| Background:Diabetic Nephropathy(DN)is one of the most common and intractable complications of diabetes and it is a serious threat to human health.The etiology and pathogenesis of diabetic nephropathy are still unclear,so the study of the mechanism of DN has very important clinical value and social significance.TCM has played a huge role in the treatment of diabetes and its complications,while gaining a lot of experience.Most of the current research on the mechanisms of DN has focused on disorders of glycolipid metabolism and metabolic inflammation,cytokines and genetics,while less research has been done on endoplasmic reticulum stress and mitochondrial morphological function.Objective:Initially,the therapeutic effects of the classic Chinese medicine prescription Shenqi Wan on diabetes mellitus and diabetic nephropathy are discussed,the protective effects of Shenqi Wan on pathological damage to the kidneys are observed,and the protective mechanisms of Shenqi Wan on the kidneys are explored by analyzing the expression of endoplasmic reticulum stress and mitochondrial-related factors during the progression of diabetic nephropathy.Methods:Thirty 11-week-old C57BL/6J mice were selected as the control group,60 KK-Ay mice of the same week-old were selected as the model group and the treatment group,and the control group was fed with normal food and the model and treatment group were fed with special high-fat food.All the animals were fed and watered freely,their activity was recorded by daily observation,and their fasting blood glucose was measured and weighed every 2 weeks.10 mice of each group were killed at 16,20 and 24 weeks of age,and HbAlc and serum index tests were performed.The urine of the mice was collected for 24h 2-3 days before specimen collection and tested for urine microalbumin.Results:1.The mice in the model group showed symptoms such as lethargy,slowness of activity,eating,drinking and urinating a lot,and their body weight and fasting blood sugar were higher than those in the normal group;the mice in the treatment group showed reduced symptoms and their body weight and fasting blood sugar were lower than those in the model group.2.Urine microalbumin was significantly higher in the model group than in the normal group;it was lower in the treatment group than in the model group.3.HbAlc was significantly higher in the model group than in the normal group;it was lower in the treatment group than in the model group.4.Creatinine and triglycerides were significantly higher in the model group than in the normal group;the treatment group had a decrease compared to the model group;total cholesterol was significantly higher in the model group than in the normal group and not significantly different from the treatment group;there was no significant difference between the groups in uric acid and urea nitrogen.5.HE staining showed that the normal group had complete and clear kidney tissue structure and neat cell arrangement;the model group had thickened glomerular basement membrane,glomerular dilation and sclerosis and tubular epithelial cell edema;the treatment group had less lesions than the model group.6.PAS staining showed a small amount of glycogen staining in the glomeruli of the normal group with a pale purple coloration;the purple-red coloration in the glomerular and tubular epithelial cells of the model group was significantly deeper,suggesting the deposition of polysaccharides or glycoprotein components;the treatment group was slightly lighter than the model group.7.Masson staining showed a small amount of light blue collagen fibres in the interstitium of the normal group;blue staining of the glomeruli and interstitium of the model group,suggesting glomerulosclerosis and collagen fibre proliferation with interstitial fibrosis;the treatment group was slightly lighter than the model group.8.Immunohistochemistry showed small expression of GRP78 and OPA1 in the normal group;the model group had higher expression of GRP78 and OPA1 in the glomerular basement membrane and glomerular epithelium;the treatment group all had slightly lower expression than the model group.9.Transmission electron microscopy showed that the glomerular and tubular structures of the normal group were intact,and the mitochondria and endoplasmic reticulum were clear and intact;in the model group,the glomerular basement membrane was thickened and hardened,the cell nuclei were swollen,the podocytes were fused and disappeared,the mitochondria were fragmented,the mitochondrial cristaes were broken and disappeared,the endoplasmic reticulum was swollen and broken,and the interstitial fibrosis was less severe in the treated group than in the model group.10.Real-Time PCR measured the expression of the respective target genes for each purpose.The mRNA expression of GRP78 in the model group was significantly higher than that in the normal group,and the expression of the treated group was significantly lower than that in the model group;the mRNA expression of OPA1 in the model group was higher than that in the normal group in the early stage,but the expression level decreased significantly with the progression of the disease,and the expression in the treated group was higher than that in the model group;in the early stage the mRNA expression of MFN1 and MFN2 between the three groups was not significant,but the expression of the target genes in the model group decreased rapidly with the progression of the disease,and the expression in the treated group was significantly higher than that in the model group.11.Western Blot measured the expression of each protein for each purpose.The expression of GRP78 was significantly higher in the model group than that in the normal group and significantly lower in the treatment group than that in the model group;the expression of OPA1 in model group was higher than that in the normal group in the early stage,but the expression level decreased significantly with the progression of the disease and increased in the treatment group than that in the model group;the expression of MFN1 and MFN2 was significantly lower in the model group than that in the normal group and increased significantly in the treatment group than that in the model group.Conclusions:1.Shenqi Wan have a significant weight maintenance and fasting blood glucose reduction effect at different stages of diabetes.2.Shenqi Wan can significantly reduce the excretion rate of urinary microalbumin.3.Shenqi Wan can significantly reduce glycosylated hemoglobin levels.4.Shenqi Wan can significantly reduce the rate of creatinine excretion and have a positive effect on regulating lipid metabolism.5.Shenqi Wan are effective in ameliorating pathological changes in kidney tissues that begins at the early stages of diabetes.6.ERS occurs during diabetic kidney damage with impaired mitochondrial fusion,and Shenqi Wan plays a significant role in inhibiting ERS and protecting mitochondrial morphology. |
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