利用LC-MS/MS技术分析蜂毒对大鼠内源性代谢的影响

为了探究蜂毒对Sprague-Dawley (SD)大鼠机体内源性代谢的影响，采用LC-MS/MS技术分析了蜂毒对大鼠血清中代谢物及可能涉及的代谢通路的影响。选取年龄、性别、身体状况相同的SD大鼠，设置为蛰刺组(A组)、创伤组(B组)和对照组(C组)。对A、B两组大鼠分别进行蜜蜂蛰刺和针蛰刺处理，处理后10 min取血清进行LC-MS/MS分析。正负离子模式下，采用偏最小二乘判别分析(PLS-DA)方法进行数学建模，并验证其稳定性。结果表明，蜇刺组、创伤组、对照组的两两之间受影响代谢物种类及数量均有显著差异，其中71种物质与蜂蛰相关，39种物质与蜂毒相关，24种物质与创伤相关。经进一步筛选，共得到与蜂毒相关的9个比较主要差异代谢物，分别为肉毒碱、溶血磷酯酸、溶血磷脂酰胆碱、乙酰肉毒碱、白介素20、亚油酸、溶血磷酯酰乙醇胺、鞘氨醇、溶血磷脂酰丝氨酸；以及4个受影响代谢通路，包括亚油酸代谢、神经鞘脂类代谢、甘油磷脂代谢、丙氨酸和天门冬氨酸代谢。蛰刺、创伤和蜂毒均对大鼠的内源性代谢产生一定影响，其中，与创伤相比，蜂毒影响相对更大。根据受影响代谢通路的生理作用，推测蜂毒对机体内源性代谢的影响主要涉及脂肪酸的代谢过程，具有重要的生理学作用。

Influence of Bee Venom on Endogenous Metabolism of Sprague-Dawley Rats Using LC-MS/MS

To explore the influence of bee venom on endogenous metabolism, this study used liquid chromatography-mass spectrometry (LC-MS/MS) to analyze the effect of bee venom on the metabolites and possibly involved metabolism pathways in rats serum. Sprague Dawley (SD) rats with the same age, the same gender and the same physical condition were chosen and divided into 3 groups including bee sting group (group A), stabbing group (group B) and the control group (group C) at random. Rats in group A and B were single stung by honeybees and needle, respectively. 10 min after sting, serum specimens from these 3 groups would be analyzed by LC-MS/MS. In positive and negative ion mode, partial least squares-discriminate analysis was adopted to establish mathematical model, and their stability would be tested later. The results indicated that there were significant differences in types and quantities of influenced metabolites among different groups. The number of differential metabolite related to bee venom, sting and stabbing were 71, 39 and 24, respectively. After further screening, 9 potential biomarkers were found showing relation with bee venom, including carnitine, acetylcarnitine, Lyso PC, Lyso-PA, lyso-PS, lyso-PE, IL-20, linoleic, and sphinganine. And they were mainly involved in 4 metabolic pathways, including linoleic acid metabolism, sphingolipids catabolism, phospholipid catabolism, Alanine and Aspartate anabolism. The results indicated that sting, stabbing and bee venom all influence endogenous metabolites of SD rats. Comparing with stabbing, bee venom showed relatively stronger effect. According to the physiological function of affected metabolic pathways, it was speculated that the effect of bee venom on endogenous metabolisms was mainly involved in the metabolic process of fatty-acid, and exploration about this process had significant meaning for physiology.