目的 基于斑馬魚模型探究暈痛定膠囊的鎮(zhèn)痛活性,采用網(wǎng)絡(luò)藥理學(xué)方法預(yù)測(cè)暈痛定膠囊的鎮(zhèn)痛作用機(jī)制。方法 采用佛波酯構(gòu)建斑馬魚疼痛模型,以阿司匹林為陽性對(duì)照,利用行為軌跡分析系統(tǒng)監(jiān)測(cè)斑馬魚的運(yùn)動(dòng)軌跡,以運(yùn)動(dòng)次數(shù)、運(yùn)動(dòng)時(shí)間、運(yùn)動(dòng)距離、運(yùn)動(dòng)速度等運(yùn)動(dòng)狀態(tài)為指標(biāo)評(píng)價(jià)暈痛定膠囊及其組方藥材的鎮(zhèn)痛活性;同時(shí)運(yùn)用網(wǎng)絡(luò)藥理學(xué)及分子對(duì)接技術(shù)預(yù)測(cè)暈痛定膠囊鎮(zhèn)痛核心成分及潛在靶點(diǎn);選取網(wǎng)絡(luò)藥理學(xué)預(yù)測(cè)的核心成分法卡林二醇和金色酰胺醇酯為實(shí)驗(yàn)藥物,利用斑馬魚疼痛模型驗(yàn)證鎮(zhèn)痛活性。結(jié)果 斑馬魚疼痛模型鎮(zhèn)痛活性評(píng)價(jià)實(shí)驗(yàn)中,暈痛定膠囊及其組方藥材川芎、蜜環(huán)菌均能顯著降低模型斑馬魚的快速運(yùn)動(dòng)和慢速運(yùn)動(dòng)(P＜0.01),且暈痛定膠囊作用強(qiáng)于川芎和蜜環(huán)菌。網(wǎng)絡(luò)藥理學(xué)篩選得到暈痛定膠囊鎮(zhèn)痛活性成分87個(gè)及其作用靶點(diǎn)159個(gè),利用蛋白質(zhì)-蛋白質(zhì)相互作用(PPI)網(wǎng)絡(luò)分析得到法卡林二醇和金色酰胺醇酯等核心成分和MAPK1等核心靶點(diǎn),分子對(duì)接結(jié)果顯示核心成分及核心靶點(diǎn)間具有較好的結(jié)合活性。法卡林二醇和金色酰胺醇酯鎮(zhèn)痛活性實(shí)驗(yàn)結(jié)果表明單獨(dú)給藥法卡林二醇和金色酰胺醇酯可以達(dá)到良好的鎮(zhèn)痛效果,與網(wǎng)絡(luò)藥理學(xué)預(yù)測(cè)的核心成分結(jié)果相符。結(jié)論 暈痛定膠囊能夠緩解暴露于佛波酯中斑馬魚的疼痛狀態(tài),具有較明顯的鎮(zhèn)痛作用;預(yù)測(cè)其作用機(jī)制可能與減少神經(jīng)損傷及其炎癥反應(yīng)和促進(jìn)細(xì)胞凋亡等過程有關(guān);且法卡林二醇和金色酰胺醇酯為其發(fā)揮鎮(zhèn)痛作用的核心成分。

Objective To explore the analgesic activity of Yuntongding Capsules(YTD) based on zebrafish model and analgesic mechanism of Yuntongding Capsules by network pharmacology.Methods Phorbol myristate acetate(PMA) was used to establish a zebrafic pain model, and aspirin was used as positive control. The movement trajectory of zebrafish was monitored with the behavior trajectory analysis system and the analgesic activity of YTD was evaluated by the movement states such as movement frequency,movement time, movement distance and movement speed. Network pharmacology and molecular docking techniques were used to predict the core components and potential analgesic mechanism of YTD. Selecting network pharmacology prediction components of falcarindiol(Fal) and aurantiamide acetate(Aur) as the experimental drugs to verify the network pharmacology prediction results.Results By comparing the exercise intensity and trajectory of zebrafish in the model group and each administration groups, YTD has better analgesic activity, and it is better than the analgesic effect of Ligusticum or Armillaria mellea alone. 87 active components and 159 action targets of YTD analgesia were screened by network pharmacology. Core components such as Aur and core targets such as MAPK1 were obtained by protein-protein interaction network analysis and molecular docking was performed, showing that the core component targets had good binding activity. Data from the Fal and Aur groups suggest that favorable analgesia can be achieved by administering Fal or Aur alone. This is consistent with the results of network pharmacology.Conclusion YTD can alleviate the pain of zebrafish exposed to PMA and has obvious analgesic effect. Predicting its mechanism of action may be related to processes such as reducing nerve damage and inflammatory response, and promoting cell apoptosis. At the same time, Fal and Aur are the core components that exert analgesic effects, providing a foundation for the research and development of new analgesic drugs in the future.

R285.5

北京中醫(yī)藥大學(xué)新教師啟動(dòng)基金資助項(xiàng)目(2020-JYB-XJSJJ-009)