The Research On Expression Level Of Wnt3a/β-catenin Signaling Pathway In Kidney Tissues Of Lupus Nephritis Mice

Objective The expression level in Wnt3a/β-catenin signaling pathway of Wnt3 a and β-catenin in the kidney tissues of the mouse model(MRL/lpr mice)was conducted to determine whether the abnormal expression of Wnt3a/β-catenin signaling pathway has any effect on the occurrence and development of lupus nephritis,which was supposed to add more theoretical evidence for research of molecular biological mechanisms in lupus nephritis.Methods 1.Raising and grouping of the mouse modelThe 15 MRL/lpr female mice purchased 6-week old are grouped as mouse model,and C57BL/6 female mice the same old are grouped as normal model.After 10 weeks,keep these two group of mice in the metabolic cages,and collect their urine as samples.Then under anaesthesia,take out the mice’s eyes by blood draining,sampling their blood,and take out their kidney tissues.2.General biochemical tests on lupus nephritisThe application of enzyme-linked immunosorbent assay(ELISA)was used for examining the number of antinuclear antibody and Anti-double stranded DNA antibody(anti-ds DNA)in the blood of the mice grouped,respectively,and a 24-hour urine protein test was done on the samples analyzed by an automatic biochemical analyzer.3.Pathology tests on the targeted miceHematoxylin-eosin(HE),periodic acid-silver methenamine(PASM),and Masson staining were performed on kideney tissues taken from the experimented mice to observe changes between two tissues from mice of the two groups,respectively.Then immunofluorescence technique was adopted to detect the deposition of immune complexes of Ig G and complement component 3(C3)in the tissues of the mice.4.Tests on Wnt3 a and β-catenin genes of taken kidney tissues by Quantitative real-time PCRQuantitative real-time PCR(qPCR)was applied to examine the expression level of Wnt3 a and β-catenin genes under the comparison between the model mice and normal mice.5.Tests on Wnt3 a and β-catenin of taken kidney tissues by Western blotWestern blot(WB)is adopted to determine the expression of Wnt3 a and β-catenin under the comparison between the mouse model and normal mice.Result 1.General status of the mouse model affected with lupus nephritisIn the mouse model,red rash,hair loss and crusting in the neck appeared when they were 16 weeks old.The splenomegaly was found in the model group compared with the normal control group.2.General biochemical tests on lupus nephritis miceCompared with the normal control group,the(24-hour)urinary protein quantity of the model group was surprisingly higher than that of the control group,and the difference was statistically significant(P<0.05).The serum levels of ANA and ds DNA antibodies in the model group were also higher than those in the normal control group(P<0.05),which was statistically significant.3.Pathological changes in lupus nephritisPathology and immunofluorescence tests both presented that Mesangial cell proliferation,inflammatory cell infiltration,and immune complexes Ig G,C3 deposition were found in the glomeruli in the mouse model.4.q PCR for detection of the expression of Wnt3 a and β-catenin genes in kidney tissues of lupus nephritis miceThe expression level of Wnt3a(2.49 ± 0.939)and β-catenin(1.53 ± 0.110)genes in the kidney tissues of the mouse model were obviously higher than the Wnt3a(0.90±0.156)and β-catenin(0.82 ±0.109)of those in the normal control group,and the difference bears statistical significance(P <0.05).5.WB for detection of the expression of Wnt3 a and β-catenin in kidney tissues of lupus nephritis miceThe expression level of Wnt3 a protein(0.46 ±0.193)in the kidney of the mouse group was particularly higher than that of the normal control group(0.16±0.057),and the difference was statistically significant(P<0.05).The expression level of β-catenin protein(1.59±0.471)in the kidney of the mouse group was particularly higher than that of the normal control group (0.55±0.209),and the difference was statistically significant(P<0.05).Conclusion The signaling pathway of Wnt3a/β-catenin should be in an abnormal activation state,and that the signaling pathway were involved in the development of lupus nephritis,overexpression of which may be one of the molecular biological mechanisms of lupus nephritis.