Clinical And Basic Research Of Shenkang Preparation Alleviating Protienuria And Podocyte EMT In Diabetic Kidney Disease By Improving Insulin Resistance

[Background/Aims]In our country,diabetic kidney disease(DKD)is an important cause,resulting in the end-stage renal disease(uremia)in diabetes(DM)patients.For the DKD patients with tertiary prevention,"educing proteinuria" should be a conventional therapeutic measure in the integrated treatment of DKD,but currently,there is a lack of the effective drugs in clinic.Studies have shown that insulin resistance(IR)is the upstream pathogenic factor of proteinuria in DKD patients;Combined with the basic conventional treatments,Shenkang injection(SKI),a preparation of Shenkang,or Levocarnitine injection(LI)can reduce urine protein in some DKD patients,respectvely.Therefore,the authors supposed that,for the early DKD patients,combined with the basic conventional treatments,SKI or LI can not only improve IR,but also reduce proteinuria,and then delay the progression of DKD.[Methods]A total of the 90 DKD patients with G2 and A2 stage were collected.The patients were equally divided into 3 groups,the A group,the B group and the C group using a random number table(the 7 cases were lost at the end of the trial).Prior to the trial,the baselines of patients including age,gender,duration of illness,body mass index,blood pressure,whether or not combining hypertension,antihypertensive drugs(the use of ACEI/ARB in hypertensive patients)and oral hypoglycaemic agents(the use of insulin secretagogues/non-insulin secretagogues)were compared among the 3 groups.All the patients in the 3 groups received the basic conventional treatments.Based on these conventional treatments,the patients in B group were added daily SKI by intravenous infusion,while the patients in C group were added daily LI by intravenous infusion.After the intervention of drugs for 2 weeks,the general indicators including gender,age,duration of illness,body mass index(BMI),blood pressure(BP),whether or not combining hypertension,the use of ACEI/ARB in hypertensive patients,the use of insulin secretagogues/non-insulin secretagogues,tongue appearance and pulse condition,the scores of traditional Chinese medicine(TCM)syndromes,IR indicators including fasting blood glucose(FBG),fasting insulin(FIN),homeostatic model assessment of insulin resistance(HOMA-IR),urinary protein indicators including urine microalbumin(micro-UAlb),urine microalbumin/urine creatinine(UACR)and other indicators were investigated respectvely in the 3 groups.[Results]The clinical characteristics of TCM in the 83 DKD patients with G2 and A2 stage were basically in line with the syndrome criteria of "syndrome of spleen and kidney qi deficiency,blood stasis and turbidity and poison";SKI could improve TCM syndromes,IR indicators such as FIN and HOMA-IR,while LI had no effects on IR;Both SKI and LI could reduce micro-UAlb and UACR,moreover,the improved effects of SKI were better than LI;Neither SKI nor LI had the effects on renal dysfunction,blood lipids,glycated hemoglobin,hepatic dysfunction,red blood cell counts and hemoglobin.[Conclusions]For the 83 DKD patients with G2 and A2 stage,SKI can ameliorate TCM syndromes,IR indicators and microalbuminuria,and that the improved effects of SKI were better than LI.[Background/Aims]Insulin resistance(IR)and its related signaling pathways activity disorder in the kidney are the upstream pathogenic factors,which induce podocyte epithelial-mesenchymal transdifferentiation(EMT),further resulting in podocyte injury in diabetic kidney disease(DKD).IR-related signaling pathways,except the classical IRS1/PI3K/Akt pathway,also include Erk1/2 pathway,GLUT4 pathway and TGF-β1/Smads pathway.In clinic,the efficacy of Shenkang injection(SKI)treating proteinuria in DKD patients has been widely recognized,however,its effects and mechanisms are still unclear.Therefore,using the modified DKD rat model,the authors tried to elucidate the effects and mechanisms in vivo of Shenkang granules(SKG),a preparation of Shenkang,inhibiting podocyte EMT by regulating IR and its signaling pathways.[Methods]The modified DKD rat model was established by the combinative 3 measures including high fat feed,unilateral nephrectomy and streptozotocin(STZ)intraperitoneal injection.The experimental rats were divided into 4 groups,in which,except the normal group,the DKD model rats were further randomly divided into the model group,the SKG-treated(SKG)group and the rosiglitazone-treated(ROS)group.The rats in the normal group and the model group were daily oral administrated with double distilled water(2 mL),while the rats in the SKG group and the ROS group were daily oral administrated with SKG suspension(3 g kg-1and ROS suspension(5 mg kg-1)respectively.After the administration of drugs for 6 weeks,all rats were sacrificed.Their urine,blood and renal tissues were collected to examine the various indicators,and then,investigating the effects of SKG and ROS on the IR-related indicators,the IR signaling pathways-related indicators,the podocyte EMT indicators,the glomerular lesions-related indicators and the efficacy indexes in the model rats.[Results]For the modified DKD model rats,(1)There were IR in the DKD model rats;Both SKG and ROS could improve IR,but the effects of SKG were less than ROS.(2)The IR-related signaling pathways in podocyte of the DKD model rats included TGF-β1/Smads pathway,GLUT4 pathway and Erkl/2 pathway;The intervention mechanisms of SKG might be regulating the protein expressions of the key signaling molecules in the above-mentioned signaling pathways,and that the targets of SKG were similar to ROS.(3)In the DKD model rats,the IR-induced features of podocyte EMT were the changes of the expression extent and the protein expression level in nephrin,P-Cadherin,Collagen-1 and desmin in the kidney;The intervention effects of SKG were to improve the expression extent and the protein expression level in nephrin,P-Cadherin,Collagen-1 and desmin in the kidney,moreover,the effects of SKG were similar to ROS.(4)In the DKD model rats,the pathological features of the podocyte EMT-related glomerular lesions included glomerulosclerosis,GBM thickening and foot process form changes;The intervention effects of SKG were to improve glomerulosclerosis,GBM thickening and foot process form changes,moreover,the effects of SKG on glomerulosclerosis and GBM thickening were better than ROS.[Conclusions]The effects in vivo of SKG inhibiting podocyte EMT in the DKD model rats were achieved by regulating IR and its signaling pathways activity.