miR-493-3p通过lncRNA CCAT2/TCF7L2轴对高糖诱导的心肌细胞损伤和凋亡的影响

doi:10.11958/20252647

天津医药 ›› 2026, Vol. 54 ›› Issue (4): 344-351.doi: 10.11958/20252647

miR-493-3p通过lncRNA CCAT2/TCF7L2轴对高糖诱导的心肌细胞损伤和凋亡的影响

宋金萍(), 崔彦杰, 王昌敏^△()

新疆维吾尔自治区人民医院临床检验中心（邮编830001）

收稿日期:2025-08-08 修回日期:2025-12-15 出版日期:2026-04-15 发布日期:2026-04-14
通讯作者: ^△E-mail：wcm224@126.com
作者简介:宋金萍（1983），女，副研究员，主要从事分子诊断方面研究。E-mail：songjinping2022@163.com
基金资助:
新疆维吾尔自治区自然科学基金项目(2022D01C617)

The effect of miR-493-3p on high glucose-induced myocardial cell injury and apoptosis via the lncRNA CCAT2/TCF7L2 axis

SONG Jinping(), CUI Yanjie, WANG Changmin^△()

Clinical Laboratory Center, People's Hospital of Xinjiang Uygur Autonomous Region, Urumqi 830001, China

Received:2025-08-08 Revised:2025-12-15 Published:2026-04-15 Online:2026-04-14
Contact: ^△E-mail：wcm224@126.com

宋金萍, 崔彦杰, 王昌敏. miR-493-3p通过lncRNA CCAT2/TCF7L2轴对高糖诱导的心肌细胞损伤和凋亡的影响[J]. 天津医药, 2026, 54(4): 344-351.

SONG Jinping, CUI Yanjie, WANG Changmin. The effect of miR-493-3p on high glucose-induced myocardial cell injury and apoptosis via the lncRNA CCAT2/TCF7L2 axis[J]. Tianjin Medical Journal, 2026, 54(4): 344-351.

摘要/Abstract

摘要：

目的探讨miR-493-3p通过调节长链非编码RNA（lncRNA）结肠癌相关转录物2（CCAT2）/转录因子7样2（TCF7L2）轴对高糖（HG）诱导的心肌细胞损伤和凋亡的影响。方法用高糖（55 mmol/L葡萄糖）培养人心肌细胞AC16，体外构建糖尿病心肌病（DCM）细胞模型。实验分组：Control组、HG组、HG+mimics-NC组、HG+miR-493-3p-mimics组、HG+干扰RNA阴性对照（Smart Silencer-NC）组、HG+干扰RNA CCAT2（Smart Silencer-CCAT2）组、HG+miR-493-3p-mimics+LV-NC组、HG+miR-493-3p-mimics+LV-CCAT2组。CCK-8法及流式细胞术检测细胞活力和凋亡率；试剂盒法检测细胞培养液中乳酸脱氢酶（LDH）和天冬氨酸转氨酶（AST）及细胞中超氧化物歧化酶（SOD）水平；双萤光素酶报告基因实验验证miR-493-3p和CCAT2之间的靶向关系；实时荧光定量PCR（qRT-PCR）检测miR-493-3p、lncRNA CCAT2、β-catenin、TCF7L2、胱天蛋白酶（Caspase）-3、细胞周期蛋白（Cyclin）D1 mRNA表达水平；Western blot检测β-catenin（核）、β-catenin（胞浆）、TCF7L2、Caspase-3、CyclinD1蛋白表达。结果与Control组相比，HG组miR-493-3p、OD₄₅₀值、SOD水平降低，lncRNA CCAT2、LDH、AST水平、细胞凋亡率、TCF7L2 mRNA和蛋白表达水平升高，上调miR-493-3p或敲低lncRNA CCAT2表达可拮抗上述变化（P<0.05）。双萤光素酶报告基因实验验证了miR-493-3p与lncRNA CCAT2和TCF7L2的靶向关系。与HG+miR-493-3p-mimics组比较，HG+miR-493-3p-mimics+LV-CCAT2组TCF7L2、β-catenin（核）及下游Caspase-3和CyclinD1 mRNA和蛋白表达均升高（P<0.05）。结论 miR-493-3p可通过抑制lncRNA CCAT2的表达减缓高糖条件下心肌细胞损伤和凋亡，二者对TCF7L2的调控存在竞争关系。

