CAJ | 학술논문

目的：通过对脑泰方含药血清和含药血浆蛋白质（肽）组学的比较研究，发现脑泰方含药血清和含药血浆中的差异蛋白（肽）；以质谱分析技术确定差异蛋白的特征。从而为探索脑泰方的药效物质基础和发展中药血浆药理学方法提供科学依据。方法：取健康成年SD大鼠20只，按体重随机分为空白组、脑泰方给药组，每组10只，连续灌胃给药7天。手术前12 h禁食不禁水，末次给药后1 h，用10%水合氯醛腹腔麻醉，经颈总动脉插管取血。每只大鼠采血后分别制备血清和血浆。每组随机抽取5只大鼠的血清及血浆，采用2-DE技术进行蛋白质（肽）组学比较研究,运用300DPI扫描、PDQuest 7.3.0软件进行分析；采用ESI-MS/MS对重要差异蛋白质点进行鉴定，筛选脑泰方血清和血浆差异蛋白的特征。结果：共获得20张胶图，各胶图分别得到664个蛋白位点。其中与空白血浆组比较，脑泰方血浆样品中共识别到20个差异蛋白点，其中15个差异点表达上调，5个差异点表达下调；与空白血清组比较，脑泰方血清样品中共识别到19个差异蛋白点，其中15个差异点表达上调，4个差异点表达下调；与脑泰方血浆组比较，脑泰方血清样品中共识别到24个差异蛋白点，其中9个差异点表达上调，15个差异点表达下调。选取在脑泰方血浆组和脑泰方血清组中高表达的10个差异蛋白质点进行基质辅助激光解析电离飞行时间质谱（MAL-DI-TOF-MS），成功鉴定了6个蛋白，分别是：inter-alpha trypsin inhibitor, heavy chain 3、Group specific component、complement factor B、Receptor Complexed With A Heterodimeric Fc、complement factor B, isoform CRA-d、Transferrin。结论：脑泰方含药血浆、血清中明显变化的蛋白质可能与其抗凝血促纤溶作用机制相关，这些蛋白参与血管新生、炎症调控等病理生理过程，对研究脑泰方作用的可能有效靶点及其信号转导通路意义重大。目的：通過對腦泰方含藥血清和含藥血漿蛋白質（肽）組學的比較研究，髮現腦泰方含藥血清和含藥血漿中的差異蛋白（肽）；以質譜分析技術確定差異蛋白的特徵。從而為探索腦泰方的藥效物質基礎和髮展中藥血漿藥理學方法提供科學依據。方法：取健康成年SD大鼠20隻，按體重隨機分為空白組、腦泰方給藥組，每組10隻，連續灌胃給藥7天。手術前12 h禁食不禁水，末次給藥後1 h，用10%水閤氯醛腹腔痳醉，經頸總動脈插管取血。每隻大鼠採血後分彆製備血清和血漿。每組隨機抽取5隻大鼠的血清及血漿，採用2-DE技術進行蛋白質（肽）組學比較研究,運用300DPI掃描、PDQuest 7.3.0軟件進行分析；採用ESI-MS/MS對重要差異蛋白質點進行鑒定，篩選腦泰方血清和血漿差異蛋白的特徵。結果：共穫得20張膠圖，各膠圖分彆得到664箇蛋白位點。其中與空白血漿組比較，腦泰方血漿樣品中共識彆到20箇差異蛋白點，其中15箇差異點錶達上調，5箇差異點錶達下調；與空白血清組比較，腦泰方血清樣品中共識彆到19箇差異蛋白點，其中15箇差異點錶達上調，4箇差異點錶達下調；與腦泰方血漿組比較，腦泰方血清樣品中共識彆到24箇差異蛋白點，其中9箇差異點錶達上調，15箇差異點錶達下調。選取在腦泰方血漿組和腦泰方血清組中高錶達的10箇差異蛋白質點進行基質輔助激光解析電離飛行時間質譜（MAL-DI-TOF-MS），成功鑒定瞭6箇蛋白，分彆是：inter-alpha trypsin inhibitor, heavy chain 3、Group specific component、complement factor B、Receptor Complexed With A Heterodimeric Fc、complement factor B, isoform CRA-d、Transferrin。結論：腦泰方含藥血漿、血清中明顯變化的蛋白質可能與其抗凝血促纖溶作用機製相關，這些蛋白參與血管新生、炎癥調控等病理生理過程，對研究腦泰方作用的可能有效靶點及其信號轉導通路意義重大。
목적：통과대뇌태방함약혈청화함약혈장단백질（태）조학적비교연구，발현뇌태방함약혈청화함약혈장중적차이단백（태）；이질보분석기술학정차이단백적특정。종이위탐색뇌태방적약효물질기출화발전중약혈장약이학방법제공과학의거。방법：취건강성년SD대서20지，안체중수궤분위공백조、뇌태방급약조，매조10지，련속관위급약7천。수술전12 h금식불금수，말차급약후1 h，용10%수합록철복강마취，경경총동맥삽관취혈。매지대서채혈후분별제비혈청화혈장。매조수궤추취5지대서적혈청급혈장，채용2-DE기술진행단백질（태）조학비교연구,운용300DPI소묘、PDQuest 7.3.0연건진행분석；채용ESI-MS/MS대중요차이단백질점진행감정，사선뇌태방혈청화혈장차이단백적특정。결과：공획득20장효도，각효도분별득도664개단백위점。기중여공백혈장조비교，뇌태방혈장양품중공식별도20개차이단백점，기중15개차이점표체상조，5개차이점표체하조；여공백혈청조비교，뇌태방혈청양품중공식별도19개차이단백점，기중15개차이점표체상조，4개차이점표체하조；여뇌태방혈장조비교，뇌태방혈청양품중공식별도24개차이단백점，기중9개차이점표체상조，15개차이점표체하조。선취재뇌태방혈장조화뇌태방혈청조중고표체적10개차이단백질점진행기질보조격광해석전리비행시간질보（MAL-DI-TOF-MS），성공감정료6개단백，분별시：inter-alpha trypsin inhibitor, heavy chain 3、Group specific component、complement factor B、Receptor Complexed With A Heterodimeric Fc、complement factor B, isoform CRA-d、Transferrin。결론：뇌태방함약혈장、혈청중명현변화적단백질가능여기항응혈촉섬용작용궤제상관，저사단백삼여혈관신생、염증조공등병리생리과정，대연구뇌태방작용적가능유효파점급기신호전도통로의의중대。
