| Sanhuang Yishen Capsule(preparation of quasi Name: Z20050795) is a preparation in hospital Cangzhou integrated traditional Chinese and Western medicine for early diabetic nephropathy, clinical, early studies have been carried out on the part of mechanism, this topic is on the basis of previous studies, an in-depth study on Mechanism of action.Objective: To observe the Sanhuang Yishen Capsule on kidney of diabetic rat matrix metalloproteinase-9(MMP-9) and tumor necrosis factor alpha(TNF-α) influence, so as to understand the mechanism of this based on the clinical application of Sanhuang Yishen capsules in the treatment of diabetic nephropathy and provide a theoretical basis.Method: 60 clean adult male Wistar rats were randomly divided into two groups, 10 rats in the normal group, model group with 50 rats. The model group rats were fasted 12 h after intraperitoneal injection of streptozotocin(STZ) 60mg/kg(STZ before clinical use of 0.1M sodium citrate buffer solution with the concentration of 1%, p H = 4.2 solution), sodium citrate buffer was injected normal group. Blood collected from the tail vein after 72 h,blood glucose value is more than 16.7mmol / L, identified as the diabetic rats.After the success of Modeling Rats 40 were randomly selected and divided into model group(10 rats) and Sanhuang Yishen Capsule low dose treatment groups(10 rats), dose of Sanhuang Yishen Capsule treatment group(10 rats),Sanhuang Yishen Capsule high dose treatment group(10 rats). A week after modeling, traditional Chinese medicine treatment group rats dose according to the human body surface area ratios of equivalent dose conversion in rats.Sanhuang Yishen Capsule low dose treatment group were given 0.4g/(kg- D)by gavage, Sanhuang Yishen Capsule medium dose treatment group weregiven 0.8g/(kg- D) by gavage, Sanhuang Yishen Capsule high dose treatment group were given 1.6g/(kg- D) by gavage daily by gavage, 1 time, 8 weeks in a row. The normal group and model group were given the same dose of saline daily gavage. During the experiment, rats fed a standard diet, free drinking water, the environment temperature is 22 DEG to 26 DEG C, humidity45%-65%, insulin and other antidiabetic drugs not used during the experiment.8 weeks after administration, the tail vein blood glucose. Intraperitoneal injection of anesthetized rats, abdominal incision, abdominal aortic blood serum, centrifugal separation. Detection of serum triglyceride automatic biochemical analyzer(TG), cholesterol(TC), creatinine(SCr), blood urea nitrogen(BUN), serum C reactive protein(CRP). After took blood from the rats, both kidneys removed, removal of the renal capsule and renal hilar connective tissue with physiological saline wash after weighing, 10% neutral formalin solution in fixing, embedding, slicing, MMP-9 and TNF-alpha detected by immunohistochemistry. Kidney tissues were fixed in 10%formalin, dehydrated, embedded in paraffin, slice thickness 4 μm, hematoxylin eosin(hematoxylin-eosin staining HE) staining to observe the pathological changes of glomerulus and renal tubule interstitial under microscope.Application of Statistics Package for Social Sciences 16.0(SPSS16.0) analysis software for statistical processing. The measurement data is represented by?x±s group compared with single factor analysis of variance, group before and after treatment were compared by paired t test. Count data using X2 test.Results:1 Effects of ratGeneral state of the diabetic rat group mental state is poor, the amount of water increased, urine volume increased, the hair color dull dull.2 weight changes of the ratsIn weight compared with no difference(P>0.05); before the treatment,compared with the normal group, model group and Sanhuang Yishen Capsule low, middle, high dose treatment groups without significant changes in body weight(P>0.05), no significant differences were found in the model group andSanhuang Yishen capsule low, high dose treatment groups(P>0.05); the body weight of rats by intragastric administration after 8 weeks of treatment,compared with the normal group, model group and Sanhuang Yishen Capsule low, middle, high dose treatment groups significantly decreased body weight(P<0.05), model group and Sanhuang Yishen Capsule low, high dose treatment group with no significant difference between the body weight(P>0.05); rats of normal group after treatment than before treatment significantly