雷公藤内酯醇对大鼠胶原性关节炎和免疫功能的影响

Effect of triptolide on collagen-induced arthritis and immune function in rats

目的研究雷公藤内酯醇(TP)对胶原性关节炎(CIA)的治疗作用。方法制备鸡Ⅱ型胶原诱导的CIA大鼠模型,im给予地塞米松2.0 mg·kg-1或TP 0.1,0.2和0.4 mg·kg-1,每3天1次,连续35 d,正常对照和模型对照组im给予等体积生理盐水。实验过程中观察CIA大鼠的一般情况、足爪肿胀度和关节炎指数的变化。给药结束后第2天,3%巴比妥钠麻醉处死大鼠,取足爪组织制备组织切片观察病理变化;无菌取脾制备脾细胞悬液,用MTT法检测脾T细胞和B细胞增殖反应;制备10%脾组织匀浆,用ELISA法检测脾组织白介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)和IL-6;心脏取血,制备血清用ELISA法测定血清中抗CⅡ抗体水平。结果与正常对照组比较,模型对照组大鼠关节红肿,表面皮肤发亮充血,关节炎指数上升,脾T细胞和B细胞增殖反应、血清中抗CⅡ抗体水平和脾组织中IL-1β,TNF-α和IL-6水平明显升高(P<0.01),大鼠足爪皮下组织细胞排列紊乱且有大量炎性细胞浸润及血管增生。im给予TP 0.2和0.4 mg·kg-1可改善CIA大鼠一般状况,明显抑制CIA大鼠足爪肿胀(P<0.01);降低关节炎指数,在第22,26,30和35天由模型对照组的7.3±0.7,6.3±0.8,4.7±0.7和4.3±0.2分别降低到5.6±0.6,4.9±0.7,3.9±0.5,2.8±0.7和5.1±0.8,4.1±0.4,3.3±0.6,2.7±0.6;抑制脾T和B细胞增殖反应;血清中抗CⅡ抗体水平显著降低,A490 nm由模型对照组的0.765±0.018降低到0.583±0.024和0.575±0.034(P<0.01);脾组织中IL-1β由(159±23)ng·L-1降低到90±15和(79±15)ng·L-1(P<0.01),TNF-α由(125±23)ng·L-1降低到94±16和(83±17)ng·L-1(P<0.01),IL-6由(250±16)ng·L-1降低到193±18和(180±18)ng·L-1(P<0.01);能明显改善大鼠足爪组织病理改变,炎性细胞浸润及血管明显减轻(P<0.05)。TP 0.1 mg·kg-1对上述指标无明显影响。结论 TP对Ⅱ型胶原诱导的CIA具有一定的治疗作用。

OBJECTIVE To investigate the effect of triptolide （TP） on collagen-induced arthritis （CIA）. METHODS Chicken type Ⅱcollagen （ C Ⅱ） was used to induce the CIA model. Dexamethasone 2.0 mg·kg^-1 or TP 0.1, 0.2 and 0.4 mg·kg^-1 was im given to rats, once for 3 d, until 35 d. The change of the rats, the paw swelling degree and the articular index （AI） were observed. On the 3 5th day, the rats were sacrificed and the histopathological changes in paw tissues were observed by HE staining. The proliferation of spleen T and B lymphocytes was detected by MTT assay. The level of interleukin-1 β （ IL-1 β, TNF-et and IL-6 in spleen tissue homogenate and anti- C Ⅱ IgG antibody in serum was examined by ELISA. RESULTS Compared with normal control group, the joint of rats in model group showed swelling, the surface of skin shined and congested, and AI rose. The proliferation of spleen T and B lymphocytes, the level of anti-C Ⅱ IgG antibody in serum and the content of IL-113, TNF-tx and IL-6 in the spleen all significantly increased （P 〈 O. O1 ）. The subcutaneous tissue cells of the rat and blood vessels proliferated. TP O. 2 and O. 4 paw became deranged, inflammatory cells infiltrated rag-kg-1 was able to improve the general situation of CIA rats, significantly inhibit the paw swelling degree and reduce AI. AI on the 22nd, 26th, 30th and 35th day was reduced from 7.3 ±0.7, 6.3 ±0.8, 4.7 ±0.7 and 4.3 ±0.2 in model group to 5.6 ±0.6, 4.9 ±0.7, 3.9 ±0.5 and 2.8 ±0.7 for TP 0.2 mg·kg^-1, and 5.1 ±0.8, 4.1 ±0.4, 3.3 ±0.6 and 2.7 ±0.6 for TP 0.4 mg·kg^-1 （ p 〈 0.01 ）. The proliferation of spleen T and B lymphocytes was also suppressed. In TP 0.2 and 0.4 mg·kg^-1 groups, the level of anti-C Ⅱ IgG antibody in serum （A490 nm） was reduced from 0. 765 ±0. 018 in model group to 0. 583 ±0. 024 and 0. 575 ±0. 034 （P 〈 0.01 ）, IL-1β content in spleen tissue was reduced from （159 ±23 ）ng· L^-1 to 90 ±15 and （79 ±15）ng＇L^-1 TNF-α from （125 ±23）ng＇L-l to 94 ±16 and （83 ±17）ng·L^-1, and IL-6 from （250±16）ng· L^-1 to 193 ＋18 and（ 180 ±18）ng· L^-1 （P〈0.01）. TP0.2 and 0.4 mg·kg^-1 could reduce the inflammatory cell infiltration and angiogenesis of rat paws. TP 0.1 mg· kg^-1 had no significant effect on the above indicators. CONCLUSION TP can improve the immune function of CIA model rats. TP has therapeutic effect on CIA.