目的 初步探讨雷公藤多苷片免疫抑制作用及其HPLC指纹图谱间的相关性。方法 采用HPLC建立7个厂家生产的雷公藤多苷片醋酸乙酯提取物的指纹图谱,选择刀豆蛋白A(Con A)刺激的小鼠脾淋巴细胞模型进行免疫抑制药效学研究,分别考察雷公藤多苷片对小鼠脾淋巴细胞增殖以及细胞因子γ干扰素(IFN-γ)分泌的影响;采用偏最小二乘回归分析法分析谱效关系。结果 建立了7个厂家生产的雷公藤多苷片的HPLC指纹图谱,从HPLC指纹图谱中提取出18个能够表征差异的特征峰。药效研究结果显示不同厂家生产的雷公藤多苷片均可显著抑制Con A诱导的小鼠脾淋巴细胞增殖和IFN-γ的分泌(P<0.05)。偏最小二乘回归法分析结果显示,以抑制小鼠脾淋巴细胞增殖IC₅₀值为药效指标时第1、2、6、7、17、18号峰为药效峰;以IFN-γ分泌抑制率为指标时,第1、2、6、7、8、9、12、13号峰为药效峰。结论 第1、2、6、7、8、9、12、13、17、18号峰所代表的化合物可能为雷公藤多苷片中发挥免疫抑制主要有效成分。

Objective To investigate the immunosuppressive components in Leigongteng Duogan Tablet (LDT). Methods High performance liquid chromatography (HPLC) was applied to establish the fingerprint for LDT from seven manufactures. The immunosuppressive effect of LDT was observed on Con A-induced mice spleen cells, and the cell proliferation and contents of IFN-γ were recorded. Then the partial least squares regression (PLSR) analysis was used to investigate its spectrum-effect relationship. Results The HPLC fingerprint for LDT from seven manufactures was established, and 18 characteristic peaks were confirmed. The in vitro cell experimental results showed that the proliferation of mice spleen cells and secretion of IFN-γ were inhibited by all LDT significantly compared with the control group. According to the results of PLSR, No. 1, 2, 6, 7, 17, and 18 peaks were strongly related to the proliferation inhibition of mice spleen cells and No. 1, 2, 6, 7, 8, 9, 12, and 13 peaks were related to the secretion of IFN-γ. Conclusion The No. 1, 2, 6, 7, 8, 9, 12, 13, 17, and 18 peaks are the principal immunosuppressive substances in LDT.

福建省科技计划重大项目(2012I1001)