槲皮素通过调控L型钙通道改善糖尿病大鼠心肌损伤

doi:10.12122/j.issn.1673-4254.2025.03.11

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (3): 531-541.doi: 10.12122/j.issn.1673-4254.2025.03.11

• • 上一篇

槲皮素通过调控L型钙通道改善糖尿病大鼠心肌损伤

孙红燕¹^,²(), 卢国庆¹^,², 付程文¹^,², 徐梦文³, 朱小翌³, 邢国权³, 刘乐强³, 柯雨菲³, 崔乐妹³, 陈睿旸³, 王磊¹^,², 康品方¹^,²(), 唐碧¹^,²

^1.蚌埠医科大学，第一附属医院心血管内科，安徽蚌埠 233000
^2.蚌埠医科大学，心脑血管病研究中心生理学教研室，安徽蚌埠 233000
^3.蚌埠医科大学，临床医学院，安徽蚌埠 233000

收稿日期:2024-09-29 出版日期:2025-03-20 发布日期:2025-03-28
通讯作者: 康品方,唐碧 E-mail:1210499483@qq.com;kangpinfang.1016@163.com
作者简介:孙红燕，在读硕士研究生，E-mail: 1210499483@qq.com
基金资助:
国家自然科学基金(81970313);安徽省自然科学基金(2208085MH192);安徽省临床医学研究转化项目(202304295107020087);安徽省高校协同创新项目(GXXT-2020-019);安徽省高校优秀青年项目(2022AH030141);蚌埠医学院研究生科研创新计划(Byycxz23040)

Quercetin ameliorates myocardial injury in diabetic rats by regulating L-type calcium channels

Hongyan SUN¹^,²(), Guoqing LU¹^,², Chengwen FU¹^,², Mengwen XU³, Xiaoyi ZHU³, Guoquan XING³, Leqiang LIU³, Yufei KE³, Lemei CUI³, Ruiyang CHEN³, Lei WANG¹^,², Pinfang KANG¹^,²(), Bi TANG¹^,²

^1.Department of Cardiovascular Medicine, First Affiliated Hospital, Bengbu Medical University, Bengbu 233000, China
^2.Department of Physiology, Cardiovascular and Cerebrovascular Disease Research Center, Bengbu Medical University, Bengbu 233000, China
^3.College of Clinical Medicine, Bengbu Medical University, Bengbu 233000, China

Received:2024-09-29 Online:2025-03-20 Published:2025-03-28
Contact: Pinfang KANG, Bi TANG E-mail:1210499483@qq.com;kangpinfang.1016@163.com
Supported by:
National Natural Science Foundation of China(81970313)

摘要/Abstract

摘要：

目的探讨槲皮素对糖尿病大鼠心肌铜死亡与心肌L型钙电流的影响。方法动物实验：对40只SD大鼠进行对照（Con）和糖尿病模型构建，后者通过高糖高脂饲料且用STZ诱导，糖尿病模型进一步分为模型组（DM）、槲皮素组（DM+Que）和恩格列净组（DM+Empa），n=10。每2周测量大鼠血糖和体质量，通过超声心动图评估心脏功能，通过HE染色、天狼猩红染色和WGA染色观察心肌组织形态学变化，并检测血清铜离子水平和FDX1的表达情况。体外细胞（大鼠H9c2心肌细胞）实验：分别设置低糖组（Con）、高糖组（HG）、槲皮素组（HG+Que）、伊利司莫组（HG+ES）和伊利司莫+槲皮素组（HG+ES+Que），通过CK-MB和LDH评估心肌损伤，检测FDX1蛋白表达。膜片钳技术检测心肌细胞L钙电流变化。结果与Con组相比，DM组大鼠血糖增加（P<0.05），体质量下降（P<0.05），左心功能下降，血清铜水平和FDX1表达增加（P<0.01），心肌L钙电流减小（P<0.01），动作电位时程延长（P<0.01）。与DM组相比，药物组血糖水平降低（P<0.05）、体质量增加（P<0.05）、左心功能改善。与DM+Empa组相比，槲皮素组铜离子水平、FDX1表达减少（P<0.01），心肌L钙电流增加（P<0.05）。在体外实验中，HG组的FDX1表达水平和心肌损伤均增加，槲皮素干预后，FDX1表达降低和心肌损伤改善。HG+ES组的FDX1、CK-MB和LDH均升高（P<0.05），加入槲皮素后，这些指标与HG+ES组相比差异没有统计学意义（P>0.05）。结论槲皮素可以改善糖尿病大鼠心肌损伤，可能与抑制铜死亡信号通路从而恢复心肌L型钙电流有关。

关键词: 槲皮素, 糖尿病心肌病, L-型钙电流, 铜死亡

Abstract:

