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目的 探讨雷公藤内酯醇 (TP)体内外对致敏大鼠淋巴细胞凋亡的影响。方法 采用卵蛋白 (OVA)致敏并反复刺激建立过敏性气道炎症模型 ,2 4只SD大鼠随机分为正常组、阳性对照组和TP处理组 ,每组 8只。用TUNEL原位末端标记法 ,DNA电泳及电镜等方法 ,观察体内外TP对致敏大鼠淋巴细胞凋亡的影响及机制。结果 致敏大鼠BALF中嗜酸性粒细胞 (Eos)、淋巴细胞均较正常组明显增高 (P <0 .0 5 )。体内应用TP可减少致敏大鼠BALF中Eos、淋巴细胞数目 ,同时可增加其BALF中淋巴细胞凋亡百分率。体外实验显示 ,不同剂量TP呈剂量依赖性 (10 7～ 10 5 g ml)的促进OVA抗原刺激的脾淋巴细胞凋亡 ,该效应随作用时间延长而增强 ,并促进Fas FasL表达 ;DNA电泳呈现梯形带 ,凋亡细胞呈典型凋亡改变 ;10 7g mlTP可明显促进 10 6 mol L地塞米松 (DXM)的促凋亡作用。结论 TP可明显促进致敏大鼠肺及脾淋巴细胞凋亡 ,可能是其抗炎机制之一 ,并可能是通过Fas FasL途径发挥作用 ;同时TP可明显增加DXM的促淋巴细胞凋亡作用。为阐明TP对抗哮喘气道炎症的作用机制和探讨激素依赖性哮喘治疗的新途径提供了有意义的实验资料。
Objective To investigate the effects of triptolide (TP) on lymphocyte apoptosis in sensitized rats in vitro and in vivo. Methods Allergic airway inflammation model was induced by ovalbumin (OVA) and repeatedly stimulated. 24 SD rats were randomly divided into normal group, positive control group and TP treatment group, with 8 rats in each group. TUNEL in situ terminal labeling method, DNA electrophoresis and electron microscopy and other methods to observe in vitro and in vivo effects of TP on apoptosis of lymphocytes in sensitized rats and its mechanism. Results The eosinophils (Eos) and lymphocytes in the BALF of sensitized rats were significantly higher than those in the normal group (P <0.05). The in vivo application of TP can reduce the number of Eos and lymphocytes in the BALF of sensitized rats and increase the percentage of lymphocyte apoptosis in BALF. In vitro experiments showed that TP dose-dependently (10 7 ~ 10 5 g ml) promoted the apoptosis of splenic lymphocytes stimulated by OVA antigen, which increased with the prolongation of time and promoted the expression of Fas FasL. DNA electrophoresis showed Apoptotic cells showed typical apoptotic changes in trapezoidal bands. 10 7 g mlTP could obviously promote the apoptosis of dexamethasone (DXM). Conclusion TP can significantly promote the apoptosis of lung and spleen lymphocytes in sensitized rats, which may be one of its anti-inflammatory mechanisms and may play a role through the Fas FasL pathway. At the same time, TP can obviously increase the lymphocyte apoptosis induced by DXM. In order to elucidate the mechanism of action of TP on asthma airway inflammation and to explore new ways of hormone-dependent asthma treatment, we provided meaningful experimental data.