Through comparative study on Naotaifang containing serum and plasma proteomics (peptide), this article revealed differential proteins (peptides) in the Naotaifang. The characteristics of differential proteins were identified with mass spectrometry. It provides scientific evidences for the pharmacodynamic material basis and Chinese herbal medicine plasma pharmacological method development in the exploration of Naotaifang. A total of 20 healthy adult SD rats were randomly divided into the control group, Naotaifang treatment group according to their weights. Ten rats in each group. Intragastric administration of medication was given for seven consecutive days. Before surgery, rats were fed with water but without food. One hour after the last drug administration, 10% chloral hydrate was injected for intraperitoneal anesthesia. Blood was taken through the common carotid artery. Serum and plasma samples were made after blood was taken from each rat. Serum and plasma samples of five rats were randomly selected from each group. And the two-dimensional electrophoresis (2-DE) technique was used in the comparative study of serum pro-teomics (peptide). The 300 DPI scanning and PDQuest 7.3.0 were used in the analysis. The ESI-MS/MS was used to identify important differences in proteins and screen characteristic serum and plasma protein. The results showed that 20 differential proteins of 5 plasma samples were identified. There were 15 types of proteins expressing up-regulation and 5 types expressing down-regulation. Comparative analysis on the 2-DE gel pictures of Naotaifang containing serum, 19 differential proteins of 5 plasma samples were identified, among which 15 types of proteins express up-regulation and 4 down-regulation. Comparative analysis on the 2-DE gel pictures of Naotaifang containing serum and Naotaifang containing plasma showed that 24 differential proteins of 5 plasma samples were identified, among which 9 types of proteins express up-regulation and 15 down-regulation. The highly expressed proteins were selected to MALDI-TOF-MS between Naotaifang containing serum and Naotaifang containing plasma. There were six successful-ly identified proteins, which were inter-alpha trypsin inhibitor, heavy chain 3, group-specific component, comple-ment factor B, Receptor Complexed with A Heterodimeric Fc, isoform CRA-d, Transferrin. It was concluded that protein with obvious changes in the Naotaifang containing serum and plasma may be related with fibrinolysis and an-ticoagulant. These proteins are involved in angiogenesis, inflammation and other pathological regulations of physiolog-ical processes. They are of great significance in the study of effective target and its signal transduction pathway of Naotaifang.