Intervention Study Of Jiedutong Luobao Shen Capsule In Experimental Diabetic Rat Kidney Oxidative Stress And Adiponectin

Propose:Diabetic nephropathy (called DN for short) is not only the most common andmultiple clinical diabetes mellitus (called DM for short) complications, but also the mostserious microvascular complications. DN contributes to glomerulopathy of mainlymicrovascular damage. The main pathology shows it increases the permeability of capillarybasement membrane, increases the thick of basement membrane, and accumulatesglomerular mesangial extracellular matrix (called ECM for short).These changes makeglomerulosclerosis, and eventually it becomes renal fibrosis. The early clinicalperformance is glomerular hypertrophy, glomerular hyper-filterability;the following changeis micro-albuminuria, persistent proteinuria, edema, hypertension;finally it cause toprogressive development of renal insufficiency, and to renal failure at last. The etiology andpathogenesis of DN is complex and due to the multiple factors and ways. Presently themedical treatment is reducing blood sugar, reducing blood pressure, and protecting renalfunction to delay DN progression. But there is a lack of effective means of prevention andtreatment, and most patients gradually developed into renal failure and life-threatening.In recent years, the theory of oxidative stress in DN pathogenesis becomes the focustheory, which provides a theoretical basis for antioxidant therapy in the DN. Since thediscovery of adiponectin, many studies confirmed that adiponectin is closely related to DMand DN. Adiponectin is not only anti-inflammatory, anti-atherogenic effect, but also hasantioxidant properties. The reduced adiponectin levels may lead to inhibit AMPK activation,thus inhibit AMPK activating to eNOS, cause NO reducing and ROS increasing,. Thesefactors cause the aggregation of ECM in the kidney,which cause to DN. Adiponectin mayregulate the oxidative stress mechanism.Methods:This experiment about the Chinese herbal medicine jie du tong luo bao shencapsule(called JJTLBS capsule for short) of prevention DN has been studied systematically.The emphasis is to discuss the relationship between poison damage kidney networkpathogenesis theory and oxidative stress, JJTLBS capsule and adiponectin, and todemonstrate the unique advantage of JJTLBS capsule in the treatment of DN. It also confirmsthe scientific nature of the treatment of detoxification, deoppilation meridian and securitykidney, clarify the pathogenesis of DN from the toxic damage to the kidney, oxidative stress, and adiponectin.This experiment explores protection mechanism of JJTLBS capsule towardsexperimental diabetic rat kidney, and relationship between mechanism and adiponectin bystudying the kidney oxidative stress mechanisms about JJTLBS capsule interfered theexperimental diabetic rat. The experiment also provide more comprehensive theoretical andexperimental basis to prevent and treat of DN to. The experiment is divided into four parts:1The experimental type2DM rat model was made with high-fat diet combined withstreptozotocin (called STZ for short). These models divided into the DM model group, thepositive control group, the experimental group and normal control group. Close observe thegeneral state of the diabetic rats.Detect blood biochemical indices, urine albumin, insulindetermination using enzyme-linked immunosorbent assay, biochemistry and otherexperimental techniques for studying protective effects of JJTLBS capsule towardsexperimental diabetic rat kidney,2Detect fibronectin protein (FN) and type IV collagen (Col IV) by using theImmunohistochemistry assay in order to observe the ECM accumulation of JJTLBS capsuletowards experimental diabetic rat kidney. Observe the morphological changes of renal tissueby using light microscopy and transmission electron microscopy.3Detect protein expression of ADPN in serum and renal tissues, phosphorylationadenylate protein-activated protein kinase(p-AMPK),and endothelial nitric oxide enzymes(eNOS); gene expression of AdipoR1, AdipoR2and eNOS in renal tissue by using doubleantibody sandwich enzyme-linked immunosorbent assay(ELISA),immunohistochemistry,and fluorescence real-time quantitative PCR experiments to observe JJTLBS capsule towardsexperimental diabetic rat kidney.4Detect protein and gene expression of reactive oxygen species (ROS),8-hydroxy-deoxy guanosine (8-OHdG),3-nitro-casein nitrogen (3-NT) in renal tissuesby using ELISA technique to observe JJTLBS capsule towards experimental diabetic ratkidney.Results:1JJTLBS capsules can improve the general state of the experimental DM rats,decrease blood sugar, improve insulin levels and regulate dyslipidemia.2JJTLBS capsules can inhibit early renal hypertrophy and glomerular hyper-filtration state of the experimental DM rats,decrease urinary albumin excretion,which would protectrenal function.3JJTLBS capsules can inhibit the experimental DM rats morphological andultrastructural changes of rat kidney histopathology, decrease the expression of FN and ColIV in the renal tissue, decrease the ECM accumulation, which would reduce the DMglomerular sclerosis, prevent the renal fibrosis, delay the occurrence and development ofthe DN.4JJTLBS capsules can increase the serum adiponectin levels, adiponectin proteinexpression in renal tissue, AdipoR1, AdipoR2gene expression in renal tissue. While theexpression levels of AdipoR1is higher than AdipoR2.5JJTLBS capsules can increase p-AMPK protein expression in renal tissue. The geneand protein expression of eNOS were positively correlated towards adiponectin levels. Itconfirms that the increasing of adiponectin levels may enhance up-regulation the activation ofAMPK, while the AMPK active the up-regulation of eNOS.6the increased content of ROS,8-OHdG, and3-NT in the experimental DM ratssuggest that oxidative stress in kidneys play an important role in DN pathogenesis. Thecontent of ROS,8-OHdG and3-NT in kidney tissue is increased, which shows negativelycorrelated towards adiponectin levels in serum and kidney tissue. The results above showsthat the DN renal oxidative stress and adiponectin is known as the association.7JJTLBS capsules can obviously inhibit the kidney tissue content of ROS,8-OHdGand3-NT in the experimental DM rats. The pathogenesis of oxidative stress may perform likethis: it increased adiponectin levels,enhanced AMPK activation,and the activated AMPKmade the up-regulation of eNOS. Therefore, the expression of8-OHdG and3-NT werereduced and the expression of NO increased, which play a role in the protection kidneytissue.Conclusion:These results suggest that JJTLBS capsule can reduce the experimental DMrats blood glucose,increase insulin levels,regulate dyslipidemia and protect renal function,increased serum adiponectin levels, the protein and gene expression of adiponectin in renaltissue, reduce the content of ROS,8-OHdG and3-NT in renal tissue, and inhibit oxidativestress. Thus it would abatement the renal tissue damage, delay the DM development.The innovation of this experimental study is to improve the theory of detoxification, deoppilation meridian and security kidney, and introduce DN oxidative stress mechanism init. This experiment explore the relationship between DN oxidative stress in the pathogenesis,regulated by adiponectin and its receptors, and the TCM toxickidney by the theory ofpathogenesis. DN oxidative stress mechanism regulated by adiponectin and its receptors is anew idea,which may become a new target for treatment of DN,and provides new ideas andexperimental evidence when applying JJTLBS capsule for curing DN.