The Changes Of Drug Metabolic Enzymes And Transporters In Nephrotic Syndrome And The Effect Of Chinese Medicine On Its Hyperlipidemia

Background and ObjectivesNephrotic syndrome(NS)is a common metabolic disease with systemic immune and inflammatory disorders.Glucocorticoid is the preferred drug for clinic treatment and a combination of drugs is often used because of its various complications.As one of its important manifestations,hyperlipidemia can not only increase the cardiovascular susceptibility of patients,but also cause a lipid deposition in the kidney,aggravate the inflammation and promote the development of glomerulosclerosis.In order to control the blood lipid,statins are often applied in combination with hormones.The drug combination is more likely to increase the occurrence of adverse reactions and makes the prognosis difficult.There has no report on the dynamic changes of the expression of metabolic enzymes and transporters in the disease state.Both Rhizoma Imperatae and Hedyotis diffusa have good anti-inflammatory and immunomodulatory activities.It’s reported the polysaccharides of Rhizoma Imperatae can improve glucose and lipid metabolism.They are commonly used in the treatment of acute and chronic glomerulonephritis,and could be the candidate drugs for the treatment of NS.However,the mechanism of their lipid-lowering effect on NS has not been reported.This paper aimed to study the following aspects:(1)the dynamic changes of metabolizing enzymes and transporters in NS;(2)the intervention effect of Rhizoma Imperatae and Hedyotis diffusa on dyslipidemia in NS.Methods1.The rat model was established by a single injection of Adriamycin(6 mg/kg).The expression changes of major metabolizing enzymes and transporters in the liver and kidney of AN rats were detected from 6th,9th and 12th week after the injection.2.LC-MS/MS method was used to identify the components of the decoction.After the successful establishment of AN model,AN rats were orally administrated decoction for 4 weeks.The renal function index,blood lipid index and inflammatory factors were detected to show the therapeutic effect of decoction on AN rats.PCR and Western blot were used to study the intervention mechanism of decoction on dyslipidemia in AN rats.The pharmacodynamic material basis of the decoction was predicted by molecular docking technique.Results1.In AN model,the expression of major metabolizing enzymes and transporters in the liver and kidney changed with the development of NS.In liver,the expression of Oatp1b2,Oct 1,Ntcp,Mdrl,Mrp2,Bsep and Mrp4 mRNA decreased in different periods and the mRNA expression of Bcrp increased.The protein expression of Octl and Mdrl was consistent with the changes of their mRNA expression.Besides,the mRNA expression of the main metabolic enzymes(Cyp3a4,2d6,2a9,7a1,7b1,27a1,etc.)decreased.In kidney,the mRNA expression of Mrp2 and Mrp4 increased,while the expression of Oatl kept decreased.Oat2 and Oct2 mRNA showed a decreased expression at 6th and 9th week and an increased level at 12th week.The mRNA expression of Mdr1 in AN group was higher than that in normal group at 6th week.The protein expression of Oct2 and Mdrl was in accordance with the mRNA trend.2.The decocotion of Rhizoma Imperatae and Hedyotis diffusa contained asperulosideacid,scandoside,desacetyl asperulosidic acid,chlorogenic acid and so on.3.The symptoms of AN rats treated with decoction were improved as follows:(1)The level of urine protein(upro)and uric acid(UA)decreased and the renal function was improved;(2)The plasma level of cholesterol(TC),triglyceride(TG)and low-density lipoprotein(LDL)decreased,suggesting an improvement of blood lipids;(3)The expression of tumor necrosis factor α(TNF-α)and interleukin 1β(IL-1β)was down-regulated,showing the inflammatory response was inhibited;(4)In liver,AN rats had a decreased mRNA level of Cyp7bl and PPARy,which was raised after decoction treatment.Besides,AN rats showed an increased expression of PCSK9 and a decreased level of LDLR.Whereas,the protein expression of PCSK9 was decreased and LDLR expression was increased after treatment;(5)A higher expression of PCSK9 and lower level of LDLR were observed in the kidney of AN rats treated with the decoction.The decoction also exerted an alleviation effect on the renal inflammatory reponse to reduce the level of IL-1β,TNF-α,MCP-1 and TGF-β1.These results suggested that the lowering lipid mechanism of decoction on AN rats may work through PPARγ-CYP7B1 and PCSK9-LDLR signaling pathway and kidney inflammation was inhibited at the same time.Besides,it was found that flavonoids and iridoids had the highest binding force with targets,which may be the main effective components in the decoction.