[关键词]
[摘要]
目的 评价高良姜素(GA)对斑马鱼整体形态及心、肝、肾主要脏器的影响。方法 采用不同浓度(1、2、4、6、8、12、18、28、30、32、64 μmol·L-1)高良姜素处理受精后48 h(48 hpf)的斑马鱼,给药24、48 h后(24、48 hpe),计算斑马鱼死亡率和畸形率;显微镜下观察高良姜素对斑马鱼形态、心率、心包面积、静脉窦-动脉球(SV-BA)距离、肝脏、卵黄囊吸收及眼部水肿情况的影响。结果 斑马鱼暴露在1、2、4、6、8、12 μmol·L-1浓度下的高良姜素均无死亡;24 hpe,高良姜素对斑马鱼的半数致死浓度(LC50)为30 μmol·L-1;48 hpe的LC50为28 μmol·L-1。与空白对照组比较,24 hpe ,浓度≥ 8 μmol·L-1的高良姜素处理斑马鱼畸形率显著上升(P<0.05);48 hpe,浓度≥6 μmol·L-1的高良姜素处理斑马鱼畸形率显著升高(P<0.01)。与空白对照组相比,4 μmol·L-1及以下浓度的高良姜素对斑马鱼形态及心、肝、肾无明显影响。与空白对照组相比,在24 hpe,8 μmol·L-1高良姜素组斑马鱼发育评分、相对体长显著降低(P<0.001),心脏出现轻微的心包水肿、心包面积显著增加(P<0.05)、心率及SV-BA距离无明显变化,肝脏面积和肝脏荧光强度显著减小(P< 0.001),卵黄囊面积显著增加(P<0.001),肾脏无明显变化;在48 hpe,6 μmol·L-1浓度及以上高良姜素组发育评分显著降低(P<0.05、0.001),斑马鱼心脏出现轻微的心包水肿、心包面积显著增加(P<0.001)、心率显著减小(P<0.01)、SV-BA距离显著增加(P< 0.01),肝脏面积和肝脏荧光强度显著减小(P<0.01、0.001),卵黄囊面积显著增加(P<0.01、0.001),8 μmol·L-1组相对体长显著降低(P<0.001)。结论 高良姜素在4 μmol·L-1及以下对斑马鱼无毒性影响;6 μmol·L-1及以上浓度对斑马鱼有轻微的心脏毒性和肝脏毒性,在临床应用中要合理控制用量。
[Key word]
[Abstract]
Objective To evaluate the effects of Galangin (GA) on the overall morphology and major organs of heart, liver and kidney of zebrafish. Methods The zebrafish 48 h (48 hpf) after fertilization were treated with GA at different concentrations (1, 2, 4, 6, 8, 12, 18, 28, 30, 32, 64 μmol·L-1). After 24 and 48 hours of experimental treatment (24 and 48 hpe), the mortality and malformation rates of zebrafish were calculated. The effects of GA on the overall morphology, heart rate, pericardial area, SV-BA distance, liver and yolk sac absorption and eye edema of zebrafish were observed. Results Zebrafish exposed to a concentration of 1, 2, 4, 6, 8, 12 μmol·L-1, there was no death of galangin. 24 hpe, the LC50 of galangin to zebrafish was 30 μmol·L-1. The LC50 for 48 hpe was 28 μmol·L-1. Compared with the blank control group, 24 hpe, the deformity rate of zebrafish significantly increased with concentration ≥ 8 μmol·L-1 (P< 0.05), 48 hpe the deformity rate of zebrafish was significantly increased (P< 0.01) when treated with high quality curcumin of 6 μmol·L-1. Compared with the blank control group, the concentration of galangin at or below 4 μmol·L-1 had no significant effect on the morphology, heart, liver, and kidney of zebrafish. Compared with the blank control group, at 24 hpe, in the 6 μmol·L-1 galangin group, the development score and relative body length of zebrafish were significantly reduced (P< 0.001), with mild pericardial edema in the heart, significant increase in pericardial area (P< 0.05), no significant changes in heart rate and SV-BA distance, significant decrease in liver area and fluorescence intensity (P< 0.001), significant increase in yolk sac area (P< 0.001), and no significant changes in the kidneys. At 48 hpe, in the ≥ 8 μmol·L-1 galangin group, the development score significantly decreased (P< 0.05, 0.001), and the heart of zebrafish showed slight pericardial edema, significant increase in pericardial area (P< 0.001), significant decrease in heart rate (P< 0.01), significant increase in SV-BA distance (P< 0.01), significant decrease in liver area and liver fluorescence intensity (P< 0.01, 0.001), and significant increase in yolk sac area (P< 0.01, 0.001). The relative body length of the 8 μmol·L-1 group was significantly reduced (P< 0.001). Conclusion GA showed no toxicity on zebrafish at 4 μmol·L-1 and below. The concentration of GA above 6 μmol·L-1 has slight cardiotoxicity and hepatotoxicity to zebrafish, so it is necessary to control the dosage reasonably in clinical application.
[中图分类号]
R965.3
[基金项目]
自治区自然科学基金项目(2021D01A156);国家自然科学地区科学基金项目(82160821);天山英才项目(2022TSYCCX0021,2022TSYCLJ009);自治区重点研发项目(2022B03012-4);2020年新疆维吾尔自治区高层次人才引进工程(柔性引进人才)项目资助