increased body weight(P<0.05), model group and Sanhuang Yishen capsules, in low, high dose treatment groups of rats after the treatment of body weight were significantly lower than those before treatment(P<0.05).3 kidney weight / weight ratio influence by intragastric administrationAfter gavage after 8 weeks of treatment, compared with the normal group,model group and Sanhuang Yishen Capsule low, middle, high dose treatment group, kidney weight / body weight ratio was significantly increased(P<0.05);compared with the model group, low dose of Sanhuang Yishen Capsule treatment group, kidney weight / body weight ratio had no significant difference(P>0.05) in the treatment group, high dosage; kidney weight / body weight ratio was significantly reduced(P<0.05); compared with Sanhuang Yishen low dose group, high dose treatment group, kidney weight / body weight decreased significantly(P<0.05); high dose treatment groups, ratio of kidney weight / body weight had no significant difference(P>0.05).4 Effects of biochemical indexes4.1 Effect of glucose treatment compared with the normal group, blood glucose before diabetes rats, were significantly increased(P < 0.05), but no significant difference in blood glucose of diabetes among the groups(P >0.05), comparable. After gavage after 8 weeks of treatment, there was no significant difference compared with the change of blood glucose levels before and after treatment(P> 0.05); the model group and Sanhuang Yishen Capsule low, middle, high dose treatment groups had no significant difference between group comparison of variations(P> 0.05), there was significant difference compared with the normal group(P < 0.05).4.2 After 8 weeks the effect of serum cholesterol, glycerin three greases,compared with the normal group, model group, treatment group, three rats serum glycerin fat, cholesterol level increased significantly(P < 0.05).Sanhuang Yishen Capsule low, medium and high dose treatment group compared with the model group, no significant difference of glycerin three fat,cholesterol levels(P > 0.05). Model group, Sanhuang Yishen Capsule low,medium and high dose treatment groups compared the cholesterol, glycerin three greases have no significant difference(P > 0.05). Comparison of Sanhuang Yishen Capsule low dose treatment groups and Sanhuang Yishen Capsule in middle and high dose treatment groups, no significant difference of glycerin three fat, cholesterol levels(P> 0.05). Sanhuang Yishen Capsule, high dose treatment group comparison, no significant difference of glycerin with three fat, cholesterol levels(P> 0.05).4.3 After 8 weeks the effect of serum creatinine, urea nitrogen, compared with the normal group, the model group rats serum creatinine, urea nitrogen level increased significantly(P< 0.05); Sanhuang Yishen Capsule low dose treatment group, serum creatinine, urea nitrogen level increased significantly(P < 0.05); Sanhuang Yishen Capsule, high dose treatment groups had no significant difference in serum creatinine, urea nitrogen levels(P > 0.05).Compared with the model group, Sanhuang Yishen Capsule low dose treatment groups had no significant difference in serum creatinine, urea nitrogen levels(P> 0.05); Sanhuang Yishen Capsule in the treatment group,high dose of serum creatinine, urea nitrogen levels were significantly decreased(P < 0.05). Compared with Sanhuang Yishen Capsule low dose treatment groups, Sanhuang Yishen Capsule in treatment group, high dose of serum creatinine, urea nitrogen levels had significant difference(P< 0.05).Sanhuang Yishen Capsule, high dose treatment group comparison, no significant difference of serum creatinine, urea nitrogen levels(P> 0.05).4.4 8 weeks after the effects of ultra sensitive C response protein in serum,compared with the normal group, the serum level of C reactive protein in rats of model group increased significantly(P< 0.05); Sanhuang Yishen Capsulelow dose treatment group, serum C reactive protein level increased significantly(P < 0.05); no significant difference of Sanhuang Yishen capsules high dose treatment group, serum level of C reactive protein(P > 0.05).Compared with model group, no significant difference of Sanhuang Yishen Capsule low dose treatment group, serum level of C reactive protein(P >0.05); Sanhuang Yishen Capsules in middle and high dose treatment markedly decreased the level of serum C-reactive protein in group