The Expression Of Apoptosis Associated Proteins In Tonsils Of IgA Nephropathy Patients And Triptolide Intervention Research

Now IgA nephropathy has become the most wide-spread primary glomerulonephritis in the whole world, and it is regarded as the essential reason for the end-stage renal desease(ESRD) as well. The main pathological manifestation is the deposition of IgA in the mesangial area of glomeruli, with or without the deposition of IgM、IgG. It is acknowledged that its pathogenesis has a close relationship with the mucosal immunity, and plenty of research have been done to explore the role that tonsil--the mucosa asscociated lymphoid tissure(MALT) played in the pathogenesis of IgA nephropathy. Bcl-2family proteins are capable to regulate the apoptosis of germinal center of peripheral immune organ through the mitochondrial pathway. The over expression of Bcl-2could even induce the Follicular lymphoma. We speculate the aberrant expression of Bcl-2family proteins might broke the balance between the proliferation and apoptosis in germinal center of tonsil, as well enhance the mucosal immunity. Meanwhile, triptolide might inhibit the mucosal immune response by regulate the expression of Bcl-2family protein in the lymphocytes of tonsils.Objective:Exam the expression of Bcl-2family proteins in the tonsils of patients with IgA nephropathy and regulation of triptolide to the Bcl-2family proteins in the tonsils.Methods:We collect the tonsils from17IgA nephropathy(IgAN) patients and13chronic tonsilitis (CT) patients which were used as control group. Immunohistochemical stain was performed to detect the expression of Bcl-2、 Bcl-xL、Bax in tonsil component; Mononuclear cells were extracted from tonsils of IgA nephropathy group by density-gradient centrifugation, and the expression of Bcl-2、Bcl-xL、Bax in tonsil mononuclear cells were measured by Western-Blot. The separated tonsillar lymphocytes were cultured with0ng/ml、10ng/ml、20ng/ml、30ng/ml of triptolide for72h. The expression of Bcl-2、Bcl-xL、 Bax were examined by Western-Blot.Result:1、The anti-apoptosis proteins(Bcl-2、Bcl-xL) of IgAN group were significantly higher than that of CT group, and they were mainly expressed in the light region of germinal center in tonsil. The pro-apoptosis proteins(Bax) of IgAN group were significantly lower than that of CT group, and they were mainly expressed in the dark region of germinal center in tonsil. The immunohistochemistry result were futher proved by Western-Blot.2、The expression level of Bcl-2of mononuclear significantly negatively correlated with the concentration of triptolide (R=-0.729, P<0.001), and when concentration was20ng/ml or30ng/ml the lower expression level of Bcl-2than the control was statistically significant (P<0.05); The expression level of Bcl-xL of mononuclear significantly negatively correlated with the concentration of triptolide(R=-0.48, P<0.05), and when concentration was20ng/ml or30ng/ml the lower expression level of Bcl-xL than the control was statistically significant (P<0.05); The expression level of Bax of mononuclear significantly positively correlated with the concentration of triptolide (R=0.726, P<0.001), and when concentration was30ng/ml the higher expression level of Bax than the control was statistically significant (P<0.05);Conclusion:Compared to the CT group the balance between the apoptosis and proliferation in the germinal center of tonsils in IgAN group is broken, the apoptosis of lymphocyte is inhibited. This might result in the enhancement of mucosal immunity in tonsil、the more IgAl-produced plasma cells and the deposition of IgA in the mesangial area of glomeruli. Triptolide might influence the apoptosis of tonsil mononuclear cell of IgAN patients through the regulation of Bcl-2family proteins.