大剂量免疫球蛋白对免疫性周围神经病治疗机理的实验研究

Experimental study on the therapeutic mechanism of high dose intravenous immunoglobulin in treatment of immune-mediated peripheral neuropathy

目的 探讨静脉注射免疫球蛋白 (IVIg)对空肠弯曲菌脂多糖 (CJLPS)诱导的免疫性周围神经病的治疗机理。方法 ( 1)在CJLPS免疫大鼠后的不同时间 ,按 40 0mg/ (kg·d)注射IVIg。第3 5天取坐骨神经进行病理学检查 ,ELISA方法检测血清中抗CJLPS抗体滴度 ,免疫组化及原位杂交技术检测神经上特异性IgG及TNF αmRNA表达 ;( 2 )在CJLPS刺激下体外培养外周血单核细胞(PBMC) ,并分别在培养液中加入 1、2 5、5、10mg/mlIVIg。将上清分别注入同种大鼠的神经外膜下 ,7d后进行病理学检查 ;采用原位杂交技术检测培养PBMC的TNF αmRNA表达。结果 ( 1)IVIg干预下 ,经CJLPS免疫大鼠神经原纤维病变率仅为 1 0 % ,未干预者达 15 0 % ,差异有显著性 (P <0 0 1)。未干预组血清中抗CJLPS特异性IgG滴度比干预组高 9倍。IVIg干预组神经上无明显特异性IgG及TNF αmRNA表达 ,而未用IVIg者表达强阳性 ;( 2 )CJLPS免疫的大鼠PBMC的TNF αmRNA ,IVIg干预组 ( 1 0 % )比IVIg未干预免疫组 ( 9 5% )减少 ;( 3 )CJLPS刺激下健康鼠PBMC体外培养 ,5mg/ml和 10mg/ml大剂量IVIg干预组的TNF αmRNA阳性表达率 ( 3 %和 2 % ) ,较 2 5mg/ml和 1mg/ml小剂量组阳性率 ( 13 %和 11 5% )为低 (P <0 0 1)。其上清液神经外膜下注射后 ,大剂量干?

Objective To explore the therapeutic basis of high dose intravenous immunoglobulin (IVIg) in treatment of peripheral neuropathy induced by Campylobacter jejuni lipopolysaccharide (CJ LPS). Method (1) IVIg (400 mg/kg·d) was given to the rats on the different days respectively during the immunization with CJ LPS. Histological study of sciatic nerve was performed on the 35 th day after immunization. The titer of anti CJ LPS antibody in sera of immunized rats was measured by ELISA; IgG deposition was detected by immunohistochemistry and expression of TNF α mRNA in the pathological nerves by in situ hybridization histochemistry. (2) When PBMCs were stimulated by CJ LPS in vitro, IVIg was added into culture medium at the doses of 1, 2.5, 5, and 10 mg/ml, respectively. Pathological examination of sciatic nerve was performed on the 7th day after perineural injection of the supernatants. Expression of TNF α mRNA in PBMCs stimulated by CJ LPS in medium was detected by in situ hybridization histochemistry after adding IVIg. Results (1) The rate of abnormal fibers appearance in IVIg group (1.0%) was much lower than that of the control group (15.0%) after immunization with CJ LPS, P <0.01. The titer of antibody in control group was 9 times higher than that of IVIg group. There was no expression of immunoglobulin and TNF αmRNA in peripheral nerves in IVIg group, but high expression was found in control group in which no IVIg was injected. (2) The expression rates of TNF αmRNA on the PBMCs in IVIg group (1.0%) was much lower than that of control group (9.5%). (3) When the PBMCs of normal rats were stimulated by CJ LPS, the expression rates of TNF αmRNA in PBMCs of 5 mg/ml IVIg group (3.0%) or 10 mg/ml IVIg group (2.0%) were much lower than that of 1 mg/ml IVIg group (15.0%) or 2.5 mg/ml IVIg group (11.5%), P <0.01. The rate of abnormal fibers appearance in 5 mg/ml IVIg group (9.8%) or 10 mg/ml IVIg group (8.5%) was much lower than that of 1 mg/ml IVIg group (50.0%),2.5 mg/ml IVIg group (41.0%) or control group (50.8%) after the perineural injection with the supernatants, respectively, P <0.01. Conclusion The therapeutic effect of high dose IVIg might be associated with inhibition of the humoral and cellular immunity simutaneously in peripheral neuropathy induced by CJ LPS.