miR-194-5p抑制剂对2型糖尿病心肌病小鼠心肌损伤的保护作用及机制探讨

doi:10.11958/20260039

天津医药 ›› 2026, Vol. 54 ›› Issue (6): 577-584.doi: 10.11958/20260039

miR-194-5p抑制剂对2型糖尿病心肌病小鼠心肌损伤的保护作用及机制探讨

张婧¹(), 张明², 卫玮³, 宁海虹³, 韦红梅¹, 李海燕³, 吴宾⁴^,^△()

¹ 新疆军区总医院营养科（邮编830000）
² 甲乳外科
³ 卫勤训练中心
⁴ 核医学科

收稿日期:2026-01-05 修回日期:2026-02-13 出版日期:2026-06-15 发布日期:2026-06-15
通讯作者: ^△E-mail：xjwubin9210@163.com
作者简介:张婧（1982），女，副主任医师，主要从事营养与代谢性心血管疾病的基础与临床方面研究。E-mail：41133178@qq.com
基金资助:
新疆维吾尔自治区自然科学基金面上项目(2023D01C94)

Protective effect and mechanism of miR-194-5p inhibitor on myocardial injury in mice with type 2 diabetic cardiomyopathy

ZHANG Jing¹(), ZHANG Ming², WEI Wei³, NING Haihong³, WEI Hongmei¹, LI Haiyan³, WU Bin⁴^,^△()

¹ Department of Nutrition
² Department of Thyroid and Breast Surgery
³ Medical Service Training Center
⁴ Department of Nuclear Medicine, General Hospital of Xinjiang Military Command, Urumqi 830000, China

Received:2026-01-05 Revised:2026-02-13 Published:2026-06-15 Online:2026-06-15
Contact: ^△E-mail：xjwubin9210@163.com

张婧, 张明, 卫玮, 宁海虹, 韦红梅, 李海燕, 吴宾. miR-194-5p抑制剂对2型糖尿病心肌病小鼠心肌损伤的保护作用及机制探讨[J]. 天津医药, 2026, 54(6): 577-584.

ZHANG Jing, ZHANG Ming, WEI Wei, NING Haihong, WEI Hongmei, LI Haiyan, WU Bin. Protective effect and mechanism of miR-194-5p inhibitor on myocardial injury in mice with type 2 diabetic cardiomyopathy[J]. Tianjin Medical Journal, 2026, 54(6): 577-584.

摘要/Abstract

摘要：

目的探讨miR-194-5p抑制剂（miR-194-5p antagomir）对2型糖尿病心肌病（DCM）小鼠心肌损伤的保护作用及可能机制。方法将60只C57BL/6小鼠随机分为正常对照（NC）组、DCM组、DCM+抑制剂阴性对照（DCM+antagomir control）组、DCM+miR-194-5p抑制剂（DCM+miR-194-5p antagomir）组，每组15只。通过高脂饮食联合链脲佐菌素腹腔注射的方式成功构建2型糖尿病小鼠模型。建模成功后，DCM+miR-194-5p antagomir组小鼠按照10 mg/kg的剂量经尾静脉注射溶解于PBS溶液中的miR-194-5p antagomir，DCM+antagomir control组注射同等剂量的溶解于PBS溶液中的antagomir control，每周2次，持续4周。干预结束后12周行心脏超声检测评估心功能；小麦胚芽凝集素（WGA）染色检测心肌细胞横截面积；酶联免疫吸附试验（ELISA）测定心肌组织炎性因子白细胞介素（IL）-6和肿瘤坏死因子（TNF）-α水平；实时荧光定量聚合酶链反应（RT-qPCR）检测miR-194-5p和双特异性磷酸酶9（DUSP9）mRNA表达；透射电镜观察心肌线粒体超微结构；通过生物信息学方法预测miR-194-5p的靶点并进行富集通路分析；双萤光素酶报告基因实验验证miR-194-5p对DUSP9的靶向调控作用；Western blot法检测DUSP9蛋白及丝裂原活化蛋白激酶（MAPK）通路相关分子表达。结果与NC组相比，DCM组和DCM+antagomir control组左室射血分数（LVEF）和左室短轴缩短率（LVFS）降低，心肌细胞横截面积、凋亡率、炎性因子IL-6和TNF-α水平、磷酸化ERK（p-ERK）、磷酸化p38（p-p38）水平增加（P<0.05），miR-194-5p mRNA表达升高，DUSP9 mRNA和蛋白表达均下降（P<0.05），心肌线粒体超微结构损伤加重。与DCM组和DCM+antagomir control组相比，DCM+miR-194-5p antagomir组LVEF和LVFS增加，心肌细胞横截面积、凋亡率、炎性因子水平、p-ERK、p-p38水平降低（P<0.05），miR-194-5p mRNA表达降低，而DUSP9 mRNA和蛋白表达均升高（P<0.05），心肌线粒体超微结构损伤减轻。生物信息学预测DUSP9是miR-194-5p的潜在靶点，MAPK信号通路居靶基因显著富集的首位。双萤光素酶报告实验证实miR-194-5p可直接靶向DUSP9的3'-UTR。结论抑制miR-194-5p 可改善DCM小鼠心肌损伤，其机制可能与靶向上调DUSP9并进一步抑制MAPK信号通路有关。

