Study On The Regulation Of PPAR-γ/NF-κB Pathway In The Treatment Of IgA Nephropathy By Replenishing Qi, Invigorating Spleen And Removing Dampness

ObjectiveObjective To observe the effects of replenishing qi,invigorating spleen and removing dampness and peroxisome proliferator activated receptor-γ(PPAR-γ)agonists on 24-hour urinary protein,renal function and renal pathological changes in IgA nephropathy rats,and to compare the expression of key proteins of nuclear transcription factor-κB(NF-κB)signal pathway in renal tissue,so as to preliminarily explore the effect of replenishing qi,invigorating spleen and removing dampness on IgA nephropathy.It provides a new idea for the clinical treatment of IgA nephropathy.MethodsThirty two SD rats,6 weeks old,weighing about 180 g,were used to randomly divid into control group,IgA nephropathy model group,Yi-Shen-Hua-Shi granule group and rosiglitazone group.The animal model was induced by the combination of lipopolysaccharide,bovine serum albumin and carbon tetrachloride.In the modeling period(the 1-8 weeks of the experiment):control group: from the beginning of the experiment,600 mg/kg distilled water was given by gavage every two days until the end of the eighth week.The rats were injected subcutaneously with 0.4 ml normal saline once a week until the end of the 8th week.At the 6th and 8th week of the experiment,each rat was injected with 0.2ml normal saline via tail vein once a week.The other three groups: from the beginning of the experiment,600 mg/kg bovine serum albumin was given by gavage every two days.0.1ml carbon tetrachloride and 0.3ml castor oil were injected subcutaneously once a week until the end of the 8th week.At the 6th and 8th week of the experiment,each rat was injected with 0.05 mg lipopolysaccharide via tail vein once a week.In the treatment period(9-12 weeks): control group and model group: at the beginning of the9 th week,2ml distilled water was given by gavage daily until the end of the 12 th week.YiShen-Hua-Shi granule group: at the beginning of the 9th week,2.7g/kg Yi-Shen-Hua-Shi granule was given by gavage daily until the end of the 12 th week.Rosiglitazone group: 0.72mg/kg rosiglitazone was given daily from the 9th week until the end of the 12 th week.The general state,24-hour urine protein,IgA deposition in kidney and the protein expression of NF-κB P65,P NF-κB P65 and PPAR-γ were observed.Results1.The mental state,glossiness,activity,diet and stool of the blank control group were all well performed.The rats in model group showed mental poor,decreased food,no activity,decreased glossiness and yellow color,and the weight of the rats in the model group was lower than that in the blank group.When carbon tetrachloride was injected subcutaneously,the subcutaneous sclerotia appeared in the model group,and the hair removal was serious.And after the injection of lipopolysaccharide,there will be more serious mental loss,loss of appetite,and reduced activity.The mental state,activity and diet of rats in Yi-Shen-Hua-Shi granule group and rosiglitazone group were improved after receiving drug treatment.However,after treatment,the rats in the traditional Chinese medicine group had loose stools.The appetite of rosiglitazone group was stronger.2.The results of 24-hour urinary protein showed that the 24-hour urinary protein of YiShen-Hua-Shi granule group and rosiglitazone group were lower than that of model group after treatment.Compared with the model group,the 24-hour urinary protein in Yi-Shen-Hua-Shi granule group was significantly different(P = 0,P < 0.01).These results suggest that replenishing qi,invigorating spleen and removing dampness can reduce 24-hour urinary protein in IgA nephropathy rats.3.The results of renal function showed that after treatment,the urea nitrogen and serum creatinine of Yi-Shen-Hua-Shi granule group and rosiglitazone group were lower than those of model group,and the renal function of Yi-Shen-Hua-Shi granule group was significantly different from that of model group,which suggested that replenishing qi,invigorating spleen and removing dampness could reduce the renal function of IgA nephropathy rats.4.The results of IgA immunofluorescence staining showed that the scores of IgA immunofluorescence staining in each group(except the blank control group)were mostly between+～++.The Yi-Shen-Hua-Shi granule group and rosiglitazone group could significantly alleviate the IgA deposition in the glomerular mesangial area of the model group.HE staining and Masson staining results showed that the renal tissue of the model group showed different degrees of pathological changes.After treatment,compared with the model group,Yi-ShenHua-Shi granule group and rosiglitazone group can significantly reduce the pathological changes of IgA nephropathy rats,such as improving the proliferation of glomerular mesangial cells and the increase of matrix.5.According to Western blot analysis,the protein expression of NF-κB P65 and pNF-κB P65 in Yishenhuashi granules group and rosiglitazone group were significantly decreased compared with that in model group,while the expression of PPAR-γ in Yi-Shen-Hua-Shi granule group and rosiglitazone group had no statistical significance(P>0.05).These results suggest that the method of replenishing qi,invigorating spleen and removing dampness may be used to treat IgA nephropathy by regulating PPAR-γ/NF-κB pathway.Conclusion1.The method of Replenishing qi,invigorating spleen and removing dampness can effectively reduce the 24-hour urinary protein level of rats with IgA nephropathy,improve the renal function of rats with IgA nephropathy,improve the renal pathological status of rats with IgA nephropathy,and delay the progression of IgA nephropathy.2.The method of Replenishing qi,invigorating spleen and removing dampness can inhibit the activation of renal inflammatory pathway in rats with IgA nephropathy,and reduce the expression of P65 and p-p65.The method of invigorating qi,invigorating spleen and dispiriting dampness may be used to treat IgA nephropathy by regulating PPAR-γ/NF-κB pathway.