目的 通過超高效液相色譜-四級(jí)桿/軌道肼高分辨質(zhì)譜（UPLC-Q-Qrbitrap HRMS）技術(shù)和整合網(wǎng)絡(luò)藥理學(xué)，探究苗藥芪膠升白膠囊治療白細(xì)胞減少癥的作用機(jī)制。方法 將18只大鼠隨機(jī)數(shù)字表法分為空白組和實(shí)驗(yàn)組，每組9只。空白組ig蒸餾水1 mL，實(shí)驗(yàn)組ig芪膠升白膠囊混懸液1 mL（取芪膠升白膠囊的內(nèi)容物，用蒸餾水超聲溶解成混懸液），劑量為2.0 g/kg，連續(xù)給藥3 d，末次給藥劑量為4.0 g/kg。末次給藥后，用1%的戊巴比妥鈉40 mg/kg麻醉大鼠，于15、30、60、120、240 min從大鼠眼眶靜脈叢和尾靜脈交替取血，采集含藥血清和空白血清；運(yùn)用UPLC-Q-Qrbitrap HRMS技術(shù)定性檢測(cè)芪膠升白膠囊膠囊成分、含藥血清成分和空白血清成分，比對(duì)分析得到芪膠升白膠囊的入血成分，再對(duì)入血成分構(gòu)建網(wǎng)絡(luò)藥理學(xué)分析模型，通過成分拓?fù)浞治觥⒌鞍紫嗷プ饔镁W(wǎng)絡(luò)（PPI）分析和通路富集分析，得到芪膠升白膠囊的關(guān)鍵成分、關(guān)鍵靶點(diǎn)和富集通路。結(jié)果 檢測(cè)出芪膠升白膠囊中的成分51種，包括淫羊藿苷、苦參堿、槲皮素等；入血成分58種，其中原型入血成分17種，代謝成分41種，包括金雀異黃酮、槲皮素、5,6,7-三甲氧基香豆素、碳環(huán)素、脫水淫羊藿素、亞油酸、咖啡酸等；網(wǎng)絡(luò)藥理學(xué)分析得到入血成分的靶點(diǎn)基因959個(gè)、白細(xì)胞減少癥的靶點(diǎn)基因3 346個(gè)，交集靶點(diǎn)404個(gè)， PPI分析預(yù)測(cè)出蛋白酪氨酸性磷酸非受體型11（PTPN11）、淋巴細(xì)胞特異蛋白酪氨酸激酶（LCK）、信號(hào)轉(zhuǎn)導(dǎo)及轉(zhuǎn)錄激活蛋白3（STAT3）、磷脂酰肌醇-3-激酶催化亞基α（PIK3CA）、磷脂酰肌醇3-激酶調(diào)節(jié)亞基1（PIK3R1）等核心靶點(diǎn)，通路富集分析發(fā)現(xiàn)芪膠升白膠囊可以通過磷脂酰肌醇-3-羥激酶（PI3K）-蛋白激酶B（Akt）、脂質(zhì)與動(dòng)脈粥樣硬化、Janus激酶/信號(hào)轉(zhuǎn)導(dǎo)與轉(zhuǎn)錄激活子（JAK-STAT）、叉頭框蛋白O（FoxO）、造血細(xì)胞譜系、亞油酸代謝等信號(hào)通路防治白細(xì)胞減少癥。結(jié)論 明確了芪膠升白膠囊的膠囊成分和入血成分，探究了芪膠升白膠囊可通過多個(gè)核心靶點(diǎn)作用于PI3K-AKT、脂質(zhì)與動(dòng)脈粥樣硬化、JAK-STAT等多條通路來治療白細(xì)胞減少癥，為進(jìn)一步研究苗藥芪膠升白膠囊的物質(zhì)基礎(chǔ)和深入闡明其作用機(jī)制提供了理論依據(jù)。

Objective To explore the material basis and mechanism of the Qijiao Shengbai Capsules in treatment of leukopenia by using UPLC-Q-Qrbitrap HRMS and network pharmacology. Methods A total of 18 rats were randomly divided into blank group and experimental group with 9 rats in each group. The blank group was given 1 mL distilled water intragastrically, and the experimental group received 1 mL of Qijiao Shengbai Capsules suspension intragastrically (the contents of Qijiao Shengbai Capsules were taken and ultrasonically dissolved into suspension with distilled water), the dose was 2.0 g/kg, and the dose was 4.0 g/kg for 3 consecutive days. After the last administration, the rats were anesthetized with 1% pentobarbital sodium 40 mg/kg. At 15, 30, 60, 120, and 240 min, blood was taken alternately from the orbital venous plexus and the caudal vein. The drug-containing serum and blank serum were collected, and the UPLC-Q-Qrbitrap HRMS technology was used to qualitatively detect the Qijiao Shengbai Capsules components, drug-containing serum components, and blank serum components of Qijiao Shengbai Capsules. The blood entry components of Qijiao Shengbai Capsules were obtained by comparative analysis, and the network pharmacological analysis model was constructed for the blood entry components. Through component topology analysis, protein interaction network (PPI) analysis and pathway enrichment analysis, the key components, key targets and enrichment pathway of Qijiao Shengbai Capsules were obtained. Results 51 Main components of Qijiao Shengbai Capsules were identified, including icariin, matrine, quercetin, etc. There were 58 blood components, including 17 prototypical blood components and 41 metabolic components, including genistein, quercetin, 5,6, 7-trimethoxycoumarin, carbaprostacyclin, icaritin, linoleic acid, caffeic acid , etc. Network pharmacological analysis revealed 959 target genes for blood entry components, 3 346 target genes for leukopenia, and 404 intersection targets. PPI analysis predicted the core targets of PTPN11, LCK, STAT3, PIK3CA, PIK3R1, etc. Pathway enrichment analysis showed that Qijiao Shengbai Capsules could be regulated by PI3K-Akt, lipid and atherosclerosis, JAK-STAT, FoxO, hematopoietic cell lineage, and linoleic acid metabolism and other signaling pathways to prevent leukopenia. Conclusion The capsule composition and blood entry composition of Qijiao Shengbai Capsules were clarified, and it was explored that Qijiao Shengbai Capsules could treat leukopenia by acting on PI3K-Akt, lipid and atherosclerosis, JAK-STAT and other pathways through multiple core targets, which provided a theoretical basis for further research on the material basis of Qijiao Shengbai Capsules and further elucidate its mechanism of action.

R285

國家重點(diǎn)研發(fā)計(jì)劃項(xiàng)目（2020YFC2005005）