The Effect Of1,25-dihydroxyvitamin D₃on The Expression Of TLR4、 MyD88、NF-κB In Rat With IgA Nephropathy

Objective:Through establish the IgAN rat model, to observe the expression of TLR4、MyD88、NF-κB in rat with IgA Nephropathy and the effect of1,25-dihydro-xyvitamin D3on IgAN rats,and to explore the possible mechanism of1,25-dihydroxyvita-min D3whether they have an protective effect on thedevelopment of IgAN.Method:Select the level of SPF48SD male rats by using bovine serum albumin (BSA)+1ipopolysaeeharide(LPS)+carbon tetrachloride (CCL4), this method which establishan experimental rat model of IgAN(Total9weeks), they have been divided into threegroups randomly:a group of the normal, a group of model and a group of treatment.However,the treatment group was treated with0.05ml peanut oil soluble1,25-dihydroxyvitamin D3administered to0.03ug/kg.d from8week to10weeks,At thesame time point in the model group and the control group were given daily fed withthe same amount of peanut oil.as a result,half of the rats in each group were sacrificedin the13week and the others were killed at the end of the experiment(the17week).,During the experiment, continue observing the following indicators:1.the ratgeneral state.the2.24h urine protein, serum creatinine and other indicators in rats.3.using optical microscopy and immunofluorescence pathology to identificationmodeling success.4.Immunohistochemical detection of renal TLR4,MyD88,NF-κBexpression.5.Semi-quantitative RT-PCR to detect kidney tissue TLR4, MyD88, NF-κB expression.Result:1.the normal group rats were in high spirit by the course of the experiment,reactive activities have no obvious abnormalities,while the model group rats reducedactivities of different levels,apathetic and eating a little,but the treatment group ratssymptoms have eased after treatment.2.the normal group rats did not change significantly at24h urine protein during the experiment, as a matter of fact,the model group and the treatment group havesuccess in modeling,compared with the normal control group the24h urine protein,serum creatinine has been increased,but blood albumin has been decreased,however,the difference was statistically significant (P<0.05).After a treatment of1,25-dihydroxyvitamin D3has declined the average of the24h urine protein, serumcreatinine,only to be found blood albumin increased. Finally the difference wasstatistically significant (P<0.05). Compared with the4weeks of treatment,the8weeks of treatment has changed more evidently. And the difference was statisticallysignificant (P<0.05).3.Pathological optical microscopy shows that the normal control group showedno mesangial cell proliferation,but tubule structure is regular.Therefore there is noinflammatory cell infiltration in renal interstitial.compared with the normal controlgroup,the model group mesangial matrix has increased significantly,mesangial cellproliferation,and there is inflammatory cell infiltration in renal interstitial,immunofluorescence pathology shows that the model group of mesangial area visiblelumpy green fluorescent IgA while the normal control group had no IgA deposition.4.Immunohistochemical results that show that the model group and treatmentgroups kidney tissue TLR4, MyD88, NF-κB expression were higher than the normalcontrol group.because the difference was statistically significant (P<0.05). Comparedwith the model group,the treatment group’s expression of TLR4,MyD88,NF-κB arerelative weak,the difference was statistically significant (P<0.05). Compared with4weeks of treatment,8weeks of treatment’s expression of TLR4, MyD88,NF-κB isweaker,the difference was statistically significant (P<0.05).5.Semi-quantitative RT-PCR results that show after the model of IgAN hassucceed,the expression of TLR4,MyD88,NF-κB were increased significantly,thedifference with the normal control group was statistically significant (P<0.05).Treatment group compared with the model group,the expression of TLR4,MyD88,NF-κ B were decreased,the difference with the model group was statisticallysignificant (P<0.05).Compared with4weeks of treatment,8weeks of treatment’sexpression of TLR4, MyD88,NF-κ B is weaker,the difference was statisticallysignificant (P<0.05). Conclusion:Through increasing the expression of TLR4and MyD88,they could upregulatedto increase inflammatory factors,for example NF-κB，one cause which maybe thedisease progression of IgAN.1,25-dihydroxyvitamin D3decrease the expression ofTLR4, MyD88, downstream effective inflammatory factors,and to reduceproteinuria, serum creatinine lower levels and elevated serum albumin levels,whichwould play a protective role on IgAN.