[1]宋彦庆,杜章辉,丁冰钰.水苏碱调节Hippo-YAP信号通路对高糖诱导心肌细胞损伤模型影响的实验研究[J].现代检验医学杂志,2025,40(04):73-78.[doi:10.3969/j.issn.1671-7414.2025.04.013]
　SONG Yanqing,DU Zhanghui,DING Bingyu.Experimental Study on the Effect of Stachydrine on Myocardial Cell Injury Model Induced by High Glucose Through Regulating Hippo-YAP Signaling Pathway[J].Journal of Modern Laboratory Medicine,2025,40(04):73-78.[doi:10.3969/j.issn.1671-7414.2025.04.013]
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水苏碱调节Hippo-YAP信号通路对高糖诱导心肌细胞损伤模型影响的实验研究()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第40卷
期数:
2025年04期
页码:
73-78
栏目:
论著
出版日期:
2025-07-15

文章信息/Info

Title:
Experimental Study on the Effect of Stachydrine on Myocardial Cell Injury Model Induced by High Glucose Through Regulating Hippo-YAP Signaling Pathway
文章编号:
1671-7414（2025）04-073-06
作者:
宋彦庆，杜章辉，丁冰钰
（山东中医药大学附属医院东营医院内分泌科，山东东营 257000）
Author(s):
SONG Yanqing, DU Zhanghui, DING Bingyu
(Department of Endocrinology, Dongying Hospital, Affiliated Hospital of Shandong University of Chinese Medicine, Shandong Dongying 257000, China)
关键词:
水苏碱; Hippo-Yes 相关蛋白信号通路; 高糖; 心肌细胞; 损伤
分类号:
R-332
DOI:
10.3969/j.issn.1671-7414.2025.04.013
文献标志码:
A
摘要:
目的 研究水苏碱对高糖（HG）诱导的心肌细胞损伤的影响及其与Hippo-Yes 相关蛋白（YAP）信号通路的调节作用。方法 以30 mmol/L 葡萄糖对大鼠心肌细胞H9C2 进行HG 刺激以建立心肌细胞损伤模型，然后用0.05，0.10，0.15，0.20 mmol/L 的水苏碱处理细胞并用CCK-8 法检测其活性，筛选水苏碱的作用浓度。然后将H9C2 细胞分为对照（ctrl）组、HG 组、低浓度水苏碱组、高浓度水苏碱组和高浓度水苏碱+Hippo-YAP 信号通路抑制剂（TDI-011536）组。除ctrl 组用5 mmol/L 葡萄糖培养细胞外，其余4 组均用30 mmol/L 葡萄糖培养，低、高浓度水苏碱组分别用0.05，0.10mmol/L 水苏碱培养24h，高浓度水苏碱+TDI-011536 组用0.10 mmol/L 水苏碱和3.00 μmol/L TDI-011536 培养24h。CCK-8 法检测细胞增殖，流式细胞术检测细胞凋亡，ELISA 法检测丙二醛（MDA）和超氧化物歧化酶（SOD）水平，Westernblot 检测磷酸化YAP（p-YAP），YAP，增殖细胞核抗原（PCNA）和B 淋巴细胞瘤-2 基因（Bcl-2）蛋白水平。结果与ctrl 组比较，0.05，0.10，0.15，0.20 mmol/L 水苏碱处理HG 诱导的H9C2 细胞存活率显著增加，差异具有统计学意义（t=8.32 ～ 29.67，均P<0.01），水苏碱的半数抑制浓度（IC50 值）约为0.09 mmol/L，选择接近和低于IC50 的0.05mmol/L 和0.10 mmol/L 浓度作为后续实验的水苏碱浓度。与ctrl 组比较，HG 组细胞存活率显著降低（t=44.32，11.04，P<0.01）；与HG 组比较，低浓度水苏碱组、高浓度水苏碱组细胞存活率显著升高（t=10.06，21.66，均P<0.01）；与高浓度水苏碱组比较，高浓度水苏碱+TDI-011536 组细胞存活率显著下降（t=9.54，P<0.01），差异具有统计学意义。与ctrl 组比较，HG 组细胞凋亡率显著升高（t=36.74，P<0.01）；与HG 组比较，低浓度水苏碱组、高浓度水苏碱组细胞凋亡率显著下降（t=11.04，26.78，均P<0.01）；与高浓度水苏碱组比较，高浓度水苏碱+TDI-011536 组细胞凋亡率显著升高（t=9.96，均P<0.01），差异具有统计学意义。与ctrl 组比较，HG 组SOD 水平显著降低，MDA 水平显著升高（t=18.85，29.12，均P ＜ 0.01）；与HG 组比较，低浓度水苏碱组、高浓度水苏碱组SOD 水平显著升高（t=6.59，9.86，均P ＜ 0.01），MDA 水平显著降低（t=13.45，23.36，均P ＜ 0.01）；与高浓度水苏碱组比较，高浓度水苏碱+TDI-011536 组SOD 水平显著降低，MDA 水平显著升高（t=5.30，6.98），差异具有统计学意义（均P ＜ 0.01）。与ctrl 组比较，HG组p-YAP，p-YAP/YAP，PCNA，Bcl-2 蛋白水平显著降低，YAP 蛋白水平显著升高（t=15.36~45.00，均P ＜ 0.01）；与HG 组比较，低浓度水苏碱组、高浓度水苏碱组的p-YAP，p-YAP/YAP，PCNA，Bcl-2 蛋白水平显著升高，YAP 蛋白水平显著降低（t=5.51~25.15，均P ＜ 0.01）；与高浓度水苏碱组比较，高浓度水苏碱+TDI-011536 组p-YAP，p-YAP/YAP，PCNA，Bcl-2 蛋白水平显著降低，YAP 蛋白水平显著升高（t=4.27~11.25），差异具有统计学意义（均P ＜ 0.05）。结论 水苏碱可能通过激活Hippo-YAP 信号通路抑制氧化应激和细胞凋亡，进而改善HG 诱导的心肌细胞损伤。
Abstract:
Objective To study the effect of stachydrine on cardiomyocyte damage induced by high glucose (HG) and its regulation of Hippo-Yes-associated protein (YAP) signaling pathway. Methods Rat myocardial cells H9C2 were stimulated by HG with 30 mmol/L glucose to establish the myocardial cell injury model, and then treated with 0.05, 0.10, 0.15, 0.20 mmol/L of stachydrine and their activities were detected by CCK-8 method, and the action concentration of stachycarine was screened. Then H9C2 was grouped into control (ctrl) group, HG group, low concentration stachydrine group, high concentration stachydrine group, and high concentration stachydrine+Hippo-YAP signaling pathway inhibitor (TDI-011536) group. Except for the ctrl group cultured with 5 mmol/L glucose, the other 4 groups were cultured with 30 mmol/L glucose, and the low and high concentration stachydrine groups were cultured with 0.05 and 0.10 mmol/L threonine for 24h, respectively. The high concentration stachydrine +TDI-011536 group were