电针对佐剂性关节炎大鼠踝关节周围皮肤及皮下组织神经纤维上BSI-B4与IL-1RI荧光双标免疫反应性的影响

目的:研究电针对佐剂性关节炎大鼠踝关节周围皮肤及皮下组织神经纤维上BSI -B4与IL -1RI荧光双标免疫反应性的影响。方法:采用免疫荧光双标技术,观察了佐剂性关节炎大鼠致炎后第3d踝关节周围皮肤及皮下组织神经纤维上BSI- B4与IL 1RI荧光双标免疫反应性,及电针胆经“环跳”穴、“阳陵泉”穴(刺激参数为0 .5～1 .5V ,4～1 6Hz ,30min)后IL 1受体拮抗剂对其是否具有调控作用。结果:①各组大鼠炎症侧外踝关节周围皮肤及皮下组织均可见BSI B4 /IL -1RI阳性双标纤维。②致炎后第3d ,炎症组大鼠外踝关节周围皮肤及皮下组织BSI- B4阳性纤维、IL -1RI阳性纤维、IL- 1RI/BSI B4双标纤维数及双标纤维占各单标纤维的百分比较生理盐水组显著升高(P <0 .0 1 ,P <0 .0 5 )。③IL -1受体拮抗剂组大鼠BSI B4阳性纤维、IL -1RI阳性纤维、IL -1RI/BSI B4双标纤维数及双标纤维占IL- 1RI单标纤维的百分比较炎症组显著降低(P <0 .0 1 ,P <0 .0 5 )。④电针组大鼠BSI B4阳性纤维及IL- 1RI/BSI B4双标纤维数、双标纤维占IL -1RI单标纤维的百分比显著低于炎症组(P <0 .0 5 ) ,并高于IL -1受体拮抗剂组(P <0 .0 1 )。结论:正常大鼠踝关节周围皮肤及皮下组织C纤维上有IL- 1RI表达,完全弗氏佐剂导致的踝关节周围局部炎症组织中该表?

Effect of Electroacupuncture on the Number of BSI-B4/IL-1R Double Labeled Nerve Fibers in the Dermal and Subcutaneous Tissues around the Ankle Joint in Adjuvant Arthritis Rats

Objective:To observe the effect of electroacupuncture (EA) on the number of Bandeiraea Simplicifolia I (BSI)-B4 labeled and IL-1 receptor I immunoreaction (IR) positive nerve fibers of the dermal and subcutaneous tissues around the ankle joint in adjuvant monoarthritis rats. Methods: 28 male SD rats (180～200 g) were randomized into control group, model group, IL-1 receptor antagonist (IL-1-RA) group and EA group, with 7 cases being in each group. Adjuvant monoarthritis model was induced by injection of 50 μL complete Freund's adjuvant (CFA) into the left hind ankle joint cavity. EA (4～16 Hz, 0.5～1.5 V and intermittent waves) was applied to “Huantiao”(GB 30) and “Yanglinquan”(GB 34) for 30 min following injection of CFA and on the 3rd day. In IL-1-RA group, on the 1st and 3rd day after injection of CFA, IL-1RA (1 μg/mL, 50 μL) was injected into the subcutaneous tissues around the left hind ankle joint. On the 3rd day, the rats were anesthetized with 20% urethane (1 g/kg) and transcardiac perfusion was performed by using 4% paraformaldehyde phosphate buffer (pH 7.4). The skin and subcutaneous tissues around the focus region were taken to be cut into sections (10 μm) which were then treated with BSI-B4 staining (for displaying C-fibers) and IL-1RI immuno-fluorescent labeling techniques. The number of BSI-B4/IL-1RI double labeled IR-positive nerve fibers (30 sections/rat,38.44 mm2/section) was calculated and analyzed by using fluorescent microscope and image analysis system. Results: 1) The double labeled (BSI-B4/IL-1RI/IR positive) fibers mainly distributed in the dermal and subcutaneous tissues around the ankle joint. 2) Compared with control group, BSI-B4 labeled fibers and IL-1RI/IR-positive fibers, and the double labeled fibers and the percentage of double labeled fibers/ BSI-B4 or IL-1RI single-labeled fibers in model group, IL-1-RA and EA groups were significantly higher(P<0.05～0.01); 3)In comparison with model group, the number of BSI-B4 labeled fibers, IL-1RI IR-positive fibers, double stained fibers and the percentage of double-stained fibers/IL-1RI single-stained fibers of IL-1-RA group was markedly lowered(P<0.05～0.01; 4)Compared with IL-1-RA group, the number of BSI-B4 labeled fibers, double labeled fibers and the percentage of double-labeled fibers / IL-1RI IR single-stained fibers of EA group was markedly lowered(P<0.05), and the percentage of double labeled fibers/BSI-B4 labeled fibers was significantly higher(P<0.05). Conclusion: 1) There is IL-1RI expression in the C-fibers of the dermal and subcutaneous tissues around the ankle joint in normal rats, which is upregulated in the inflammatory tissue induced by CFA. The analgesic effect of EA is probably related to its suppressive effect on C-fibers' proliferation and IL-1β(inflammation factor) induced activation of IL-1RI and its promotion effect on the synthesis of IL-1RA (the endogenous anti-inflammatory factor) in the inflammation focus.