, LDL-C, TNF-αand IL-6 significantly reduced, the effectwas better than that in the WM group(P＜0.05 or P＜0.01). There are no toxicityand side-effect in tow groups.2. The model group rats' symptoms are energy cachexia, drinking waterincreased, urinate manifold and loose stool, lowness reaction, slow action,hair stand and lackluster. Renal slices of model group rats can be seen amajority of glomerulus increasing, swelling, expanding of capillary lumen andred blood cell depositing in it. Inflammatory cell infiltrating and mesangialwidth increasing, glomerular basement membrane(GBM) incrassation. There havenot glomerular sclerosis obviously.3. Compared with normal group, the body weight of model rats was relievedobviously(P＜0.01), Ratio of kidney weight (KW) to Body weight(BW), bloodglucose 24Huae, TC, TG, TNF-αand IL-6 significantly increased obviously(P＜0.05 or P＜0.01). the urinary volume and UAE in TCM group and WM group weredecreased, and there was better effective in TCM group. Compared with modelgroup, blood glucose, TC, TG, TNF-αand IL-6 were decreased significantlyin TCM group(P＜0.05 or P＜0.01), and there was no marked change in bloodglucose, TC, TG, TNF-αand IL-6 in WM group. 4. Compared with control group, the expression of TGF-β₁ in renal cortexof model group rats were significantly increased, espcialy in the renal tubuleand glomerular epithelium, which were all significantly inhibited by WIRAcomplex prescription. The effect of suppression over-expression of TGF-β₁ ofrenal cortex in TCM group was better than that of WM group(P＜0.05).Conclusions:1. YangQi of human body is deficient easy, and the deficiency of YangQican also incur to diabetes. Deficiency of spleen-Yang(Qi) and kidney-yang(Qi)is the onset-base of early stage DN, and internus toxicity was the importantfactor of early-stage DN. We believe that deficiency of spleen-Yang(Qi) andkidney-Yang(Qi), internus toxicity is the main pathogenesis of early-stageDN, so warming kidney and invigorating spleen, removing toxicity andactivating blood (WIRA)is the main method in curing of early-stage DN.2. The method of WIRA has better function of therapy and protection ofrelieving clinical symptom and reducing UAE of patients with early-stage DN.3. The method of WIRA has a certain effect on reducing blood glucose,regulating serum blood lipid metabolism reducing the urinary protein excretion,ameliorating pathologic change of nephridial tissue and inhibiting renalhypertrophy, mesenterium cell proliferation and thickening of GBM inexperimental diabetic rats obviously, it can also significantly inhibitedover-expression of TGF-β₁ of renal cortex.4. The method of WIRA has certain effect of anti-inflammatory onearly-stage DN, it can reduce serum TNF-αand IL-6 significantly. Thetherapeutic mechanism for early-stage DN is not depend on the controlling bloodglucose availability, but is probably pathway of relating to regulate to theproduction of inflammatory factors and anti-inflammatory reaction. It canregulate organism all round through neuro-endocrinc-immunomodulation network,and block up inflammatory factor TNF-α,IL-6 and their receptors, accordinglyregulate the expression of growth factors and cell adhesionmolecule,andinhibit multiplication of glomerular mesangium and production of ECM.