| Objiective:To study the effect and mechanism of Ginseng and astragalus hypoglycemic granle on ERK pathway in renal tissue of early diabetic nephropathy rats with Qi-yin deficiency syndrome,and to explore the utility and influence of ERK signal transduction pathway in the development of early diabetic nephropathy. This proves the effect of warming yang qi deficiency of medicine in the early diabetic nephropathy rats model of Qi-yin deficiency syndrome. The mechanism described Ginseng and astragalus hypoglycemic granle prevention and treatment of early diabetic nephropathy Qi-yin deficiency syndrome.It provides the basis of Chinese medicine in the treatment of early diabetic nephropathy Qi-yin deficiency syndrome.Methods:Choose SD male rats as the experimental animals,adaptive feeding for a week. random cent gives the normal control group and diabetic nephropathy module, diabetic nephropathy module to build model rats left nephrectomy, conventional anti-inflammatory five dayshigh sugar and high fat feed for 4 weeks after 1 week.To eliminate the rats which did not meet the requirements of the experiment, one-time intraperitoneal injection of urea with little dose of chain rhzomorph replication model of diabetic nephropathy, Building requirement rats were randomly divided into diabetic nephropathy model(model group), qi-yin-Deficiency treatment group,qi-yin-Deficiency treatment group. the normal control group and model groupgave little quantity normal saline,qi-yin-Deficiency treatment group、qi-yin-Deficiency treatment group all gaved Chinese medicine elixation of Ginger、Cinnamon、Green Husk. Meanwhile qi-yin-Deficiency treatment group gaved declining sugar granule,four groups gaved corresponding intervention.Intervention in 8 weeks. each group rats were executed. after testing each rat urine trace albumin excretion rate(UAER) and urine creatinine(available),serum creatinine,fasting glucose, glycosylated hemoglobin(GHb), 24 h urine was measured,ELISA method to detect fiber connections in the serum protein(FN) content; and calculate the endogenous creatinine clearance rate(CCr), kidney weight/body weight;HE dyeing method to observe each kidney pathological form;Immunohistochemical method to detect groups of ERK protein involved in kidney tissue ERK signal transduction road, the expression of MKP 1and p– ERK.Results:1. In the aspect of blood sugar levels: compared with normal group, the model group(p<0.01)and the qi-yin-Deficiency group(p<0.05)were significantly higher, there was no statistically significant difference between qi-yin-Deficiency treatment group and normal group;Compared with model group,qi-yin-Deficiency group and qi-yin-Deficiency treatment group both significantly lower(p<0.01),found statistically significantly;there were no statistically significant difference between qi-yin-Deficiency group and qi-yin-Deficiency treatment group.2. In the aspect of blood serum FN: compared with normal group, the model group, the qi-yin-Deficiency group(p<0.05)were significantly higher,found statistically significantly, there was no statistically significant difference between qi-yin-Deficiency treatment group and normal group;Compared with model group,other groups no statistically significant difference;there were no statistically significant difference between qi-yin-Deficiency group and qi-yin-Deficiency treatment group.3. In the aspect of blood GHb: compared with normal group, other three groups were higher,found statistically significantly(p<0.01);Compared with model group,qi-yin-Deficiency group and qi-yin-Deficiency treatment group were lower, found statistically significantly(p<0.01);There were statistically significant difference between qi-yin-Deficiency group and qi-yin-Deficiency treatment group(p<0.01).4. In the aspect of kidney index: compared with normal group,Other three groups were higher,found statistically significantly(p<0.01);Compared with model group,qi-yin-Deficiency group(p<0.05)and qi-yin-Deficiency treatment group(p<0.01) were lower, found statistically significantly;There were statistically significant difference between qi-yin-Deficiency group and qi-yin-Deficiency treatment group(p<0.05).5. In the aspect of urine volume ml/24h: compared with normal group,other three groups were higher,found statistically significantly(p<0.01);compared with model group,qi-yin-Deficiency group and qi-yin-Deficiency treatment group were lower(p<0.01),found statistically significantly;There were no statistically significant difference between qi-yin-Deficiency group and qi-yin-Deficiency treatment group(p>0.05).6. In the aspect of blood UAER: Compared with normal group, other three groups were higher,found