目的 穿山甲传统生品不入药，需炮制后方可入药，因此对穿山甲高温砂炒的炮制机制进行研究。方法 采用TLC、Nano LC-Q Exactive Orbitrap MS对穿山甲炮制前后的脂溶性成分及蛋白成分的变化进行分析，同时，对炮制过程中显著增加的环二肽成分的形成进行模拟炮制。并对L-丝-L-酪环二肽的活性进行筛选。结果 穿山甲经砂炒炮制后，脂溶性成分无显著变化、蛋白成分显著减少，环二肽显著增加。环二肽的形成主要与炮制过程中的受热有关，在低温条件下，直链肽N端可环化生成L构型的环二肽，在高温环境下，直链肽N端及C端均可快速环化生成环二肽。L-丝-L-酪环二肽可延长凝血时间并增加奶牛乳腺上皮细胞增殖率和乳蛋白合成相关基因表达，同时还具有显著的镇痛活性，这与穿山甲传统功效相一致。结论 穿山甲经炮制后产生的L-丝-L-酪环二肽可能是穿山甲炮制增效的物质基础。这对揭示穿山甲药效物质基础、寻找穿山甲替代资源、保护穿山甲具有重要意义。

Objective Traditional raw pangolin products are not used as medicine, which can only be used as medicine after processing. Therefore, the processing mechanism of high temperature sand-fried pangolin was studied. Methods The changes of liposolubility and protein composition of pangolin before and after processing were analyzed by TLC and Nano LC-Q Exactive Orbitrap MS. Meanwhile, the simulation processing of cyclic dipeptides, which were significantly increased during processing, was performed. The activity of L-serine-L-tyrosine cyclic dipeptide was screened. Results The results showed that there was no significant change in fat-soluble components, significant decrease in polypeptides and significant increase in cyclic dipeptides after the sand-fried processing of pangolin. The formation of cyclic dipeptides was mainly related to the heating of the processing. At low temperature, the N-terminal of the linear peptide could be cycled to form L-shaped cyclic dipeptides. At high temperature, the N-terminal and C-terminal of the linear peptide could be rapidly cycled to form cyclic dipeptides. L-serine-L-tyrosine cyclic dipeptide could prolong coagulation time and increase the proliferation rate of mammary epithelial cells and the expression of genes related to milk protein synthesis in dairy cows. It also had significant analgesic activity, which was consistent with the traditional efficacy of pangolin. Conclusion These results suggested that large amounts of L-serine-L-tyrosine cyclic dipeptide produced by the processing of pangolin may be one of the material bases for enhancing the processing efficiency of pangolin. It was of great significance for revealing the material basis of pharmacodynamics of pangolin, searching for alternative resources and protecting pangolin.

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苏州市科学技术局民生科技项目"基于酪氨酸环肽同系物的炮山甲通乳物质基础研究"（SYS201787）