关键词: 糖尿病心肌病, 肌细胞, 心脏, 转录因子7样2蛋白, 细胞凋亡, miR-493-3p, LncRNA CCAT2

Abstract:

Objective To investigate the impact of miR-493-3p on high glucose-induced cardiomyocyte injury and apoptosis by regulating long noncoding RNA (lncCCAT2)/transcription factor 7-like 2 (TCF7L2) axis. Methods Diabetic cardiomyopathy (DCM) cell model (AC16) was established in vitro with high glucose (HG, 55 mmol/L glucose). Experimental grouping:the control group, the HG group, the si-RNA negative control group, the si-CCAT2 group, the RNA mimics negative control group, the miR-493-3p-mimics group, the miR-493-3p-mimics +Lentivirus negative control group and the miR-493-3p-mimics +LV-CCAT2 group. Cell viability and apoptosis rate were detected by CCK8 assay and flow cytometry. The levels of lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) in cell culture medium and superoxide dismutase (SOD) in cells were detected by the kit methods. Dual luciferase reporter gene assay was used to verify the targeting relationship between miR-493-3p and lncRNA CCAT2. The expression of miR-493-3p, lncRNA CCAT2, β?catenin, TCF7L2, Caspase-3 and CyclinD1 genes were detected by RT-PCR. Western blot assay was used to detect the expressions of β-catenin (nuclear), β-catenin (Cytoplasm), TCF7L2, Caspase-3 and CyclinD1 proteins. Results Compared with the control group, miR-493-3p, OD₄₅₀and SOD were decreased in the HG group, and levels of lncRNA CCAT2, LDH, AST, apoptosis rate, the expression levels of TCF7L2 mRNA and protein increased. Up-regulating miR-493-3p or knocking down the expression of lncRNA CCAT2 can antagonize the above changes (P<0.05). The double luciferase reporter gene experiment verified the relationship between miR-493-3p and CCAT2 and TCF7L2. Compared with the HG+miR-493-3p-mimics group, the TCF7L2, β-catenin (nuclear), downstream Caspase-3 and CyclinD1 mRNA and protein expression were increased in the HG+miR-493-3p-mimics+LV-CCAT2 group (P<0.05). Conclusion miR-493-3p can alleviate myocardial cell injury and apoptosis under high glucose condition by inhibiting lncRNA CCAT2 expression, and there is a competitive relationship between the two regulation of TCF7L2.

Key words: diabetic cardiomyopathies, myocytes, cardiac, transcription factor 7-like 2 protein, apoptosis, miR-493-3p, LncRNA CCAT2

中图分类号:

R542.2，R587.1

图/表 13

参考文献 18

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基因名称	引物序列（5'→3'）	产物大小/bp
miR-493-3p	上游：GTGAAGGTCTACTGTGTGC	97
miR-493-3p	下游：GTCCAGTTTTTTTTTTTTCCTG	97
CCAT2	上游：CTTCCAGCTCCACCTCTG	158
CCAT2	下游：GAGCTCAAAGGACGATGAGG	158
β-catenin	上游：AAAGCGGCTGTTAGTCACTGG	215
β-catenin	下游：CGAGTCATTGCATACTGTCCAT	215
TCF7L2	上游：GAAGGAGCGACAGCTTCATA	200
TCF7L2	下游：GGGGGAGGCGAATCTAGTAA	200
Caspase-3	上游：GAAATTGTGGAATTGATGCGTGA	164
Caspase-3	下游：CTACAACGATCCCCTCTGAAAAA	164
CyclinD1	上游：GGAGGAGGGTTGTGCTACAG	289
CyclinD1	下游：GCCTAGAACCCCACTACAGC	289
U6 snRNA	上游：CTCAACTGGTGTCGTGGAGTCGGC AATTC	102
U6 snRNA	下游：ACACTCCAGCTGGGTCACCGGGTGT AAATC	102
GAPDH	上游：TGTTGCCATCAATGACCCCTT	202
GAPDH	下游：CTCCACGACGTACTCAGCG	202

