Clinical Study On The Effect Of Qiwei Granule On Diabetic Nephropathy And Its Protective Mechanism Of Podocytes

Objective:The aim of our clinical study was to evaluate the effect and safety of Qiwei granule onDiabetic nephropathy (DN) patients inclass Ⅱ or class III with deficiency of Qi and Yin and blood stasis in channels. In the animal study,our aim was to investigate the effect of Qiwei granule on the podocyte in KK-Ay mice renal tissue and underlying mechanism.Methods:In according with the principle of random, control, and double blind and the technical requirements of new traditional Chinese medicinein clinical study, we recruited48DN patientsin class II or class III for our observation. The patients were allcharacterized with syndrome of deficiency of Qi and Yin and blood stasis in channels from Guang’anmenHospital36cases of them were treated with Qiwei granule and the other12cases were treated with placebo for12weeks.During the treatment, we recorded the vital signs, blood glucose, glycosylated hemoglobin, blood routine, urine routine, feces routine, liver function (including AST, ALT, ALB) and renal function (including Cr, BUN) as the safety indexes, and monitored adverse reaction.In addition, the24h microalbuminuria and24h proteinuria of all the patients were examined, and the traditional Chinese medicine syndrome were observed in our study. In addition,35male KK-Ay mice after acclimating to new surroundings for4weeks, were randomly divided in model group, Irbesartangroup, low-dosage Qiwei granule group, middle-dosage Qiwei granule groupand high-dosage Qiwei granule group, while8C57BL/6J mice were used as normal control. Body weight, blood glucose levels and24h proteinuria were measured. After10weeks treatment, we measured the biochemistry parameter and kidney weight, and histological analysis including hematoxylin and eosin (HE), Masson’s Trichrome, periodic acid-Schiff (PAS), transmission electron microscopy and immunohistochemical staining for WT-1. The protein expression of nephrin, JNK, pJNK was detected by Western Blot and mRNA expression of nephrin, a3integrin,and βlintegrin was measured by RT-PCR.Results:After12week treatment, Qiwei granule obviously alleviated the symptoms of DN patientswith syndrome of deficiency of Qi and Yin and blood stasis in channels. Except3shedding case in Qiwei granule group,33cases were significantly changed and30cases were effective in the traditional Chinese medicine syndrome. Qiwei granule significantly decreased the24h microalbuminuria and24h proteinuria. In the33cases treated with Qiwei granule,14 cases were clinical-controlled and3cases were significantly changed in the renal function effective evaluation, while the total effective rate was52%. But there was no significant difference in blood pressure, blood glucose, glycosylated hemoglobin, creatinine clearance between Qiwei granule group and placebo group which were all in reasonable level. Compared with placebo group, the safety indexes of Qiwei granule group such as blood routine, urine routine, feces routine, liver function (including AST, ALT, ALB) and renal function (including Cr, BUN) did not alter before or after the treatment.In animal study, the body weight, blood glucose, total cholesterol, total triglyceride, kidney weight, and serum creatinine levels of the model group were significantly increased compared with the normal group after10weeks treatment. The three different dosage Qiwei granule groups and Irbesartangroup all decreased the body weight, blood glucose,24h albuminuria, and serum creatinine. Thekidney weight inthree different dosage Qiwei granule groups tended to be lower than that in model group, although the difference was not statistically significant. We observed the glomerular fibrosis, glomerular mesangial matrix expansion, glomerular basement membrane expansion, capillary thickness, renal interstitial fibrosis with chronic inflammatory cell infiltrating, and kimmelstiel-wilson nodule in model group. While the three different dosage Qiwei granule groups and Irbesartan group ameliorated these renal lesions. There were fusions of foot processes of podocyte, podocyte detachment in model group under transmission electron microscope, while the treatment groups ameliorated the podocyte injury. The podocyte number marked with WT-1in model group was decreased, while the treatment groupspreserved it. In addition, the protein and mRNA expression of nephrin, the α3integrin and βlintegrin mRNA expression was significant increased in the three different dosageQiwei granule groups and Irbesartangroupcompared with model group. The protein expression of pJNK, pJNK/JNK of the three different dosage Qiwei granule groups and Irbesartan group were lower than model group.Conclusion:It has been proved that Qiwei granule is effective and safety on Diabetic nephropathy (DN) patients inclass Ⅱ or class Ⅲ with deficiency of Qi and Yin and blood stasis in channels.Qiwei granule alleviates the glucose and lipid metabolism in KK-Ay mice, mitigates the renal lesion, and protects podocyte to prevent the progression of DN.Qiwei granule attenuates the podocyte injury in the diabetic nephropathy ofKK-Ay miceby upregulating the expression of nephrin, a3integrin and βlintegrin and inhibiting JNK signal pathway.