关键词: 糖尿病心肌病, 微RNAs, 双特异性磷酸酶类, p38丝裂原活化蛋白激酶类, 细胞凋亡, 细胞外信号调节激酶, miR-194-5p

Abstract:

Objective To investigate the protective effect and its potential mechanism of miR-194-5p inhibitor (miR-194-5p antagomir) on myocardial injury in mice with type 2 diabetic cardiomyopathy (DCM). Methods Sixty C57BL/6 mice were randomly divided into the normal control (NC) group, the DCM group, the DCM+antagomir control group and the DCM+miR-194-5p antagomir group (n=15). The DCM model was established by high-fat diet combined with intraperitoneal injection of streptozotocin. After successful model establishment, mice in the DCM+miR-194-5p antagomir group were injected via the tail vein with miR-194-5p antagomir dissolved in PBS at a dose of 10 mg/kg, while mice in the control group were injected with the same dose of antagomir control dissolved in PBS, twice a week for 4 weeks. Twelve weeks after the intervention, cardiac function was evaluated by echocardiography. Myocardial cell cross-sectional area was detected by wheat germ agglutinin (WGA) staining. Inflammatory cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-α in myocardial tissue were determined by enzyme-linked immunosorbent assay (ELISA). The expression levels of miR-194-5p and dual-specificity phosphatase 9 (DUSP9) mRNA were detected by real-time fluorescence quantitative PCR (RT-qPCR). Myocardial mitochondrial ultrastructure was observed by transmission electron microscopy. The target of miR-194-5p was predicted by bioinformatics methods, and pathway enrichment analysis was performed. The dual-luciferase reporter gene assay was used to verify the targeted regulation of DUSP9 by miR-194-5p. The expression levels of DUSP9 protein and mitogen-activated protein kinase (MAPK) pathway-related molecules were detected by Western blot assay. Results Compared with the NC group, left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were decreased in the DCM group and the DCM+antagomir control group, and myocardial cell cross-sectional area, apoptosis rate, levels of inflammatory factors IL-6 and TNF-α, phosphorylated ERK (p-ERK) and phosphorylated p38 (p-p38) increased (P<0.05). The expression of miR-194-5p mRNA was up-regulated, while the mRNA and protein expression of DUSP9 were down-regulated (P<0.05), and the myocardial mitochondrial ultrastructure was severely damaged. Compared with the DCM group and the DCM+antagomir control group, LVEF and LVFS were increased, and myocardial cell cross-sectional area, apoptosis rate, inflammatory factor levels, p-ERK and p-p38 were decreased in the DCM+miR-194-5p antagomir group (P<0.05). The expression of miR-194-5p mRNA was down-regulated, while the mRNA and protein expression of DUSP9 were up-regulated (P<0.05), and the myocardial mitochondrial ultrastructural injury was alleviated. Bioinformatics prediction indicated that DUSP9 was a potential target of miR-194-5p, and the MAPK signaling pathway was the most significantly enriched pathway among the target genes. The dual-luciferase reporter assay confirmed that miR-194-5p could directly target the 3'-UTR region of DUSP9. Conclusion Inhibition of miR-194-5p attenuates myocardial injury in DCM mice, likely by targeting DUSP9 for upregulation, thereby suppressing the MAPK signaling pathway.

