| Diabetic kidney disease(DKD)is a microvascular complication of diabetes mellitus(DM)and accounts for 40%of all cases of chronic kidney disease(CKD)worldwide.It is an important cause of end-stage renal disease(ESRD).It is characterised by a persistent increase in albuminuria excretion and/or a progressive decrease in glomerular filtration rate(GFR).Oxidative stress is the initiating cause of DKD and can contribute to lipid peroxidation,mitochondrial dysfunction,DNA damage,mediated inflammation,apoptosis and damage to glomerular structure and renal fibrosis through involvement in the regulation of glucose and lipid metabolism,leading to a sustained progressive loss of renal function.Standardised Western medical treatment still fails to halt the progression of diabetic nephropathy and there is an urgent need to explore new therapeutic approaches.Chinese medicine has anti-oxidative stress effects,and it is clinically relevant to investigate Chinese medicine to delay the progression of DKD through anti-oxidation."The formation of microscopic obstruction in the kidney luo is the pathogenesis of DKD due to the imbalance of the kidney luo.The formula "Nephrolithiasis Anti-debilitating Liquid",which is based on this theory and guided by the "Anti-Zhengjiang Method",has been proven in clinical and experimental studies.Previous basic research has shown that Nephrolithiasis Anti-decay Solution can reduce oxidative stress damage in DKD rats by increasing serum SOD levels,reducing MDA production and down-regulating renal p22phoxmRNA expression,thereby reducing inflammation and ECM accumulation,and exerting a protective effect on the kidney.However,the mechanism of anti-oxidative stress in nephritis prevention solution needs to be further investigated.In this study,the correlation between oxidative stress indicators and TCM symptoms in DKD patients was analyzed through clinical research.The pharmacological effects of Nephrolithiasis Anti-decay Solution on DKD mice and its effects on Nrf2/HO-1 oxidative stress signaling pathway were observed through animal experiments to investigate the intervention effects of Nephrolithiasis Anti-decay Solution on oxidative stress injury in DKD.Objective:1 Clinical studiesTo observe the characteristics of TCM evidence distribution and its correlation with laboratory indicators in patients with different stages of DM and DKD,focusing on the correlation between oxidative stress indicators HO-1,SOD and MDA and TCM evidence distribution.2 Animal experiments:To investigate the protective effect of nephritis prevention solution on renal injury in a high-fat feeding plus intraperitoneal injection of STZ-induced DKD mouse model and to investigate whether the mechanism is related to the Nrf2/HO-1 oxidative stress signalling pathway.Methods:1 Clinical studiesBasic information and physical and chemical findings were collected from patients with DM and DKD,and serum was retained.The patients were scored using the TCM evidence score,which was divided into two types of TCM evidence,namely,the original deficiency and the standard reality.The symptoms of Ben-Deficiency were divided into Qi-Deficiency,Yin-Deficiency and Yang-Deficiency symptoms,while the symptoms of Bid-Substantiation were divided into Qi-Depression,Damp-Heat,Phlegm-Damp and Blood-Stasis symptoms.Oxidative stress indicators are measured:serum HO-1 level,SOD level and MDA level.The laboratory physicochemical indicators,TCM evidence types and oxidative stress indicators of the patients were compared to analyse the correlation between TCM evidence types and laboratory physicochemical indicators and oxidative stress indicators.2 Animal experiments:After one week of appropriate feeding,the mice were divided into general diet and high-fat diet groups.After 8 weeks of feeding,the mice were injected intraperitoneally with STZ for 5 consecutive days to prepare the DKD model.DKD mice were considered to be successfully moulded.The mice were randomly divided into 3 groups according to their body weight,blood glucose and urinary microalbumin:10 mice in the model group,9 mice in the irbesartan group and 9 mice in the nephritis prevention solution group.Each group was given the same volume of the corresponding solution continuously for 12 weeks before sampling.Serum total cholesterol(T-CHO),triglyceride(TG)and low density lipoprotein(LDL-C)were measured in mice serum to assess lipid profile;serum alanine transaminase(ALT)to assess liver function;blood creatinine(cr),urea nitrogen(BUN),urine microalbumin and pathological tests to assess kidney damage,superoxide dismutase(SOD)and malondialdehyde(MDA)to assess The mice were tested for resistance to oxidative stress.The levels of Nrf2 and Ho-1 protein expression in mouse kidney tissues were measured by immunohistochemistry and Western Blot.To assess the effect of nephritis prevention solution on Nrf2/HO-1 signalling pathway.Results:1 Clinical studiesA