statistically significantly(p<0.01); compared with model group, other three groups were difference,found statistically significantly(p<0.01);There were no statistically significant difference between qi-yin-Deficiency group and qi-yin-Deficiency treatment group(p>0.05).7. In the aspect of blood CCr: compared with normal group,model group and qi-yin-Deficiency group were lower,found statistically significantly(p<0.01),qi-yin-Deficiency treatment group no difference,no statistically significantly; compared with model group,qi-yin-Deficiency group were lower,found statistically significantly(p<0.05);There were no statistically significant difference between qi-yin-Deficiency group and qi-yin-Deficiency treatment group(p>0.05).8. In the aspect of urinary creatinine values: compared with normal group, model group and qi-yin-Deficiency group were lower,found statistically significantly(p<0.05),qi-yin-Deficiency treatment group no difference,no statistically significantly(p>0.05);compared with model group,qi-yin-Deficiency treatment group had no significant difference,There were statistically significant difference between qi-yin-Deficiency group and qi-yin-Deficiency treatment group(p<0.01).9. There were no statistically significant difference both two comparision between groups.10. Renal tissue HE staining of rats in each group,examined under a microscope(500-times),pathological imaged under light microscope pathology picture:normal renal tissue of normal rats; model group rats of glomerular part slight hyperplasia of mesangial matrixed, part of the glomerulus see table with scattered in mesangial cell proliferation; the rat kidney pathological changes of qi-yin-Deficiency group were compared with model group serious, focal hyperplasia of glomerular mesangial matrix, many parts see glomerular segmental hyperplasia;qi-yin-Deficiency treatment group were compared with model group light, only focal hyperplasia of glomerular mesangial matrix, some see glomerular segmental hyperplasia;compared with glomerular mesangial proliferation change has no obvious difference.each rat kidney glomerular sclerosis was found.11. Immunohistochemical detection of nomal group rats no found the expression of ERK、MKP-1 and p-ERK, the rest of the group rats glomerulus and renal tubule cells were expressed,see scattered yellow spotting. Compared with model group,the expression of yellow spotting qi-yin-Deficiency group increase, the expression of yellow spotting qi-yin-Deficiency treatment group reduce.12.ERK protein expression in the average optical density value aspects: compared with normal group, model group and Yin deficiency of higher value(p<0.01), higher values of gas Yin deficiency of the treatment group(p<0.05); Compared with model group, qi and Yin deficiency treatment group obviously low value(p<0.01), qi and Yin deficiency two set of values, no significant difference; Compared with qi and Yin two virtual group, qi and Yin deficiency treatment group value reduced significantly, the difference was statistically significant(p<0.01).13.Protein expression in MKP-1 the average optical density value aspects: compared with normal group, model group and Yin deficiency of group increased significantly(p<0.01), qi and Yin deficiency treatment group value higher(p<0.05); Compared with model group, qi and Yin deficiency group two values on the high side(p<0.01); Qi and Yin deficiency treatment group value lower(p<0.01); Compared with qi and Yin two virtual group, qi and Yin deficiency treatment group value higher(p<0.01).14.In terms of average optical density value of p-ERK expression: compared with normal group, model group and Yin deficiency of group increased significantly(p<0.01), qi and Yin deficiency treatment group has no obvious difference; Compared with model group,low values of gas Yin deficiency of the treatment group(p<0.01), qi and Yin deficiency two set of values increased(p<0.01); Compared with group gas Yin deficiency of qi and Yin two virtual treatment group values on the high side(p<0.01).Conclusion:Chinese medicine ginger,cinnamon,blue aggravate the rats with qi and Qi-yin deficiency syndrome,witch has a protective effect on biochemical markers of also have a significant effect to the kidneys of rats.Ginseng and astragalus hypoglycemic granle can reduce blood glucose,GHb,serum FN,reduce urinary protein,inhibit the renal tissue ERK signaling pathway protein ERK,p- ERK and MKP-1 expression,witch can inhibit mesangial proliferation,prevent glomerular fibrosis and protect renel function,delay the progression of diabetic nephropathy with Qi-yin deficiency syndrome. |
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