Key words: diabetic cardiomyopathies, microRNAs, dual-specificity phosphatases, p38 mitogen-activated protein kinases, apoptosis, extracellular signal-regulated kinase, miR-194-5p

中图分类号:

R542.2

图/表 13

表1 引物序列

Tab.1 Primers sequence

基因名称	引物序列（5'→3'）	产物大小/bp
miR-194-5p	上游：CGCTGTAACAGCAACTCCAT	75
	下游：GTGCAGGGTCCGAGGT	75
U6	上游：AGCCGAAACAACCAGAGCAC	95
	下游：CAGGTAGGCGATGGTGAAGG	95
DUSP9	上游：GTGTGAGGTGCCTACTTGCTC	201
	下游：GCTCAGTCTTCTGTCCTTGGA	201
β-actin	上游：CATGTACGTTGCTATCCAGGC	181
	下游：CTCCTTAATGTCACGCACGAT	181

表2 各组小鼠的心功能指标比较（n=15，$\stackrel{-}{x}\pm s$）

Tab.2 Comparison of cardiac function indicators between the four groups of mice

组别	LVEF/%		LVFS/%
NC组	73.20±4.34		37.93±2.98
DCM组	56.33±1.97^a		22.67±2.16^a
DCM+antagomir control组	55.79±3.27^a		23.56±3.02^a
DCM+miR-194-5p antagomir组	61.80±2.12^bc		31.29±1.89^bc
F	103.400^＊＊		113.500^＊＊
组别		LVEDD/mm		LVESD/mm
NC组		3.45±0.09		2.51±0.12
DCM组		4.47±0.15^a		3.41±0.09^a
DCM+antagomir control组		4.52±0.24^a		3.49±0.13^a
DCM+miR-194-5p antagomir组		3.98±0.13^bc		3.01±0.11^bc
F		142.700^＊＊		234.700^＊＊

图1 实验小鼠超声心动图成像结果

Fig.1 Echocardiographic imaging results of experimental mice

图2 WGA染色检测各组小鼠心肌横截面积（×400）

Fig.2 Myocardial cross-sectional area detected by WGA staining in each group of mice (×400)

表3 各组小鼠心肌组织炎性因子水平比较（n=15，μg/L，$\stackrel{-}{x}\pm s$）

Tab.3 Comparison of myocardial inflammatory cytokine levels between the four groups of mice

组别	IL-6	TNF-α
NC组	20.47±1.98	9.10±1.21
DCM组	41.38±4.82^a	30.14±3.99^a
DCM+antagomir control组	43.89±3.97^a	29.41±4.02^a
DCM+miR-194-5p antagomir组	28.42±3.41^bc	18.12±2.43^bc
F	134.100^＊＊	153.200^＊＊

图3 不同组别小鼠的心肌组织细胞凋亡情况（TUNEL染色，×400）绿色：TUNEL染色阳性；蓝色：DAPI染色阳性。

Fig.3 Comparison of apoptosis of myocardial tissue between four groups (TUNEL staining, ×400)

表4 各组小鼠心肌组织中miR-194-5p和DUSP9 mRNA表达水平比较（n=6，μg/L，$\stackrel{-}{x}\pm s$）

Tab.4 Comparison of miR-194-5p and DUSP9 mRNA expression between the four groups of mice

组别	miR-194-5p	DUSP9 mRNA
NC组	1.02±0.09	1.00±0.05
DCM组	3.29±0.33^a	0.35±0.03^a
DCM+antagomir control组	3.10±0.21^a	0.41±0.04^a
DCM+miR-194-5p antagomir组	1.42±0.19^bc	0.82±0.07^bc
F	182.200^＊＊	241.500^＊＊

图4 miR-194-5p靶点分析及KEGG通路分析流程图

Fig.4 Flow chart of miR-194-5p target analysis and KEGG analysis

图5 DAVID在线KEGG通路富集分析

Fig.5 DAVID online results of KEGG enrichment pathway analysis

图6 miR-194-5p与DUSP9 3′-UTR的结合位点

Fig. 6 Binding site of miR-194-5p to DUSP9 3′-UTR

图7 透射电镜观察各组小鼠心肌组织中线粒体损伤（×8 000）

Fig.7 Mitochondrial damage in myocardial tissue of mice observed by electron microscopy (×8 000)

