The Effect Of PCSK9Regulated By Inlammation On Renal Lipid Depostion In Mice With Adriamycin Nephropathy

PART ONE: ADRIAMYCIN NEPHROSIS MODEL ANDCHRONIC INFORMATION MODEL IN MICEObjective to copy adriamycin nephrosis model and chronicinflammation model in mice.Methods6weeks male BALB/c mice were randomly divided intocontrol group（CTL）, adriamycin group（ADR）, inflammation group（IFM）and adriamycin with inflammation group（AWI）. GivingADR andAWI one-time tail intravenous injection of adriamycin10.5mg/kg to set upadriamycin nephrosis model; CTL was given the same dose of normalsaline; IFM and AWI were given subcutaneous injection of0.5ml10%casein to replicate chronic inflammation model.Eight mice weresacrificed at4、8and12weeks after adriamycin injection.Using coomassiebrilliant blue method to detect the24h urine protein levels, automaticbiochemical analyzer to detect the serum biochemical indicators, lightmicroscope to observe morphological changes.Results ADR group and AWI group appeared a large number ofproteinuria at2weeks after injection of adriamycin, continued to12weeks(P <0.05); So did in IFM group at6weeks, but the level significantly lower than the AWI group (P <0.05). The level of blood albumin in ADRgroup and AWI group were decreased at4weeks (P <0.05); the level ofblood lipid in the two groups espectively increased in4、8weeks；the levelof serum creatinine in the two groups espectively increased in12,8weeks(P <0.05). Under light microscope, we found inflammatory cells infiltratedin renal tissue of IFM mice; ADR and AWI group appearedminimal-change nephrosis at4weeks，respectively developed into focalsegmental glomerular sclerosis with renal insufficiency at the12,8weeks.Conclusion Adriamycin nephrosis model and chronicinflammation model in mice are successful. ADR group and AWI groupearly stage are minimal-change nephrosis, late stage are focal segmentalglomerular sclerosis, but the AWI group occurred earlier than ADR group.Inflammation accelerates the development of kidney injury. PART TWO: THE STUDY OF INFLAMMATION ACTINGON PCSK9MEDIATED RENAL LIPID DEPOSTION INADRIAMYCIN INDUCED NEPHROPATHY MICEObjective To discuss the effect of inflammation acting onPCSK9-mediated renal lipid deposition in adriamycin inducednephropathy miceMethods Oil red O staining to detect fatty deposition in the kidneytissues, enzyme colorimetry to detect the contents of total cholesterol in the kidney tissues, real-time PCR to detect kidney IL-1β, TGF-β1,SREBP-2, HNF-1α, PCSK9, LDLr mRNA expression,immunohistochemical to detect kidney IL-1β, TGF-β1, SREBP-2, HNF-1α, PCSK9, LDLr protein expression.Results The content of total cholesterol in IFM group, ADR groupand AWI group were higher than control group at4weeks(P <0.05); at8weeks, total cholesterol of IFM group and ADR group were lower thanAWI group (P <0.05). IL-1β, TGF-β1, SREBP-2, LDLr mRNAexpression in IFM group, ADR group and AWI group were higher thanCTL group (P <0.05) at4,8,12weeks, the HNF-1α, PCSK9mRNAexpression were lower than CTL group (P <0.05). IL-1β, TGF-β1,SREBP-2, LDLr protein expression quantity in IFM group, ADR groupwere lower than the AWI group at8,12weeks (P <0.05), the HNF-1α,PCSK9expression quantity were higher than the AWI group(P <0.05).Conclusion The inflammation degree in mice with adriamycinnephropathy gradually increases over time. Cholesterol content inadriamycin nephrosis model with inflammation aggravate graduallyincreases, pure inflammation model with no hyperlipidemia and kidneyinjury causes a small amount of lipid deposition in kidney.Inflammationmay influence PCSK9protein by regulating the expression of SREBP-2、HNF-1α, cause renal lipid deposition.