目的 應(yīng)用斑馬魚(yú)模型評(píng)價(jià)注射用益氣復(fù)脈（凍干）（YQFM）的心血管毒性和肝毒性。方法 受精后48 h的野生型AB系斑馬魚(yú)隨機(jī)分為空白對(duì)照組（未注射組）、溶劑對(duì)照組（注射0.9%氯化鈉注射液10 nL）和YQFM 10、20、100、200、400、600、800和1 000 ng組，iv給藥4 h后統(tǒng)計(jì)各劑量組斑馬魚(yú)死亡數(shù)量，計(jì)算最大非致死劑量（MNLD）和10%致死劑量（LD₁₀）值。心血管毒性和肝毒性評(píng)價(jià)劑量設(shè)置為MNLD/10（20 ng）、MNLD/3（60 ng）、MNLD（200 ng）和LD₁₀（400 ng）；在顯微鏡下觀察YQFM對(duì)斑馬魚(yú)心率的影響，觀察斑馬魚(yú)有無(wú)心包水腫、血流速度變化、血液流失、心包瘀（出）血、血栓等，計(jì)算各表型發(fā)生率；在顯微鏡下對(duì)肝臟進(jìn)行觀察，對(duì)典型表型進(jìn)行拍照，利用分析軟件NIS-ElementsD 3.10分析肝臟面積、肝臟不透光度和卵黃囊面積。結(jié)果 YQFM MNLD和LD₁₀分別為198.06 ng（約為200 ng）和397.12 ng（為400 ng）；與溶劑對(duì)照組比較，YQFM 20、60 ng組斑馬魚(yú)未見(jiàn)異常，沒(méi)有觀察到心血管毒性，未見(jiàn)心包水腫，未見(jiàn)心率減慢，未見(jiàn)心律不齊；YQFM 200 ng組斑馬魚(yú)出現(xiàn)發(fā)生率較低的心血管毒性，表現(xiàn)為心包水腫（發(fā)生率僅為3.3%），未見(jiàn)心率減慢，未見(jiàn)心律不齊；400 ng組斑馬魚(yú)出現(xiàn)較輕微的心血管毒性，表現(xiàn)為心包水腫（發(fā)生率為13.3%）、血流變慢（發(fā)生率為6.7%）、血流缺失（發(fā)生率3.3%），未見(jiàn)心率減慢，未見(jiàn)心律不齊。與溶劑對(duì)照組比較，YQFM各劑量（20、60、200、400 ng）組斑馬魚(yú)相對(duì)肝臟大小、肝臟不透光度平均值和卵黃囊延遲吸收倍數(shù)均無(wú)統(tǒng)計(jì)學(xué)差異。結(jié)論 YQFM在MNLD劑量下，不誘發(fā)斑馬魚(yú)心血管毒性和肝臟毒性。

Objective The cardiotoxicity and hepatotoxicity of Yiqi Fumai Lyophilized Injection (YQFM) were evaluated by zebrafish model in vivo experiment. Methods At 48 h after fertilization, wild type AB zebrafish were randomly divided into blank control group (no injection group), solvent control group (injection of 0.9% sodium chloride injection 10 nL) and YQFM 10, 20, 100, 200, 400, 600, 800 and 1 000 ng groups. 4 h after iv administration, the death number of zebrafish in each dose group was counted, and the maximum non-lethal dose (MNLD) and 10% lethal dose (LD₁₀) values were calculated. The evaluation doses of cardiovascular toxicity and hepatotoxicity were set as MNLD/10 (20 ng), MNLD/3 (60 ng), MNLD (200 ng) and LD₁₀ (400 ng). The effects of YQFM on the heart rate of zebrafish were observed under a microscope, and the incidence of each phenotype was calculated. The liver was observed under a microscope and the typical phenotypes were photographed. The liver area, liver impermeability and yolk sac area were analyzed using nIS-Elements D 3.10 software. Results The MNLD and LD₁₀ of YQFM were 198.06 ng (about 200 ng) and 397.12 ng (400 ng), respectively. Compared with solvent control group, zebrafish in YQFM 20 and 60 ng groups showed no abnormalities, no cardiovascular toxicity, no pericardium edema, no slow heart rate, and no arrhythmia. 200 ng zebrafish showed low incidence of cardiovascular toxicity, such as pericardium edema (3.3%), no slow heart rate, no arrhythmia. In the 400 ng group, zebrafish showed mild cardiovascular toxicity, including pericardium edema (13.3%), hemorheology slowness (6.7%), loss of blood flow (3.3%), and no slow heart rate or arrhythmia. Compared with solvent control group, there were no significant differences in relative liver size, mean value of liver impermeability and delayed absorption ratio of yolk capsule of zebrafish in YQFM (20, 60, 200 and 400 ng) groups. Conclusion YQFM did not induce cardiotoxicity and hepatotoxicity in zebrafish at the MNLD dose.

R965.3

天津市科技計(jì)劃項(xiàng)目（18YFCZZC00430）