Exploring The Protective Effects Of Modified-hulubawan And The Effective Ingredient-berberine On Glomerular Podocytes In Diabetic Kidney Disease

Section 1Berberine protects against diabetic kidney disease via promoting PGC1α-regulated mitochondrial energy homeostasisObjective In this study BBR was used to treat DKD mice and cultured podocytes.The reno-protective effects of berberine were investigated,including it’s role on lipid metabolism and mitochondrial function.Methods We used blood samples from DKD patients to preform metabolomics.The db/db mice were used as DKD mouse models and cultured glomerular podocytes were used to explore the effects of BBR on mitochondrial energy metabolism.Molecular targets and mechanisms involved in the regulation of mitochondrial function and bioenergetics by berberine were investigated,along with its effects on metabolic alterations in DKD mice.Results Metabolomic analysis suggested altered mitochondrial fuel usage and generalized mitochondrial dysfunction in patients with DKD.In DKD mice,berberine treatment reversed the disordered metabolism,podocyte damage and glomerulosclerosis.Lipid accumulation,excessive generation of mitochondrial ROS,mitochondrial dysfunction,and deficient fatty acid oxidation in DKD mouse models and in cultured podocytes were suppressed by berberine.These protective effects of berberine were accompanied by activation of the peroxisome proliferator-activated receptor γ coactivator-1α(PGC-1α)signaling pathway,which promoted mitochondrial energy homeostasis and fatty acid oxidation in podocytes.Conclusions PGC-1α-mediated mitochondrial bioenergetics play a key role in lipid disorder-induced podocyte damage and development of DKD in mice.Restoration of PGC-1α activity and the energy homeostasis by berberine might be a potential therapeutic strategy against DKD.Section 2Berberine protects glomerular podocytes via inhibiting Drp1-mediated mitochondrial fission and dysfunctionObjective The mechanisms by which FFA leads to mitochondrial damage in glomerular podocytes and the effects of Berberine(BBR)on mitochondrial dynamics were explored.Methods Cultured mouse podocytes were used and treated with BBR after PA incubation.ROS levels,mitochondrial dynamics and mitochondrial function were measured to investigated the molecular mechanism of FFA-induced disturbance of and the effects of BBR.Results BBR strongly inhibited cell apoptosis,reactive oxygen species(ROS)generation,mitochondrial fragmentation and dysfunction in PA-induced mouse glomerular podocytes.Mechanistically,BBR could stabilize mitochondrial morphology in podocytes via abolishing PA-induced activation of dynamin-related protein 1(Drp1).Conclusions BBR may have a previously unrecognized role in protecting glomerulus and podocytes via positively regulating Drp1-mediated mitochondrial dynamics.It might serve as a novel therapeutic drug for the treatment of DKD.Section 3Initial research of modified-Hulubawan protects against diabetic kidney disease and its protective effects on podocytesObjective The reno-protective effects of modified-Hulubawan in DKD mouse model were investigated,including it’s role on glucose and lipid metabolism,renal function and podocyte morphology.Methods Modified-Hulubawan was used to treat DKD mice(db/db mice)for 6 weeks.Body weight,blood glucose,lipid levels,renal function,glomerular and podocyte morphology were compared between group after treatment.Molecular targets involved in the regulation of mitochondrial function were investigated.Results DKD mice treated with Modified-Hulubawan had less weight gain,lower blood glucose and reduced microalbuminuria.But the OGTT and ITT results showed modified-Hulubawan could not improve insulin sensitivity in DKD mice.Modified-Hulubawan treatment reversed the podocyte damage and glomerulosclerosis.These protective effects were accompanied by activation of the peroxisome proliferator-activated receptor γ coactivator-1α(PGC-1α)and PKM2,which promoted mitochondrial energy homeostasis and dynamics in podocytes.The HPLC identified Coptidine hydrochlorid,Pharmacoxaloid hydrochlorid,Palmatine hydrochloride,berberine hydrochloride,cinnamic acid,cinnamyl aldehyde,Fenugreek alkali andβ-ecdysterol as main ingredients of modified-Hulubawan.Conclusions Modified-Hulubawan could significantly reduce microalbuminuria,control body weight and blood glucose,inhibit podocyte damage and glomerulosclerosis.It could also promote the activity of PGC-1α and PKM2 that related to mitochondrial bioenergetics.