total of 90 patients with DM,DKD stage Ⅲ,DKD stage Ⅳ and DKD stage Ⅴ were included in this study.The results of the analysis of laboratory findings,TCM syndrome typing and the levels of oxidative stress indicators HO-1,SOD and MDA were as follows:(1)There were statistically significant differences between the subgroups of DM,DKD stage Ⅲ,DKD stage Ⅳ and DKD stage Ⅴ in terms of duration of diabetes,duration of hypertension,presence of diabetic retinopathy,presence of history of stroke,systolic blood pressure values,serum creatinine values,cystatin C,uric acid,urea,24-hour urine protein volume,ACR,urine routine,albumin and haemoglobin.There were no significant differences between the subgroups for history of smoking,history of alcohol consumption,presence of diabetic peripheral neuropathy,presence of diabetic peripheral vascular disease,presence of diabetic foot,presence of atherosclerosis,presence of coronary artery disease,diastolic blood pressure values,blood glucose values,glycated haemoglobin,cholesterol,LDL cholesterol,HDL cholesterol and white blood cell values.(2)Oxidative stress indicators were closely related to the progression of DKD.The level of HO-1,an antioxidant stress indicator,was negatively correlated with the stage of DKD,presence of diabetic retinopathy,urea level,total 24-hour urine protein,urine albumin creatinine ratio,and urine routine grade,SOD level was negatively correlated with eGFR and urine routine grade.In contrast,MDA content,an indicator of oxidative stress,was positively correlated with disease stage,presence of diabetic retinopathy,urinary grade,total 24-hour urine protein,and urinary grade.This indicates that the damage of oxidative stress in the body becomes more and more severe as the disease progresses in DKD.(3)The types of evidence of this deficiency differed significantly between the DM group and the DKD disease groups.As the disease progressed,the proportion of Yang deficiency evidence became higher in the staging.There was no significant difference between the DM and DKD disease groups.As the disease progressed,the proportion of phlegm-damp evidence became higher in the staging and the proportion of damp-heat evidence became lower in the staging.(4)The levels of HO-1 and SOD were not significantly different among the various types of evidence of this deficiency,and the levels of MDA were lowest in Qi deficiency evidence and highest in Yang deficiency evidence,with statistically significant differences.There was no significant difference in oxidative stress indexes among the various symptoms.2 Experimental studies(1)Nephrolithiasis Anti-failure Solution can improve the symptoms of mental depression,reduced drinking and diet,retarded activity and polyuria in DKD model mice,reduce body weight,blood glucose,triglyceride(TG),total cholesterol(CHO)and low-density lipoprotein(L-DLD)levels,lower glutathione transaminase(ALT)levels,lower blood creatinine(Cr)and urea nitrogen(UAN)levels,and reduce urinary albumin excretion in DKD model mice.BUN)levels,and reduce urinary albumin excretion.(2)Nephrolithiasis Anti-failure Solution can reduce renal damage and delay renal fibrosis by increasing SOD content and reducing MDA content.The mechanism may be that Nephrolithiasis Anti-failure Solution can increase the expression of Nrf2 and up-regulate the expression of HO-1 protein,and improve oxidative stress injury in DKD mice through the Nrf2/HO-1 pathway.Conclusion1 Clinical studies:(1)Among the types of symptoms in DKD with deficiency evidence,Yang deficiency evidence is increasingly represented in the staging as the disease progresses;among the types of symptoms in DKD with solid evidence,phlegm-damp evidence is increasingly represented in the staging as the disease progresses.(2)As the disease progressed the levels of HO-1 and SOD gradually decreased between the groups and the trend of MDA was opposite to the first two,indicating that oxidative stress damage became more severe as the disease progressed.(3)The levels of HO-1 and SOD were not significantly different among the various types of evidence of this deficiency,and the levels of MDA were lowest in Qi deficiency evidence and highest in Yang deficiency evidence,with statistically significant differences.There was no significant difference in the oxidative stress index among the various symptoms.2 Animal experiments(1)Nephrolithiasis anti-debilitating solution protects the kidneys by improving metabolism and reducing urinary microalbumin excretion to reduce pathological kidney damage.(2)Nephrolithiasis Anti-failure Solution can reduce renal damage and delay renal fibrosis by increasing SOD content and reducing MDA content.The mechanism may be that Nephrolithiasis Anti-failure Solution can increase the expression of Nrf2 and up-regulate the expression of HO-1 protein,and improve oxidative stress injury in DKD mice through the Nrf2/HO-1 pathway. |
PDF Full Text Request