An Experimental Study On The Expression And Correlativity Of HSP90, Autoantibodies To HSP90 And IL-6, IL-10 In Patients With Systemic Lupus Erythematosus

【Objective】To study the expression and correlativity of HSP90 gene and IL-6 in patients with systemic lupus erythematosus (SLE) through detecting the expression of HSP90 gene in the peripheral blood mononuclearcells (PBMC) and IL-6 in the serum from SLE patients.【Methods】The methods of immunohistochemistry and in situ hybridization technique were used to detect the expression of HSP90 protein and HSP90 mRNA respectively in the PBMC from 45 SLE patients and 15 healthy controls. Semi-quantitative measurement of positive expression in the blood specimen was done with High Multiply Image Analytical System(HMIAS). A solid-phase sandwich ELISA and Western blotting were used to detect the expression of IL-6, IL-10 and antibodies to heat shock protein 90 (HSP90)in the serum in the same groups. Applying linear association test to analyze correlated data.【Results】⑴. In serum of 45 SLE patients, the overexpression of IL-6 level was dected in 31 cases(68.89%). The levels of IL-6 in sera of active SLE patients, inactive patients and healthy people was 16.82±1.79pg/ml, 12.58±1.12pg/ml and 10.13±2.95pg/ml respectively. The overexpression of IL-10 was dected in 29 cases(64.44%). The levels of IL-10 in sera of active SLE patients, inactive patients and healthy people was 47.31±16.20pg/ml, 42.78±11.21pg/ml, 22.05±8.82pg/ml respectively. Active SLE patients higher levels of IL-6 and IL-10 in sera compared with healthy controls and inactive SLE patient(sP<0.01), and inactive SLE patients also had higher levels of IL-6 and IL-10 compared with healthy controls(P<0.05).⑵. Out of 45 SLE patients, the expression of HSP90 protein in PBMC was detected in 17 cases(37.78%). No expression of HSP90 protein was observed in healthy group. Semi-quantitative measurement of HSP90 protein expression in the blood specimen showed that the mean luminosity in active SLE patients, inactive patients and healthy people was 0.412±0.230, 0.187±0.019 and 0.117±0.033 respectively. Active SLE patients had higher levels of HSP90 protein expression in PBMC compared with healthy controls and inactive SLE patients (P<0.01), and inactive SLE patients also had higher levels of HSP90 protein expression compared with healthy control(sP<0.05). The expression of HSP90 mRNA in PBMC was detected in 20 cases(44.44%)in the same group. No expression of HSP90 mRNA was observed in healthy group. Semi-quantitative measurement of HSP90 mRNA expression in the blood specimen showed that the mean luminosity in active SLE patients, inactive patients and healthy people was 0.350±0.225, 0.221±0.040 and 0.114±0.009 respectively. Active SLE patients had higher levels of HSP90 mRNA expression in PBMC compared with healthy controls(P<0.01)and inactive SLE patients (P<0.05), and inactive SLE patients also had higher levels of HSP90 mRNA expression compared with healthy controls(P<0.05). The HSP90 protein level in patients with positive anticardiolipin antibodies was higher compared with that in negative patients.⑶. A linear positive relationship was found between expression of IL-6 and levels of HSP90 mRNA (n=45, r=0.413, t=2.971, P<0.01) and between expression of IL-10 and levels of HSP90 mRNA (n=45, r=0.334, t=2.319, P<0.05). A linear positive relationship was found between expression of IL-6 and levels of HSP90 protein (n=45, r=0.63, t=5.339, P<0.001). There was no relationship was found between expression of IL-10 and levels of HSP90 protein.⑷. Out of 45 SLE patients, the expression of IgG antibodies to HSP90 in serum was detected in 23 cases (51.11%), the expression of IgM antibodies to HSP90 in serum was detected in 11 cases (24.44%). No expression of IgG/IgM antibodies to HSP90 was observed in healthy group.⑸. In SLE patients, elevated levels of HSP90 were significantly correlated with the presence of elevated levels of IgG antibodies to HSP90(X2=4.15,P<0.05) but not with elevated levels of IgM antibodies to HSP90.【Conclusions】The cytokine IL-6 expression in SLE patients is up regulated, which can induce enhanced transcription of the HSP90 gene and in turn results in alevated levels of HSP90 protein. The mass collecting of HSP90 protein can be identified by antigen specific lymphocytic cell and results in the production of antoantibodies to HSP90, Which can result in immunological function disturbance in SLE patients. Which might play a certain role in the pathogenesis of SLE.