cultured with 0.10 mmol/L stachydrine and 3.00 μmol/L TDI-011536 for 24h. CCK-8 method was applied to detect cell proliferation. Flow cytometry was applied to detect apoptosis. ELISA were applied to detect the level of malondialdehyde (MDA) and superoxide dismutase (SOD). Western blot was applied to detect the level of phosphorylated YAP (p-YAP), YAP, proliferating cell nuclear antigen (PCNA), and B-cell lymphoma-2 (Bcl-2) proteins. Results Compared with ctrl group, the survival rate of H9C2 cells induced by HG was significantly increased by 0.05, 0.10, 0.15, 0.20 mmol/L stachydrine treatment, and the differences were statistically significant (t=8.32~29.67,all P<0.01). The 50% inhibitory concentration (IC50) value of stachydrine was about 0.09 mmol/L, and the concentrations of 0.05 mmol /L and 0.10 mmol /L close to and lower than IC50 were selected as the concentrations of hydrostachyine for subsequent experiments. Compared with the ctrl group, the survival rate in HG group was significantly decreased (t=44.32, P<0.01). Compared with HG group, the cell survival rate of low concentration stachydrine group and high concentration stachydrine group was significantly increased (t=10.06, 21.66, all P<0.01). Compared with the high concentration stachydrine group, the cell survival rate in the highconcentration stachydrine +TDI-011536 group was significantly decreased (t=9.54, P<0.01) , and the differences were statistically significant, respectively. Compared with ctrl group, the apoptosis rate of HG group was significantly increased (t=36.74, P<0.01). Compared with HG group, the apoptosis rate of low concentration stachydrine group and high concentration stachydrine group was significantly decreased (t=11.04, 26.78, all P<0.01). Compared with the high concentration threonine group, the apoptosis rate of the high concentration stachydrine +TDI-011536 group was significantly increased (t=9.96, P<0.01), and the differences were statistically significant, respectively. Compared with ctrl group, SOD level in HG group was significantly decreased, MDA levels were significantly increased (t=18.85, 29.12, all P<0.01). Compared with HG group, SOD level were significantly increased in low concentration stachydrine groups and high concentration stachydrine groups(t=6.59,9.86,all P<0.01), MDA level were significantly decreased (t=13.45, 23.36, all P<0.01). Compared with the high concentration stachydrine group, the SOD level in the high concentration hydrostatin +TDI-011536 group was significantly decreased. MDA levels were significantly increased, and the differences were statistically significant (t=5.30, 6.98, all P<0.01), respectively. Compared with ctrl group,the level of p-YAP, p-YAP/YAP, PCNA, Bcl-2 protein were significantly decreased, and the level of YAP protein was significantly increased (t=15.36 ～ 45.00, all P<0.01). Compared with HG group, the level of p-YAP, p-YAP/YAP, PCNA, Bcl-2 protein were significantly increased in low concentration stachydrine group and high concentration stachydrine group, the level of YAP protein levels were significantly decreased (t=5.51 ～ 25.15, all P<0.01).Compared with the high concentration stachydrine group, the level of p-YAP, p-YAP/YAP, PCNA, Bcl-2 protein in the high concentration hydrothreonine +TDI-011536 group were significantly decreased, the level of YAP protein significantly increased, the differences were statistically significant (t=4.27 ~ 11.25, all P<0.05). Conclusion Stachydrine may inhibit oxidative stress and apoptosis by activating Hippo-YAP signaling pathway, thereby ameliorating HG-induced myocardial cell damage.

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备注/Memo

备注/Memo:
作者简介：宋彦庆（1974-），男，硕士，副主任医师，研究方向：内分泌疾病，E-mail：fdfg_d2022@163.com。
通讯作者：丁冰钰，E-mail：xunxiannaoji303@163.com。
更新日期/Last Update: 2025-07-15