MiR345-5p在金属镉诱导急性肝脏损伤中的作用研究

摘要/Abstract

摘要：

目的了解微小RNA在金属镉致急性肝脏损伤中的调控机制，筛查镉致肝脏损伤的微小RNA的关键分子。方法将C57BL/6雄性小鼠随机分为2组，一组为对照组，另一组为镉染毒组，对染毒组使用2 mg/kg 氯化镉溶液进行腹腔注射，对照组使用生理盐水，染毒结束后，收集血清及肝脏组织，评估其肝脏损伤程度，并检测miR345-5p及其靶基因GSTM2表达水平。同时用金属镉处理L02细胞，观察染毒后细胞活力变化及miR345-5p及其靶基因GSTM2表达水平，利用双荧光素酶报告系统验证miR345-5p对靶基因GSTM2的调控机制。结果金属镉染毒后，小鼠肝脏出现急性炎症反应，肝脏中miR345-5p表达增加6.85倍，靶基因GSTM2表达降低0.64倍。L02细胞水平，miR345-5p及靶基因GSTM2与金属镉染毒呈现剂量反应关系。并且加入miR345-5p mimic后，GSTM2 mRNA表达下降，镉染毒组的细胞活力下降72.38%。双荧光素酶报告系统显示加入miR345-5p mimic后，野生组荧光强度降低，突变组荧光强度不变。结论 miR345-5p可通过介导GSTM2表达下调加重金属镉诱导急性肝脏损伤。

关键词: 微小RNA, 急性肝脏损伤, 镉, miR-345-5p, 谷胱甘肽S-转移酶, 二相解毒酶

Abstract:

Objective To investigate the regulatory mechanism of miRNA in metallic cadmium-induced acute liver injury，screen key molecules of miRNAs associated with cadmium induced liver injury. Methods Male C57BL/6 mice were randomly divided into a control group and a cadmium-exposed group. The exposed group received intraperitoneal injections of 2 mg/kg cadmium chloride solution，while the control group was administered normal saline. The serum and liver tissues were collected post-exposure to evaluate liver injury severity and measure the expression levels of miR345-5p and its target gene GSTM2. L02 cells were treated with metallic cadmium to assess changes in cell viability and the expression of miR345-5p and GSTM2. The dual-luciferase reporter system was employed to validate miR345-5p's regulatory effect on GSTM2. Results The metallic cadmium exposure induced acute inflammatory responses in mouse livers，with miR345-5p expression increasing by 6.85-fold and GSTM2 expression decreasing by 0.64-fold. In L02 cells，miR345-5p and GSTM2 exhibited a dose-response relationship to cadmium exposure. Transfection with miR345-5p mimic reduced the mRNA expression of GSTM2 and decreased cell viability by 72.38% in cadmium-treated cells. The dual-luciferase assay confirmed that miR345-5p mimic significantly reduced fluorescence intensity in the wild-type group but not in the mutant group. Conclusion The miR345-5p exacerbates metallic cadmium-induced acute liver injury by downregulating GSTM2 expression.

Key words: MicroRNA, Acute Liver Injury, Cadmium, miR-345-5p, Glutathione S-transferase, Phase Ⅱdetoxification enzyme

中图分类号:

R-332

柳晓玲, 羊晶晶, 林凌鸿. MiR345-5p在金属镉诱导急性肝脏损伤中的作用研究. [J]职业与健康, 2026, 42(11): 1475-1479.

LIU Xiaoling, YANG Jingjing, LIN Linghong. Study on role of miR345-5p in metallic cadmium-induced acute liver injury. [J]OCCUPATION AND HEALTH, 2026, 42(11): 1475-1479.