基因名称	引物序列（5'→3'）	产物大小/bp
miR-493-3p	上游：GTGAAGGTCTACTGTGTGC	97
miR-493-3p	下游：GTCCAGTTTTTTTTTTTTCCTG	97
CCAT2	上游：CTTCCAGCTCCACCTCTG	158
CCAT2	下游：GAGCTCAAAGGACGATGAGG	158
β-catenin	上游：AAAGCGGCTGTTAGTCACTGG	215
β-catenin	下游：CGAGTCATTGCATACTGTCCAT	215
TCF7L2	上游：GAAGGAGCGACAGCTTCATA	200
TCF7L2	下游：GGGGGAGGCGAATCTAGTAA	200
Caspase-3	上游：GAAATTGTGGAATTGATGCGTGA	164
Caspase-3	下游：CTACAACGATCCCCTCTGAAAAA	164
CyclinD1	上游：GGAGGAGGGTTGTGCTACAG	289
CyclinD1	下游：GCCTAGAACCCCACTACAGC	289
U6 snRNA	上游：CTCAACTGGTGTCGTGGAGTCGGC AATTC	102
U6 snRNA	下游：ACACTCCAGCTGGGTCACCGGGTGT AAATC	102
GAPDH	上游：TGTTGCCATCAATGACCCCTT	202
GAPDH	下游：CTCCACGACGTACTCAGCG	202

组别	miR-493-3p	lncRNA CCAT2
Control组	1.01±0.15	1.01±0.12
HG组	0.30±0.02^a	2.24±0.38^a
HG+mimics-NC组	0.31±0.03	2.07±0.21
HG+miR-493-3p-mimics组	0.92±0.08^bc	1.37±0.05^bc
HG+Smart Silencer-NC组	0.36±0.07	2.08±0.23
HG+Smart Silencer-CCAT2组	1.14±0.16^bd	0.94±0.21^bd
F	130.960^＊＊	58.594^＊＊

组别	miR-493-3p	lncRNA CCAT2
Control组	1.01±0.15	1.01±0.12
HG组	0.30±0.02^a	2.24±0.38^a
HG+mimics-NC组	0.31±0.03	2.07±0.21
HG+miR-493-3p-mimics组	0.92±0.08^bc	1.37±0.05^bc
HG+Smart Silencer-NC组	0.36±0.07	2.08±0.23
HG+Smart Silencer-CCAT2组	1.14±0.16^bd	0.94±0.21^bd
F	130.960^＊＊	58.594^＊＊

组别	OD₄₅₀值			LDH/（U/L）
Control组	0.89±0.03			89.47±17.99
HG组	0.63±0.03^a			396.68±43.54^a
HG+mimics-NC组	0.66±0.04			374.51±78.11
HG+miR-493-3p-mimics组	0.76±0.04^bc			176.56±35.66^bc
HG+Smart Silencer-NC组	0.62±0.04			330.88±36.32
HG+Smart Silencer-CCAT2组	0.75±0.05^bd			234.03±18.54^bd
F	585.245^＊＊			20.861^＊＊
组别		AST/ （U/L）	SOD/ （U/mgprot）		凋亡率/ %
Control组		13.90±3.48	16.58±1.98		2.09±0.46
HG组		76.49±18.98^a	3.73±0.82^a		13.62±1.12^a
HG+mimics-NC组		71.89±11.50	3.98±0.82		13.26±1.12
HG+miR-493-3p-mimics组		26.79±6.50^bc	9.63±0.56^bc		7.24±0.56^bc
HG+Smart Silencer-NC组		69.03±12.08	4.09±0.96		12.77±1.10
HG+Smart Silencer-CCAT2组		36.69±4.91^bd	11.39±1.93^bd		8.04±0.51^bd
F		7.050^＊	52.465^＊＊		78.865^＊＊

miR-493-3p通过lncRNA CCAT2/TCF7L2轴对高糖诱导的心肌细胞损伤和凋亡的影响

The effect of miR-493-3p on high glucose-induced myocardial cell injury and apoptosis via the lncRNA CCAT2/TCF7L2 axis

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参考文献 18

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组别	mRNA	蛋白
Control组	1.01±0.08	1.00±0.05
HG组	1.85±0.04^a	1.64±0.10^a
HG+mimics-NC组	1.80±0.07	1.53±0.11
HG+miR-493-3p-mimics组	1.21±0.09^bc	1.18±0.09^bc
HG+Smart Silencer-NC组	2.18±0.36	1.81±0.19
HG+Smart Silencer-CCAT2组	1.47±0.18^bd	1.29±0.16^bd
F	3.923^＊	4.011^＊