图8 Western blot检测不同组别小鼠心肌组织DUSP9、ERK和p38蛋白表达水平 A：NC组；B：DCM组；C：DCM+antagomir control组；D：DCM+miR-194-5p antagomir组。

Fig.8 Western blot analysis of DUSP9, ERK and p38 protein expression levels in myocardial tissue of the four groups of mice

表5 不同组别小鼠心肌组织DUSP9、p-ERK和p-p38蛋白表达比较（n=6，$\stackrel{-}{x}\pm s$）

Tab.5 Comparison of DUSP9, p-ERK and p-p38 protein expression in myocardial tissue between the four groups of mice

组别	DUSP9/GAPDH	p-ERK/ ERK	p-p38/ p38
NC组	1.00±0.08	0.29±0.03	0.31±0.02
DCM组	0.38±0.04^a	0.78±0.10^a	0.81±0.05^a
DCM+antagomir control组	0.42±0.06^a	0.82±0.09^a	0.78±0.09^a
DCM+miR-194-5p antagomir组	0.85±0.07^abc	0.42±0.05^abc	0.41±0.04^abc
F	139.500^＊＊	77.130^＊＊	123.600^＊＊

参考文献 20

[1]	GUAN X, ZHOU C, ZHUO X, et al. The heart of diabetes:unraveling metabolic drivers of cardiomyopathy[J]. Cardiovasc Diabetol Endocrinol Rep, 2025, 11(1):18. doi:10.1186/s40842-025-00231-x.
[2]	KTENOPOULOS N, ANAGNOSTOPOULOU L, APOSTOLOS A, et al. Cellular and molecular pathways in diabetes-associated heart failure:Emerging mechanistic insights and therapeutic opportunities[J]. Curr Issues Mol Biol, 2025, 47(11):886. doi:10.3390/cimb47110886.
[3]	庞志华, 刘丽美, 姚朱华. miR-101-3p的生物学功能及其在心血管疾病中的研究进展[J]. 天津医药, 2025, 53(11):1223-1228.
	PANG Z H, LIU L M, YAO Z H. Research progress of biological functions of miR-101-3p in cardiovascular diseases[J]. Tianjin Med J, 2025, 53(11):1223-1228. doi:10.11958/20252447.
[4]	吴长勇, 孙钺, 保苏丽, 等. 非编码RNAs与心脏纤维化的研究进展[J]. 中国药理学通报, 2023, 39(4):605-609.
	WU C Y, SUN Y, BAO S L, et al. Research progress of non-coding RNAs and cardiac fibrosis[J]. Chinese Pharmacological Bulletin, 2023, 39(4):605-609. doi:10.12360/CPB202205069.
[5]	张婧, 魏玉英, 宁海虹, 等. DUSP9在2型糖尿病心肌病小鼠心肌损伤中的保护作用及其机制[J]. 天津医药, 2026, 54(3):238-244.
	ZHANG J, WEI Y Y, NING H H, et al. Protective effect and mechanism of dual-specificity phosphatase 9 on myocardial injury in mice with type 2 diabetic cardiomyopathy[J]. Tianjin Med J, 2026, 54(3):238-244. doi:10.11958/20253111.
[6]	WANG M, LIANG Y, QIN Y, et al. Active ingredients and mechanism of Gegen Qinlian Decoction in the treatment of diabetic cardiomyopathy:A network pharmacology study[J]. Curr Pharm Des, 2024, 30(36):2896-2910. doi:10.2174/0113816128312242240722080551.
[7]	ZUO G, REN X, QIAN X, et al. Inhibition of JNK and p38 MAPK-mediated inflammation and apoptosis by ivabradine improves cardiac function in streptozotocin-induced diabetic cardiomyopathy[J]. J Cell Physiol, 2019, 234(2):1925-1936. doi:10.1002/jcp.27070.
[8]	张吟, 黄丹丹, 张淑芬, 等. C57BL/6J糖尿病小鼠模型的优化研究[J]. 中国现代应用药学, 2019, 36(6):655-660.
	ZHANG Y, HUANG D D, ZHANG S F, et al. Optimization study on the establishment of diabetic model in C57BL/6J mice[J]. Chinese Modern Applied Pharmacy, 2019, 36(6):655-660. doi:10.13748/j.cnki.issn1007-7693.2019.06.003.
[9]	RABOT S, MEMBREZ M, BRUNEAU A, et al. Germ-free C57BL/6J mice are resistant to high-fat-diet-induced insulin resistance and have altered cholesterol metabolism[J]. FASEB J, 2010, 24(12):4948-4959. doi:10.1096/fj.10-164921.