图/表 5

图1 急性氯化镉染毒内暴露指标检测注：a与对照组相比，P<0.05。

图2 急性氯化镉染毒后小鼠血液生化指标注：ALT—丙氨酸转氨酶；AST—天冬氨酸转氨酶；a与对照组相比，P<0.05。

图3 急性氯化镉染毒小鼠肝脏（HE染色）

图4 小鼠肝脏中miR-345-5p及其靶基因表达注：GSTM2—谷胱甘肽S-转移酶；a与对照组相比，P<0.05。

图5 镉染毒L02细胞体外实验结注：图A—MTT法检测L02细胞活力；图B~C—利用RT-QPCR技术检测各组miR-345-5p与GSTM2表达；图D—加入miR-345-5p mimic模拟物后，对照组与镉染毒组细胞活力情况；图E—对照组与miR-345-5p mimic组GSTM2基因mRNA表达；图F—采用双荧光素酶报告系统对野生组、突变组的荧光强度；a与对照组相比，P<0.05。

参考文献 16

[1]	NORDBERG G F. Historical perspectives on cadmium toxicology[J]. TAAP, 2009, 238(3):192-200.
[2]	张晨阳, 严俊, 王梅, 等. 低水平镉暴露对人体免疫学指标的影响[J]. 中国预防医学杂志, 2022, 23(7):481-485.
[3]	易宗娓, 何作顺. 镉污染与痛痛病[J]. 职业与健康, 2014, 30(17):2511-2513.
[4]	王序兵, 赵艳洁, 于典科. 微小RNA在酒精致肝损伤中的作用机制研究进展[J]. 环境与职业医学, 2020, 37(4):406-411.
[5]	骈雅婧, 罗皓珑, 林芮, 等. miRNA转录组学分析镉致小鼠睾丸间质细胞毒性作用机制[J]. 卫生研究, 2023, 52(3):483-488,505.
[6]	JIANG X, LI W, TAN M, et al. Identification of miRNAs involved in liver injury induced by chronic exposure to cadmium[J]. Toxicology, 2022, 469(1):153,133.
[7]	SENGUL E, YILDIRIM S, CINAR İ, et al. Mitigation of acute hepatotoxicity induced by cadmium through morin:Modulation of oxidative and pro-apoptotic endoplasmic reticulum stress and inflammatory responses in rats[J]. Biol Trace Elem Res, 2024, 202(11):5106-5117. DOI
[8]	李雪, 王健, 刘敏, 等. 线粒体自噬在镉致小鼠肝炎症中的作用及NF-κB通路调控[J]. 环境与健康杂志, 2024, 41(5):385-390.
[9]	刘敏, 陈春梅, 谭聪, 等. 环境表观遗传学研究进展[J]. 环境卫生学杂志, 2011, 1(5):35-41.
[10]	QU Z, DENG L, FENG C, et al. Environmental cadmium-induced circRNA-miRNA-mRNA network regulatory mechanism in human normal liver cell model[J]. CRT, 2024, 24(7):1-10.
[11]	杨忠海, 冯文涛, 张大鹏, 等. miR-345-5p,chemerin,HDL-C在HLAP患者中的表达及与HLAP发病的相关性研究[J]. 国际检验医学杂志, 2023, 44(6):641-644.
[12]	李昌浩. 饮水镉暴露加重非酒精性脂肪肝模型小鼠肝损伤及其机制研究[D]. 苏州: 苏州大学, 2023.
[13]	CHEN W, ZhAO W, YANG A, et al. Integrated analysis of microRNA and gene expression profiles reveals a functional regulatory module associated with liver fibrosis[J]. Gene, 2017, 636(1):87-95. DOI URL
[14]	LAN T, HU Y, HU F, et al. Hepatocyte glutathione S-transferase mu 2 prevents non-alcoholic steatohepatitis by suppressing ASK1 signaling[J]. J Hepatol, 2022, 76(2):407-419. DOI URL
[15]	MUHAMMAD I, WANG H, SUN X, et al. Dual role of dietary curcumin through attenuating afb1-induced oxidative stress and liver injury via modulating liver phase-I and phase-II enzymes involved in AFB1 bioactivation and detoxification[J]. Front Pharmacol, 2018, 9(1):554-564. DOI URL
[16]	姚芹, 吴顺华, 汤红英, 等. 亚砷酸钠亚慢性染毒对小鼠体内谷胱甘肽S-转移酶活力及其基因表达的影响[J]. 新疆医科大学学报, 2010, 33(12):1385-1387.