组别	miR-493-3p	CCAT2
Control组	1.01±0.15	1.01±0.13
HG组	0.33±0.04^a	2.04±0.30^a
HG+mimics-NC组	0.30±0.06	2.10±0.12
HG+miR-493-3p-mimics组	0.92±0.08^bc	1.37±0.05^bc
HG+miR-493-3p-mimics+ LV-NC组	0.89±0.07	1.44±0.18
HG+miR-493-3p-mimics+ LV-CCAT2组	0.44±0.07^d	1.92±0.27^d
F	48.739^＊＊	13.234^＊＊
组别	OD₄₅₀值	凋亡率/%
Control组	0.88±0.04	2.14±0.52
HG组	0.64±0.03^a	13.28±1.07^a
HG+mimics-NC组	0.65±0.03	13.44±1.21
HG+miR-493-3p-mimics组	0.76±0.04^bc	7.24±0.56^bc
HG+miR-493-3p-mimics+ LV-NC组	0.75±0.03	7.42±0.51
HG+miR-493-3p-mimics+ LV-CCAT2组	0.66±0.04^d	10.02±0.91^bd
F	36.704^＊＊	78.865^＊＊

组别	LDH/ （U/L）	AST/ （U/L）	SOD/ （U/mgprot）
Control组	83.15±18.17	15.74±1.95	14.21±2.47
HG组	415.67±40.31^a	74.00±16.87^a	3.73±0.74^a
HG+mimics-NC组	403.80±63.98	76.49±11.71	3.90±0.32
HG+miR-493-3p-mimics组	176.56±35.66^bc	26.79±6.50^bc	9.63±0.56^bc
HG+miR-493-3p-mimics+ LV-NC组	168.65±30.12	29.55±8.94	9.27±1.36
HG+miR-493-3p-mimics+ LV-CCAT2组	342.83±55.47^d	60.38±12.15^d	4.27±0.31^d
F	20.681^＊＊	7.050^＊＊	52.465^＊＊

组别	TCF7L2	β-catenin
Control组	0.99±0.09	1.09±0.06
HG组	1.87±0.01^a	1.85±0.05^a
HG+mimics-NC组	1.82±0.04	1.77±0.02
HG+miR-493-3p-mimics组	1.21±0.09^bc	1.08±0.09^bc
HG+miR-493-3p-mimics+ LV-NC组	1.25±0.01	1.10±0.05
HG+miR-493-3p-mimics+ LV-CCAT2组	1.73±0.11^d	1.68±0.08^d
F	56.535^＊＊	54.914^＊＊
组别	Caspase-3	CyclinD1
Control组	1.05±0.12	1.02±0.08
HG组	2.20±0.08^a	1.81±0.14^a
HG+mimics-NC组	2.27±0.09	1.86±0.18
HG+miR-493-3p-mimics组	1.25±0.09^bc	1.22±0.22^bc
HG+miR-493-3p-mimics+ LV-NC组	1.24±0.13	1.25±0.10
HG+miR-493-3p-mimics+ LV-CCAT2组	1.93±0.11^d	1.64±0.10^d
F	64.306^＊＊	16.067^＊＊

组别		TCF7L2		β-catenin（核）
Control组		0.39±0.02		0.38±0.03
HG组		0.61±0.04^a		0.87±0.06^a
HG+mimics-NC组		0.60±0.05		0.88±0.06
HG+miR-493-3p-mimics组		0.44±0.03^bc		0.55±0.11^bc
HG+miR-493-3p-mimics+ LV-NC组		0.47±0.02		0.55±0.06
HG+miR-493-3p-mimics+ LV-CCAT2组		0.56±0.03^d		0.82±0.05^d
F		24.892^＊＊		55.854^＊＊
组别	β-catenin（胞浆）		Caspase-3		CyclinD1
Control组	0.52±0.03		0.24±0.03		0.29±0.01
HG组	0.35±0.03^a		0.66±0.03^a		0.63±0.08^a
HG+mimics-NC组	0.34±0.02		0.63±0.02		0.61±0.09
HG+miR-493-3p-mimics组	0.47±0.04^bc		0.44±0.01^bc		0.43±0.02^bc
HG+miR-493-3p-mimics+ LV-NC组	0.47±0.02		0.47±0.03		0.42±0.06
HG+miR-493-3p-mimics+ LV-CCAT2组	0.32±0.02^d		0.61±0.02^d		0.58±0.07^d
F	37.876^＊＊		131.691^＊＊		11.536^＊＊

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