基于靶向代谢组学研究苍术麸炒前后干预脾虚大鼠氨基酸水平变化的影响  

Targeted Metabolomic Analysis of Amino Acids with Spleen Deficiency Syndrome Rats after Administering Raw and Bran-Fried Cangzhu(Atractylodis Rhizoma)

作　　者：薛东华[1] 徐宁阳[1] 金珈汐 凌嘉阳 李星宜 崔名杨 胡宇晴 赵煜 鞠宝兆[1] 才谦[1] XUE Donghua;XU Ningyang;JIN Jiaxi;LING Jiayang;LI Xingyi;CUI Mingyang;HU Yuqing;ZHAO Yu;JU Baozhao;CAI Qian(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Eneral Hospital of Northern Theater Command,China Medical University,Shenyang 110003,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]中国人民解放军北部战区总医院,辽宁沈阳110003

出　　处：《中华中医药学刊》2024年第4期113-119,I0025-I0029,共12页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81973478);辽宁省博士启动基金项目(2019-BS-161);辽宁省“兴辽英才计划”项目(XLYC2002004)。

摘　　要：目的研究采用靶向代谢组学方法,观察苍术麸炒前后对脾虚大鼠血清氨基酸水平的变化,探讨苍术炮制机制。方法72只大鼠随机分成正常组、模型组、低剂量生苍术组(生低组)、中剂量生苍术组(生中组)、高剂量生苍术组(生高组)、低剂量麸炒苍术组(麸炒低组)、中剂量麸炒苍术组(麸炒中组)和高剂量麸炒苍术组(麸炒高组),共8组,每组9只。除正常组外,模型组通过过度疲劳、苦寒泻下和饮食不节法造模。对各组大鼠采用酶联免疫吸附测定(Enzyme Linked Immunosorbent Assay,ELISA)方法测定颌下腺中水通道蛋白3(Aquaporin-3,AQP-3)、水通道蛋白4(AQP-4)、水通道蛋白5(AQP-5)、水通道蛋白8(AQP-8)的含量,结肠中水通道蛋白1(AQP-1)、水通道蛋白2(AQP-2)、水通道蛋白9(AQP-9)、紧密连接蛋白1(zonula occludes protein-1,ZO-1)的含量。对生中组和麸炒中组采用超高效液相色谱-三重四极杆质谱(Ultra high performance liquid chromatography-triple quadrupole tandem mass spectrometry,UH-PLC-MS/MS)检测血清中24种氨基酸水平的变化。结果与正常组比较,模型组中颌下腺AQP-3、AQP-4、AQP-5、AQP-8的含量显著降低(P<0.05);模型组中结肠AQP-1和AQP-2的含量显著升高(P<0.05),AQP-9和ZO-1的含量显著降低(P<0.05)。生苍术和麸炒苍术对上述指标均有调节作用。经进一步同等剂量的生苍术和麸炒苍术相比,麸炒中组使上述指标含量升高更明显。与正常组比较,模型组血清中L-缬氨酸等4种物质的水平显著升高(P<0.05),L-酪氨酸等11种物质的水平均显著降低(P<0.05)。经进一步比较,麸炒苍术对上述指标调节效果优于生苍术。靶向代谢通路分析表明,这些差异代谢物主要与精氨酸和脯氨酸代谢、氨酰tRNA生物合成、甘氨酸、丝氨酸和苏氨酸代谢、乙醛酸和二羧酸代谢通路有关。结论苍术炮制后的增效机制可能与对脾虚大鼠血清中氨基酸调节有关。Objective This study used targeted metabonomics to observe the effects of raw and bran-fried crude Cangzhu(Atractylodis Rhizoma)on the changes of serum amino acid levels in rats with spleen deficiency syndrome rats and explore the processing mechanism of Cangzhu(Atractylodis Rhizoma).Methods Seventy-two rats were randomly assigned to control group(CG),model group(MD),raw Cangzhu(Atractylodis Rhizoma)low-dose group(LR),raw Cangzhu(Atractylodis Rhizoma)medium-dose group(MR),raw Cangzhu(Atractylodis Rhizoma)high dose group(HR),bran-fried Cangzhu(Atractylodis Rhizoma)low-dose group(LB),bran-fried Cangzhu(Atractylodis Rhizoma)medium dose group(MB)and bran-fried Cangzhu(Atractylodis Rhizoma)high dose group(HB),eight groups with 9 in each group.In addition to the control group,the model group adopted the complex method of improper diet,excessive fatigue and bitter cold diarrhea to create the spleen deficiency model of rats.The contents of aquaporin(AQP)-3,AQP-4,AQP-5 and AQP-8 in submandibular gland,the contents of AQP-1,AQP-2,AQP-9 and zonula occludes protein-1(ZO-1)in intestinal were determined by enzyme-linked immunosorbent assay(ELISA).The changes of 24 kinds of amino acids in serum of MR and MB groups were detected by Ultra High Performance Liquid Chromatography-Triple Quadrupole Tandem Mass Spectrometry(UHPLC-MS/MS).Results Comparing with those of the CG group,there were lower levels of AQP-3,AQP-4,AQP-5 and AQP-8 in submandibular gland of MG group(P<0.05),higher levels of AQP-1 and AQP in intestinal of MG group(P<0.05),lower levels of AQP-9 and ZO-1in intestinal of MG group(P<0.05).Both raw and bran-fried crude CangZhu(Atractylodis Rhizoma)can regulate the above indexes.Afher further comparison with the same dose of raw and bran-fried crude CangZhu(Atractylodis Rhizoma),the content of the above indexes in the MB group was higher than in the HR group.Compared with those of the CG group,the levels of L-Valine and 4 kinds of substances in the serum increased significantly(P<0.05),while the levels of L-Tyrosine and 1

关 键 词：苍术 靶向代谢组学 氨基酸 炮制 脾虚 

分 类 号：R285.5[医药卫生—中药学]