[10]	ATTRILL E H, SCHARAPOW O, PERERA S, et al. Controlled induction of type 2 diabetes in mice using high fat diet and osmotic-mini pump infused streptozotocin[J]. Sci Rep, 2025, 15(1):8812. doi:10.1038/s41598-025-89162-2.
[11]	BELLEMARE M, BOURCIER L, IGLESIES-GRAU J, et al. Mechanisms of diabetic cardiomyopathy:Focus on inflammation[J]. Diabetes Obes Metab, 2025, 27(5):2326-2338. doi:10.1111/dom.16242.
[12]	张倩, 刘学霖, 张月梅, 等. miRNA在糖尿病心肌病中的靶向调控机制研究进展[J]. 临床心血管病杂志, 2025, 41(2):150-155.
	ZHANG Q, LIU X L, ZHANG Y M, et al. Research progress on the targeted regulatory mechanism of miRNA in diabetic cardiomyopathy[J]. Journal of Clinical Cardiology, 2025, 41(2):150-155. doi:10.13201/j.issn.1001-1439.2025.02.013.
[13]	LIU Y, YUAN J, ZHANG Y, et al. Non-coding RNA as a key regulator and novel target of apoptosis in diabetic cardiomyopathy:Current status and future prospects[J]. Cell Signal, 2025, 128:111632. doi:10.1016/j.cellsig.2025.111632.
[14]	LIU B, WEI Y, HE J, et al. Human umbilical cord-derived mesenchymal stromal cells improve myocardial fibrosis and restore miRNA-133a expression in diabetic cardiomyopathy[J]. Stem Cell Res Ther, 2024, 15(1):120. doi:10.1186/s13287-024-03715-2.
[15]	CAI W, CHONG K, HUANG Y, et al. Empagliflozin improves mitochondrial dysfunction in diabetic cardiomyopathy by modulating ketone body metabolism and oxidative stress[J]. Redox Biol, 2024, 69:103010. doi:10.1016/j.redox.2023.103010.
[16]	PENG C, ZHANG Y, LANG X, et al. Role of mitochondrial metabolic disorder and immune infiltration in diabetic cardiomyopathy: new insights from bioinformatics analysis[J]. J Transl Med, 2023, 21(1):66. doi:10.1186/s12967-023-03928-8.
[17]	MA T, HUANG X, ZHENG H, et al. SFRP2 improves mitochondrial dynamics and mitochondrial biogenesis,oxidative stress,and apoptosis in diabetic cardiomyopathy[J]. Oxid Med Cell Longev, 2021, 2021:9265016. doi:10.1155/2021/9265016.
[18]	WANG J, WEI T, ZHANG W, et al. Inhibition of miR-194-5p avoids DUSP9 downregulation thus limiting sepsis-induced cardiomyopathy[J]. Sci Rep, 2024, 14(1):20313. doi:10.1038/s41598-024-71166-z.
[19]	ZHANG X, JIN Y. DUSP9-mediated inhibition of IRS1/PI3K/AKT pathway contributes to insulin resistance and metabolic dysfunction in gestational diabetes mellitus[J]. Hum Immunol, 2025, 86(3):111263. doi:10.1016/j.humimm.2025.111263.
[20]	LI Z, HUANG Z, LUO Y, et al. DUSP9 alleviates hepatic ischemia/reperfusion injury by restraining both mitogen-activated protein kinase and IKK in an apoptosis signal-regulating kinase 1-dependent manner[J]. Acta Biochim Biophys Sin(Shanghai), 2022, 54(12):1811-1821. doi:10.3724/abbs.2022183.

miR-194-5p抑制剂对2型糖尿病心肌病小鼠心肌损伤的保护作用及机制探讨

Protective effect and mechanism of miR-194-5p inhibitor on myocardial injury in mice with type 2 diabetic cardiomyopathy

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