Lu Jianing, Lu Jing, Bu Xiujuan, Li Yazhuo, Ge Guangcai, Guan Shuang
College of Food Science and Engineering, Jilin University, Changchun, Jilin, People's Republic of China.
Key Laboratory of Zoonosis, Ministry of Education College of Veterinary Medicine, Jilin University, Changchun, Jilin, People's Republic of China.
J Food Sci. 2021 Dec;86(12):5503-5515. doi: 10.1111/1750-3841.15968. Epub 2021 Nov 23.
In recent years, foodborne pollutants have become a hot issue in the field of food safety. 3-chloro-1,2-propanediol (3-MCPD) is a widely existing food contaminant. In our previous study, it was confirmed that 3-MCPD can block autophagic flux by inhibiting lysosomal function, thus causing liver injury. Ginseng is a traditional Chinese herbal medicine that contains a variety of bioactive ingredients, among which ginsenoside Rb1 (Gs-Rb1) is the most abundant. In this study, we aim to use Gs-Rb1 to improve 3-MCPD-induced autophagic flux blockage to alleviate liver injury. First, a nontoxic dose of Gs-Rb1 was identified by screening with the MTT method in which Gs-Rb1was added to HepG2 cells and co-treated with 3-MCPD. We found that Gs-Rb1 effectively enhanced the cell activity inhibited by 3-MCPD. Meanwhile, apoptosis data showed that Gs-Rb1 significantly alleviated the apoptosis of HepG2 cells induced by 3-MCPD. Subsequently, we found that Gs-Rb1 could alleviate autophagic flux blockage caused by 3-MCPD in a dose-dependent manner by detecting autophagy-related protein levels and transfecting mRFP-GFP-LC3 adenovirus. On this basis, we used Western blotting and qPCR to explore whether miR-128 was involved in the alleviation effect of Gs-Rb1 on autophagic flux blockade induced by 3-MCPD. The results showed that Gs-Rb1 inhibited the expression of miR-128 and promoted the nuclear expression and target gene transcription of TFEB. Finally, the findings were confirmed by using a hsa-miR-128 inhibitor and mimic. We found that hsa-miR-128 inhibitor alleviated the autophagic flux blockage and apoptosis caused by 3-MCPD and Gs-Rb1 also had a certain alleviation effect on the autophagic flux blockage and apoptosis caused by hsa-miR-128 mimic. This study elaborated the mechanism by which Gs-Rb1 alleviates hepatotoxicity induced by foodborne 3-MCPD by stimulating autophagic flux via miR-128-targeted TFEB, which provides a reliable theoretical basis and target for the use of natural substances to reduce the harm of food processing pollutants on the human body. PRACTICAL APPLICATION: We found that natural ginsenoside Rb1 can alleviate liver injury induced by 3-MCPD(a toxic substance found in foods such as refined vegetable oil, soy sauce, and baby milk powder), which is conducive to the development and utilization of ginseng and has practical significance for the prevention of foodborne liver injury.
近年来，食源性污染物已成为食品安全领域的热点问题。3-氯-1,2-丙二醇（3-MCPD）是一种广泛存在的食品污染物。在我们之前的研究中，已证实3-MCPD可通过抑制溶酶体功能来阻断自噬流，从而导致肝损伤。人参是一种含有多种生物活性成分的传统中药，其中人参皂苷Rb1（Gs-Rb1）含量最为丰富。在本研究中，我们旨在使用Gs-Rb1改善3-MCPD诱导的自噬流阻断，以减轻肝损伤。首先，通过MTT法筛选确定了Gs-Rb1的无毒剂量，即将Gs-Rb1添加到HepG2细胞中并与3-MCPD共同处理。我们发现Gs-Rb1有效地增强了被3-MCPD抑制的细胞活性。同时，凋亡数据表明Gs-Rb1显著减轻了3-MCPD诱导的HepG2细胞凋亡。随后，通过检测自噬相关蛋白水平和转染mRFP-GFP-LC3腺病毒，我们发现Gs-Rb1可以剂量依赖性方式减轻3-MCPD引起的自噬流阻断。在此基础上，我们使用蛋白质免疫印迹法和定量聚合酶链反应来探究miR-128是否参与了Gs-Rb1对3-MCPD诱导的自噬流阻断的缓解作用。结果表明，Gs-Rb1抑制了miR-128的表达，并促进了转录因子EB（TFEB）的核表达和靶基因转录。最后，通过使用hsa-miR-128抑制剂和模拟物证实了这些发现。我们发现hsa-miR-128抑制剂减轻了3-MCPD引起的自噬流阻断和凋亡，并且Gs-Rb1对hsa-miR-128模拟物引起的自噬流阻断和凋亡也有一定的缓解作用。本研究阐述了Gs-Rb1通过miR-128靶向的TFEB刺激自噬流来减轻食源性3-MCPD诱导的肝毒性的机制，这为利用天然物质减少食品加工污染物对人体的危害提供了可靠的理论依据和靶点。实际应用：我们发现天然人参皂苷Rb1可以减轻3-MCPD（一种存在于精炼植物油、酱油和婴儿奶粉等食品中的有毒物质）诱导的肝损伤，这有利于人参的开发利用，对预防食源性肝损伤具有实际意义。
