Expression Of NF-nuclear Factor Kappa B And Inflammatory Factors In Kidney Of High Fat Diet Rats And Intervention Of Liraglutide

Objective:Diabetic nephropathy is a common chronic complication of diabetes mellitus,which is also the important cause of end stage renal disease.Its pathogenesis is complex,in recent years the study found that inflammation plays an important role in DN.TNF-? and IL-6 are the main inflammatory markers,while nuclear transcription factor kappa B is an important mechanism to link obesity and inflammation,which can regulation and transcription the expression of many inflammatory cytokines.DN also has different degrees of glucose and lipid metabolism disorder,and they are often copathogenic,which is also closely related to inflammation.The disorder of lipid metabolism can induce the cell to produce inflammatory reaction,and then cause insulin resistance.Insulin resistance can stimulate the release of variety of inflammatory cytokines,aggravate the glomerular hypertrophy.In addition,hyperinsulinemia by insulin resistsnce brings can also increase glomerular blood flow,glomerular capillary hydrostatic pressure,lead to a high perfusion state and albuminuria.Excessive lipid deposition in the glomerular can lead to glomerular sclerosis.Glucagon like peptide-1 receptor agonist liraglutide is currently new drugs in the treatment of diabetes,studies have found that liraglutide can improve insulin resistance,reduce weight,it also can reduce urinary protein,the glomerular hyperfiltration and renal hypertrophy in diabetic rats then improve renal function,while the mechanism is unclear.In our study,by observing high fat feeding insulin resistance rat renal function,urine protein,blood lipids and the expression of NF kappa B,TNF-? and IL-6 in serum and renal tissue,to explore the expression of kidney inflammatory factors and the correlation with blood lipids in prediabetes,and to explore the effects of liraglutide on renal inflammation process and mechanism after liraglutide intervention,then provide a theoretical basis for its application in diabetic nephropathy and other renal diseases.Method:54 male wistar rats(230-260g)were randomly divided into normal contol group 16 were feed with normal diet and model group 38 were feed with high fat diet.After eight weeks,then 5 rats in each group to calculate glucose infusionrate and evaluate insulin resistance by hyperinsulinemic euglycemic clamp technique.IR rat models were tested successful,then model group was randomly divided into high fat group,liraglutide low dose group(100?g/kg/d)and liraglutide high dose group(200?g/kg/d).11 rats in each group.The first two groups subcutaneous injection with saline);The other two groups subcutaneous injection with liraglutide(100?g/kg/d)and(200?g/kg/d).After 2 weeks of liraglutide intervention,then 5 rats again in each group to calculate GIR by hyperinsulinemic euglycemic clamp technique.The rest of the 6 rats in each group 24 hours urine were collected with metabolic cage then were abdominal aorta bleeding,kidney specimens for transmission electron microscope to observe the renal pathological structure and application of real time PCR technique to determine NF-kappa B,TNF-? and IL-6 m RNA expression in renal tissue.Detection of the serum creatinine and urea nitrogen,total cholesterol,triglyceride,low density lipoprotein cholesterol by tautomatic biochemical analyzer,24 h urine micro albumin was determined by immunoassay.The protein concentration of serum NF-?B,TNF-? and IL-6 was measured by ELISA.Results:1 Comparison of the general situation in four groups of ratsCompared with contol group,the Scr,BUN,MAU,TCH,TG,LDL-C in creased in high fat group(P<0.05);Compared with high fat group,the Scr,BUN,MAU,TG,LDL-C decreased in liraglutide low dose group(P<0.05),the Scr,BUN,MAU,TCH,TG,LDL-C in liraglutide high dose group decreased significantly(P<0.05);Compared with liraglutide low dose group,the Scr,BUN,MAU,TG,LDL-C in liraglutide high dose group decreased further(P<0.05).2 Comparison of GIR in four groups of ratsBefore the drug intervention:Compared with contol group GIR(30.21±2.51mg/kg.min),high fat group GIR(20.41±3.64mg/kg.min)decreased,the difference was statistically significant(P<0.05).After drug intervention for two weeks:Compared with the control group GIR(30.77±2.37mg/kg.min),high fat group GIR(18.83±1.95mg/kg.min)decreased,the difference was statistically significant(P<0.05).After two weeks of liraglutide intervention,compared with the high fat group,the liraglutide low dose group GIR(21.55±2.46mg/kg.min)increased(P<0.05);Compared with the liraglutide low dose group,the liraglutide high dose group GIR(25.80±1.40mg/kg.min)increased further,the difference was statistically significant(P<0.05).3 Comparison of the serum protein concentrations of NF-?B,TNF-? and IL-6 in four groups of ratsCompared with the control group,the serum NF-?B,TNF-? and IL-6 protein levels were increased(P<0.05)in high fat group;Compared with the high fat group,the protein concentrations of the three factors with liraglutide intervention were decreased(P<0.05);and in liraglutide high dose group the protein concentrations of NF-?B,TNF-? and IL-6 decreased further than liraglutide low dose group(P<0.05).4 Comparison of the expression of NF-?B,TNF-? and IL-6 m RNA in kidney of four groups of ratsCompared with the control group,the expression of NF-?B,TNF-? and IL-6 m RNA in kidney in high fat group were increased(P<0.05);Compared with the high fat group,the expression of NF-?B,TNF-? and IL-6 m RNA in kidney were decreased after liraglutide intervention(P<0.05);Compared with liraglutide low dose group,the expression of NF-?B,TNF-? and IL-6 m RNA in kidney decreased further in liraglutide high dose group(P<0.05).5 Comparison of electron microscopy ultrastructure changes of renal tissue in four groups of ratsTransmission electron microscopy showed that(magnified 10000 times): Glomerular structural integrity,Glomerular basement membrane(GBM)clear and podocytic was well distributed,podocytic gap was obvious in control group rats;While in high fat group rats the glomerular podocyte swelling,partial fusion or shedding,and the basement membrane were thickening;Liraglutide intervention can ameliorate the changes above,especially in liraglutide high dose group.6 Correlation analysis resultsThe protein concentration of serum TNF-? and IL-6 was positively correlated with MAU(r=0.802,0.748,P<0.05);The expression of TNF-?m RNA in kidney showed a significant positive correlation with the TCH and TG,(r=0.878,0.817,P<0.05);The expression of IL-6 m RNA in kidney was positively correlated with TCH and TG(r=0.806,0.854,P<0.05);The expression of NF-?B m RNA in kidney was positively correlated with the expression of TNF-? and IL-6 m RNA(r=0.810,0.867,P<0.05).Conclusion:1 High fat diet can cause lipid metabolism disorder and insulin resistance in rats,and can increase the expression of renal NF-kappa B and inflammatory factors TNF-? and IL-6 to activate the inflammatory response,than increase proteinuria,increase kidney damage.2 Liraglutide can dose dependence reduce disorder of lipid metabolism,improve insulin resistance,reduce proteinuria and protect renal function.3 Liraglutide can dose dependence inhibit the expression of renal NF-?B and downregulate the expression of renal inflammatory factor TNF-? and IL-6,improve renal function.