Liu Yi, Guo Jing, Zhang June, Deng Yunyun, Xiong Guanghua, Fu Jianpin, Wei Lili, Lu Huiqiang
College of Life Sciences, Jiangxi Normal University, Nanchang, Jiangxi, China.
Jiangxi Engineering Laboratory of Zebrafish Modeling and Drug Screening for Human Diseases; Jiangxi Key Laboratory of Developmental Biology of Organs; College of Life Sciences, Jinggangshan University, Jian, Jiangxi, China.
Aquat Toxicol. 2022 Jan;242:106039. doi: 10.1016/j.aquatox.2021.106039. Epub 2021 Nov 21.
Chlorogenic acid (CGA) is a phenylpropanoid compound that is well known to improve the antioxidant capacity and other biological activities. However, the roles of CGA in the liver development of organisms are unclear. In the present study, we aimed to investigate the function of CGA in the hepatic development in thioacetamide (TAA)-induced zebrafish embryos. We found that CGA exerted certain beneficial effects on zebrafish larvae from TAA-exposed zebrafish embryos, such as increasing the liver size, body length, heart rate, acetylcholinesterase activity, and motor ability. In addition, CGA displayed an antioxidant effect on TAA-induced zebrafish embryos by enhancing the activities of superoxide dismutase (SOD), catalase (CAT), and glucose-6-phosphate dehydrogenase (G6PDH), and decreasing of the contents of malondialdehyde (MDA), reactive oxygen species (ROS), and nitric oxide (NO). The results of western blotting analysis showed that CGA inhibited cell apoptosis by increasing the levels of Bcl2 apoptosis regulator and decreasing the levels of Bcl2 associated X (Bax), apoptosis regulator and tumor protein P53. Moreover, CGA promoted cell proliferation in TAA-induced zebrafish larvae, as detected using proliferating cell nuclear antigen fluorescence immunostaining. In addition, CGA inhibited the expression of Wnt signaling pathway genes Dkk1 (encoding Dickkopf Wnt signaling pathway inhibitors), and promoted the expression of Lef1 (encoding lymphoid enhancer binding factor 1) and Wnt2bb (encoding wingless-type MMTV integration site family, member 2Bb). When the Wnt signal inhibitor IWR-1 was added, there was no significant change in liver development in the IWR-1 + TAA group compared with the IWR-1 + TAA + CGA group (p <0.05), which suggested that CGA regulates liver development via Wnt signaling pathway. Overall, our results suggested that CGA might alleviate TAA-induced toxicity in zebrafish and promote liver development through the Wnt signaling pathway, which provides a basis for the therapeutic effect of CGA on liver dysplasia.
绿原酸（CGA）是一种苯丙烷类化合物，众所周知它能提高抗氧化能力和其他生物活性。然而，CGA在生物体肝脏发育中的作用尚不清楚。在本研究中，我们旨在研究CGA在硫代乙酰胺（TAA）诱导的斑马鱼胚胎肝脏发育中的功能。我们发现，CGA对TAA暴露的斑马鱼胚胎的幼虫有一定的有益作用，如增加肝脏大小、体长、心率、乙酰胆碱酯酶活性和运动能力。此外，CGA通过增强超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和葡萄糖-6-磷酸脱氢酶（G6PDH）的活性，以及降低丙二醛（MDA）、活性氧（ROS）和一氧化氮（NO）的含量，对TAA诱导的斑马鱼胚胎表现出抗氧化作用。蛋白质免疫印迹分析结果表明，CGA通过增加Bcl2凋亡调节因子的水平和降低Bcl2相关X蛋白（Bax）、凋亡调节因子和肿瘤蛋白P53的水平来抑制细胞凋亡。此外，使用增殖细胞核抗原荧光免疫染色检测发现，CGA促进了TAA诱导的斑马鱼幼虫的细胞增殖。此外，CGA抑制Wnt信号通路基因Dkk1（编码Dickkopf Wnt信号通路抑制剂）的表达，并促进Lef1（编码淋巴增强子结合因子1）和Wnt2bb（编码无翅型MMTV整合位点家族成员2Bb）的表达。当加入Wnt信号抑制剂IWR-1时，IWR-1 + TAA组的肝脏发育与IWR-1 + TAA + CGA组相比无显著变化（p <0.05），这表明CGA通过Wnt信号通路调节肝脏发育。总体而言，我们的结果表明，CGA可能减轻TAA诱导的斑马鱼毒性，并通过Wnt信号通路促进肝脏发育，这为CGA对肝脏发育异常的治疗作用提供了依据。