C(P < 0.05).Compared with Sanhuang Yishen Capsules low dose treatment groups,Sanhuang Yishen Capsule in middle and high dose treatment markedly decreased the level of serum C-reactive protein in group C(P < 0.05).Sanhuang Yishen Capsules in middle and high dose treatment group comparison, no significant difference of serum C reactive protein levels(P >0.05).5 Effects of MMP-9 and TNF- alphaThe normal expression of MMP-9 in normal rat kidney, compared with the normal group, the model group rats, expression decreased obviously(P < 0.05);Sanhuang Yishen Capsule low dose treatment groups expression decreased obviously(P < 0.05); Sanhuang Yishen capsules in the treatment group, high dose of expression has no significant difference(P > 0.05). Compared with the model group rats, Sanhuang Yishen Capsule low dose treatment groups no significant difference(P> 0.05); high dose treatment groups, the expression increased significantly(P< 0.05). Compared with Sanhuang Yishen Capsule low dose group, middle and high dose treatment group expression was significantly increased(P< 0.05). Sanhuang Yishen capsules in middle and high dose treatment group comparison, expression has no significant difference(P> 0.05). A small amount of TNF- alpha expression in normal rats,compared with the normal group, the model group rats, the expression increased significantly(P< 0.05); Sanhuang Yishen Capsule low dose treatment groups significantly increased the expression(P < 0.05); Sanhuang Yishen capsules in the treatment group, high dose of expression has no significant difference(P> 0.05). Compared with the model group, low dose ofSanhuang Yishen Capsule treatment group no significant difference(P > 0.05);Sanhuang Yishen capsules in the treatment group, high dose of expression decreased obviously(P < 0.05). Compared with Sanhuang Yishen Capsule low dose treatment groups, Sanhuang Yishen Capsule in middle and high dose therapy expression decreased obviously(P < 0.05). Sanhuang Yishen capsules in middle and high dose treatment groups had no significant difference(P >0.05).6 Observation of light microscope showed renal histopathologyNormal control group rat kidney glomerular and renal tubular structure was normal, renal interstitial inflammatory cell infiltration, vascular dilatation and congestion free; model group glomerular endothelial cell swelling,mesangial cells increased; renal tubular epithelial cells, and even necrosis;renal interstitial inflammatory cell infiltration, vascular dilation and hyperemia.Compared with the normal group, low dose of Sanhuang Yishen Capsule treatment group rat kidney glomerulus and renal tubule structure change significantly, middle and high dose treatment group was close to the structure change. Compared with the model group, low dose of Sanhuang Yishen Capsule treatment group had no obvious change; the middle and high dose treatment groups of glomerulus and renal tubule lesion alleviated the structure.Compared with the three yellow Yishen Capsule low dose group, middle and high dose treatment group of glomerulus and renal tubules lesion alleviated the structure. But in high dose group, with no significant difference between the.Conclusion:1 Glucose and lipid metabolism in Sanhuang Yishen Capsule on disorder of no treatment effect.2 Shanhuang Yishen Capsule could reduce the blood urea nitrogen,creatinine, serum C reactive protein level.3 Shanhuang Yishen Capsule can inhibit the expression of MMP-9 in kidney decline, inhibition of TNF-alpha expression in kidney, relieve diabetic rat kidney hypertrophy.4 This experiment showed that three of the patients with renal capsule and high dose had a therapeutic effect on diabetic nephropathy, but there was no significant difference between the two groups.5 copy of this experimental model of diabetic rats after 8 weeks, blood urea nitrogen, creatinine, serum CRP increased significantly, kidney MMP-9expression decreased, the expression of TNF-alpha increased, but Shanhuang Yishen Capsule after 8 weeks of treatment, blood urea nitrogen, creatinine decreased significantly, reduce renal damage, significantly decreased serum CRP, MMP-9 expression in kidneys increased, the expression of TNF-alpha decreased, which indicated that the damage of kidney Shanhuang Yishen Capsule through the above mechanisms that delay in diabetic rats. |
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