| BackgroundThe incidence of diabetes has increased year by year.Diabetic kidney disease(DKD)has become an important cause of end stage renal disease(ESRD),which seriously affects the health of patients and causes a heavy family and social burden.In recent years,many studies have shown that podocyte injury is closely related to DKD.The expression and distribution of podocyte-associated proteins nephrin,podocin and podocalyxin reflect the degree of podocyte injury.The research group believes that heat evil is an important pathogenesis of DKD in the early and middle stages.The clinical efficacy of DKD in the treatment of early and middle DKD is clear.On this basis,Xiaokeshenfang(Gegen,Huanglian,Shengdihuang,etc.)was developed.Based on the previous research,this study established a mouse model of early and middle DKD.By detecting the expression and distribution of podocyte-associated proteins in DKD in early and mid-term to make clear that the therapeutic effect of Xiaokeshenfang on DKD is achieved by protecting podocytesm and provide a scientific basis for the treatment of this disease.Because the etiology and pathogenesis of DKD are extremely complex,the activation of various cellular activities and signaling pathways,and the interaction of a large number of cytokines and biological activities are involved in the development of DKD.As the pathogenesis research continues,it is a single taste or compound.The advantages of multi-component and multi-target of traditional Chinese medicine provide a broad space for application.On the basis of inheriting the theory of traditional Chinese medicine,the development of a safe and effective Chinese medicine product with clear mechanism of action will benefit many patients with DKD.Mentor believes that there are "blood fever"pathogenesis in all stages of DKD,and the treatment is based on"clearing heat and cooling blood",and self-designed Xiaokeshenfang to guide the treatment to achieve a certain clinical effect.The mechanism of action of Xiaokeshenfang provides a theoretical basis for the treatment of this disease,and contributes positively to the development of modern medicine and the treatment of kidney disease.PurposeAccording to animal experiment,the effects of Xiaokeshenfang on general condition,blood glucose,24hU-Pr,renal function,renal pathology and podocyte-associated protein expression in diabetic kidney disease mice were observed,which provided experimental basis for Xiaokeshenfang in the treatment of DKD.Methods1 45 healthy male db/db mice and 15 db/m mice were selected.After 1 week of adaptive feeding(free diet and feeding),weighed and observed the basic state and randomly divided into normal groups(db/m group),model group(db/db group),western medicine group(db/db+ los.artan potassium group),Chinese medicine group(db/db+Xiaokeshenfang group),15 in each group.The Chinese medicine group was given intragastric administration of Xiaokeshenfang,the western medicine group was given intragastric administration of losartan potassium,and the normal group and the model group were given intragastric administration with same amount deionized water for 12 weeks continuously.2 The general condition of the mice was observed,and the body weight of the mice was weighed before gavage,2weeks after gavage,4weeks after gavage,8weeks after gavage,and 12weeks after gavage;and detected 24hU-Pr after 1 week of adaptive feeding and after the end of intragastric administration;blood glucose was measured 1 week after adaptive feeding and 6 weeks after intragastric administration;Detected blood renal function by abdominal aortic method after intragastric administrations;kidney samples were obtained from every group.Pathological changes of kidney samples were oberseved through paraffin section method by using PAS staining;Detected expression of podocyte-associated proteins by WB.3 All data were processed by SPSS22.0 and the difference was statistically significant at P<0.05.Results1 Mouse's General ConditionDuring the period of preparation,The mice in the normal group were generally in good condition,the weight was slowly increased,the activity was agile,the hair was shiny,the diet and the amount of water ingested were moderate;the mice in the model group were obese,lack of energy,reduced activity,weakened flexibility,rough hair,and reaction slowness;increased dietary and urine output,thin stools,skin ulcers in a small number of mice;after treatment,compared with the model group,the above conditions were improved in the Chinese medicine group and the western medicine group.Compared with the normal group,the body weight of each group of db/db mice was significantly increased(P<0.01).After treatment,compared with the model group,the body weight of the Chinese medicine treatment group and the western medicine control group decreased to different degrees(P<0.05,P<0.01);There was no significant difference between the Chinese medicine treatment group and the western medicine control group(P>0.05).2 24hU-PrBefore the treatment,the 24hU-Pr of each group of db/db mice was increased compared with the normal group(P<0.01).After intragastic administration,the 24hU-Pr of the model group?the Chinese medicine group and the western medicine control group were all higher than the normal group,the difference was statistically significant(P<0.01);Comparaed to model group,the Chinese medicine group and the western medicine control group decreased significantly(P<0.01).There was no significant difference between the Chinese medicine treatment group and the western medicine control group(P>0.05).3 Blood glucoseCompared with the normal group,the blood glucose of each group of db/db mice was significantly increased(P<0.01).After intragastic administration,compared with the model group,the blood glucose of the Chinese medicine group and the western medicine group did not change significantly(P>0.05).4 Renal functionCompared with the normal group,the urea nitrogen in the model group increased(P<0.05).Compared with the model group,the urea nitrogen in the Chinese medicine treatment group and the western medicine control group decreased(P<0.01).The serum creatinine of each group increased in different degrees compared with the normal group(P<0.05).There was no significant difference in serum creatinine between the Chinese medicine group and the western medicine group compared with the model group(P>0.05).5 Renal pathology observationThere was no obvious pathological change in the normal group.Renal pathology of the model group showed renal stenosis,thickening of the glomerular capillary basement membrane,markedly widened mesangial area,purple red glycogen,and mesangial matrix increased;renal tubule lumen Red glycogen deposition,vacuolar degeneration of local renal tubular epithelial cells,vacuolization of cytoplasm;compared with the model group,the symptom of the Chinese medicine group and the western medicine group all had alleviated in some certain.6 Expression of Nephrin,Podocin and Podocalyxin in renal cortexAfter 12 weeks of treatment,the expression levels of Nephrin protein in model group was decreased compared with the normal group.Nephrin protein of mice of the Chinese medicine group and the western medicine group were increased significantly compared with the model group(P<0.01,P<0.05).The expression of Podocin protein in model group was significantly lower than that in the normal group(P<0.01).The expression of Podocin protein in Chinese medicine group and western medicine group were increased compared with the model group(P<0.01,P<0.05).Compared with the normal group,the expression of Podocalyxin protein in the model group was decreased(P<0.05).The expression level of Podocalyxin protein in the Chinese medicine group was significantly higher than that in the model group(P<0.05).The expression level of Podocalyxin protein in the western medicine group was higher than that in the model group but there was no significant difference between the western medicine group and the model group(P>0.05).Conclusion1 Xiaokeshenfang can reduce the weight of db/db mouse.2 Xiaokeshenfang can reduce proteinuria in db/db mouse.3 Xiaokeshenfang can reduce renal pathological damage in db/db mouse.4 Xiaokeshenfang can up-regulate the expression of Nephrin,Podocin and Podocalyxin in db/db mouse podocytes. |
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