风寒湿外邪对痹证(佐剂性关节炎)发生发展的影响

Effect of Exogenous Evils of Wind-cold-damp on the Occurrence and Development of Bi Syndrome(Adjuvant-induced Arthritis)

目的探讨风寒湿外邪对痹证[类风湿关节炎]发生发展的影响。方法 (1)雄性SD系大鼠20只,采用分层随机法分为单纯完全弗氏佐剂(CFA)组(10只)及风寒湿14天+CFA组(10只)。单纯CFA组大鼠正常饲养14天,风寒湿+CFA组大鼠风寒湿刺激14天,尾根部皮下注射0.1mL含100μg结核分支杆菌(Mtb)的CFA诱导佐剂性关节炎(AIA)观察并记录大鼠发病数,计算发病率。(2)雄性SD系大鼠30只,分层随机分为单纯CFA组(10只)、风寒湿7天+CFA组(10只)及风寒湿14天+CFA组(10只),3组大鼠分别尾根部皮下注射0.1mL含200μg Mtb的CFA诱导AIA。观察各组大鼠发病时间及关节炎指数。(3)雄性SD系大鼠30只,分层随机分为正常组(10只)、单纯CFA组(10只)及风寒湿14天+CFA组(10只),观察并记录各组大鼠精神状态、舌质、大便情况,测量体质量、饮水量和光辐射热刺激撤足时间。(4)雄性SD系大鼠20只,分层随机分为正常组(10只)及风寒湿14天组(10只),检测外周血CD4+T细胞亚群。结果 CFA(含100μg Mtb)注射后,单纯CFA组大鼠关节炎发病率为50%,风寒湿刺激14天+CFA组大鼠达到100%(P=0.039)。风寒湿刺激14天+CFA组在CFA(含200μg Mtb)注射后出现关节炎时间为(8.3±1.2)天,比单纯CFA组大鼠[(13.2±1.2)天]显著提前(P<0.01);与单纯CFA组比较,风寒湿14天+CFA组大鼠体毛欠光泽、明显畏寒、蜷卧懒动、大便稀溏,舌质偏淡、舌苔腻。风寒湿14天+CFA组大鼠关节炎初发阶段体重显著下降(P<0.05,P<0.01),后足肿胀程度显著增加(P<0.05,P<0.01),饮水量显著减少(P<0.05),热刺激撤足时间显著增加(P<0.01)。风寒湿14天组大鼠外周血中CD4+T细胞亚群数量较正常组明显升高(P<0.05)。结论风寒湿外邪影响AIA病证的发生,在疾病初发阶段中医证候表现为风湿寒痹证,其形成机制可能与CD4+T细胞亚群异常活化有关。

Objective To observe the effects of exogenous evils of wind-cold-damp on the oc- currence and development of Bi syndrome （rheumatoid arthritis, RA）. Methods （1）Twenty male SD rats were randomly divided into Group A [complete Freund＇s adjuvant （CFA） alone] and Group B （14 days wind-cold-damp stimulation ＋CFA）, 10 in each group. Rats in Group A were routinely fed for 14 days.Rats in Group B received wind-cold-damp stimulation for 14 days, 0.1 mL CFA comtaining 100 μg Myco- bactrium tuberculosis （Mtb） was subcutaneously injected to rats in the 2 groups from tail root to induce adjurant-induced arthritis （AIA）. The morbidity was daily observed and recorded, and the incidence was calculated. （2）A total of 30 male SD rats were randomly stratified into Group C （CFA alone）, Group D （7 days wind-cold-damp stimulation ＋ CFA）, Group E （14 days wind-cold-damp stimulation）, 10 in each group. 0.1 mL CFA containing 200 μg Mtb was subcutaneously injected to rats in the 3 groups from tail root to induce AIA. The onset time and arthritis index （AI） were observed.（3） A total of 30 male SD rats were randomly stratified into the normal group （F）, Group G （CFA alone）, Group H （14 days wind-cold- damp stimulation ＋CFA）, 10 in each group. The psychosis, tongue proper, defecation were observed and recorded in each group. Body mass index （BMi）, water consumption, and thermal hyperalgesia in hind paws of rats were measured. （4） Twenty male SD rats were randomly stratified into the normal group （I） and Group J （14 days wind-cold-damp stimulation）, 10 in each group. Peripheral CD4 ＋ T cell subsets were detected by flow cytometry. Results （1） After injection of CFA （containing 200 μg Mtb）, the incidence of AIA was 50% in Group A and 100% in Group B （P =0.039）. （2） Arthritis occurred in Group E at day （8.3 ±1.2） day after injection of CFA （containing 200 μg Mtb）, earlier than that of Group C [ （13.2 ±1.2） days, P 〈0.011. （3） Compared with AIA rats normally fed, rats in Group H had no luster hair but chilliness. They were lazy to move. They had loose stool, pale tongue, and greasy fur. Body weight significantly decreased at the initial stage of arthritis in Group H （P 〈0.05 ,P 〈0.01 ）. The swelling degree of hind toes significantly increased （P 〈0.05 ,P 〈0.01 ）, water consumption was significantly re- duced （P 〈0.05）, thermal stimulation pain threshold significantly increased （P 〈0.01 ）. （4） The number of CD4＋ T cells subsets in the peripheral blood of rats in Group J was obviously more than that of Group I （P 〈0.05）. Conclusions Exogenous evils of wind-cold-damp affect the occurrence of AIA, which might manifest as wind-cold-damp Bi syndrome at the initial stage. Its pathogenesis might be associated with abnormal activation of CD4 ＋ T cell subsets.