Objective To investigate the effects of quercetin on cuproptosis and L-type calcium currents in the myocardium of diabetic rats. Methods Forty SD rats were randomized into control group and diabetic model groups. The rat models of diabetes mellitus (DM) induced by high-fat and high-sugar diet combined with streptozotocin (STZ) injection were further divided into DM model group, quercetin treatment group, and empagliflozin treatment group (n=10). Blood glucose and body weight were measured every other week, and cardiac function of the rats was evaluated using echocardiography. HE staining, Sirius red staining, and wheat germ agglutinin (WGA) analysis were used to observe the changes in myocardial histomorphology, and serum copper levels and myocardial FDX1 expression were detected. In cultured rat cardiomyocyte H9c2 cells with high-glucose exposure, the effects of quercetin and elesclomol, alone or in combination, on intracellular CK-MB and LDH levels and FDX1 expression were assessed, and the changes in L-type calcium currents were analyzed using patch-clamp technique. Results The diabetic rats exhibited elevated blood glucose, reduced body weight, impaired left ventricular function, increased serum copper levels and myocardial FDX1 expression, decreased L-type calcium currents, and prolonged action potential duration. Quercetin and empagliflozin treatment significantly lowered blood glucose, improved body weight, and restored cardiac function of the diabetic rats, and compared with empagliflozin, quercetin more effectively reduced serum copper levels, downregulated FDX1 expression, and enhanced myocardial L-type calcium currents in diabetic rats. In H9c2 cells, high glucose exposure significantly increased myocardial expressions of FDX1, CK-MB and LDH, which were effectively lowered by quercetin treatment; Elesclomol further elevated FDX1, CK-MB and LDH levels in the exposed cells, and these changes were not significantly affected by the application of quercetin. Conclusion Quercetin ameliorates myocardial injury in diabetic rats possibly by suppressing myocardial cuproptosis signaling and restoring L-type calcium channel activity.

Key words: quercetin, diabetic cardiomyopathy, L-type calcium currents, cuproptosis

孙红燕, 卢国庆, 付程文, 徐梦文, 朱小翌, 邢国权, 刘乐强, 柯雨菲, 崔乐妹, 陈睿旸, 王磊, 康品方, 唐碧. 槲皮素通过调控L型钙通道改善糖尿病大鼠心肌损伤[J]. 南方医科大学学报, 2025, 45(3): 531-541.

Hongyan SUN, Guoqing LU, Chengwen FU, Mengwen XU, Xiaoyi ZHU, Guoquan XING, Leqiang LIU, Yufei KE, Lemei CUI, Ruiyang CHEN, Lei WANG, Pinfang KANG, Bi TANG. Quercetin ameliorates myocardial injury in diabetic rats by regulating L-type calcium channels[J]. Journal of Southern Medical University, 2025, 45(3): 531-541.

图/表 6

图1 槲皮素对糖尿病大鼠血糖、体质量的影响

Fig.1 Effect of quercetin on blood glucose (A) and weight (B) in diabetic rats (Mean±SD, n=6). **P<0.01 vs Con, ##P<0.01 vs DM.

图2 槲皮素对糖尿病大鼠心功能的影响

Fig.2 Effect of quercetin on cardiac function in diabetic rats. A: Comparison of echocardiographic findings in each group. B-G: LVEF%, LVFS%, LVEDD, LVESD, CO and HR of SD rats in each group. Data are presented as Mean±SD (n=6), **P<0.01 vs Con; #P<0.05, ##P<0.01 vs DM.

图3 槲皮素对糖尿病大鼠心肌结构的影响

Fig.3 Effect of quercetin on myocardial structure in diabetic rats. A: HE, Sirius Red, and WGA staining results of SD rats in each group. B: Analysis of the results of Sirius Red staining in each group. C: Analysis of WGA staining in each group. Data are presented as Mean±SD (n=3). **P<0.01 vs Con, #P<0.05, ##P<0.01 vs DM.

图4 槲皮素对糖尿病大鼠心肌中FDX1及血清铜的影响

Fig.4 Effect of quercetin on myocardial FDX1 expression and serum copper level in diabetic rats. A: Immunohistochemical staining of FDX1. B: Quantitative analysis of FDX1 expression levels; C: Serum Cu levels in each group. D, E: FDX1 protein expression levels of SD rats in different groups detected by Western blotting. Data are presented as Mean±SD (n=3). **P<0.01 vs Con; #P<0.05, ##P<0.01 vs DM; △P<0.05, △△P<0.01 vs DM+Empa.

图5 槲皮素对高糖诱导的H9C2心肌细胞损伤的影响

Fig.5 Effect of quercetin on high glucose-induced injury in rat H9C2 cardiomyocytes. A, B: FDX1 protein expression levels of FDX1 protein in each group detected by Westernblotting. C, D: FDX1 expression levels in each group detected by immunofluorescence assay. E, F: CK-MB and LDH levels in each group. Data are presented as Mean±SD (n=3). **P<0.01 vs Con; #P<0.05, ##P<0.01 vs HG.

图6 槲皮素对糖尿病大鼠心肌ICa-L和动作电位(AP)的影响

Fig.6 Effects of quercetin on myocardial ICa-L and action potentials (AP) in diabetic rats. A: Representative recording traces ICa-L currents in different groups. B: Current-voltage relationship of ICa-L currents in different groups. C: Representative action potentials (AP) in APD60 and APD90 repolarization and APA recorded in different groups. D, E: APDs and APA in different groups. Data are presented as Mean±SD (n=12), *P<0.05, **P<0.01 vs Con, #P<0.05, ##P<0.01 vs DM, △P<0.05 vs DM+Empa.

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槲皮素通过调控L型钙通道改善糖尿病大鼠心肌损伤

Quercetin ameliorates myocardial injury in diabetic rats by regulating L-type calcium channels

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