参元益气活血胶囊对肿瘤坏死因子α诱导小鼠心肌细胞损伤的影响及机制

娄妍; 刘红旭; 尚菊菊; 周琦; 周明学; 李享; 邢文龙

doi:10.16025/j.1674-1307.2024.11.011

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参元益气活血胶囊对肿瘤坏死因子α诱导小鼠心肌细胞损伤的影响及机制

实验研究 | 更新时间：2024-12-23

- 参元益气活血胶囊对肿瘤坏死因子α诱导小鼠心肌细胞损伤的影响及机制
- Effect and mechanism of Shenyuan Yiqi Huoxui Capsules on TNF-α-induced myocardial cell injury in mice
- 北京中医药 2024年43卷第11期页码：1275-1280
- 作者机构：
  
  1.首都医科大学附属北京中医医院心血管科，北京 100010
  2.首都医科大学，北京 100069
  3.北京市中医药研究所脑心同治实验室，北京 100010
- 作者简介：
  
  娄妍，女，29岁，博士研究生。研究方向：中医内科心血管病。
  刘红旭，E-mail：lhx_@263.net
- 基金信息：
  
  国家自然科学基金资助项目面上课题(8217151495);国家自然科学基金资助项目青年课题(82205098)
- DOI：10.16025/j.1674-1307.2024.11.011
  中图分类号：
- 纸质出版日期：2024-11-25，
  
  收稿日期：2024-03-04，
- 稿件说明：
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娄妍，刘红旭，尚菊菊，等.参元益气活血胶囊对肿瘤坏死因子α诱导小鼠心肌细胞损伤的影响及机制［J］.北京中医药，2024，43（11）：1275-1280.

LOU Yan,LIU Hongxu,SHANG Juju,et al.Effect and mechanism of Shenyuan Yiqi Huoxui Capsules on TNF-α-induced myocardial cell injury in mice[J]. Beijing Journal of Traditional Chinese Medicine,2024,43(11):1275-1280.
娄妍，刘红旭，尚菊菊，等.参元益气活血胶囊对肿瘤坏死因子α诱导小鼠心肌细胞损伤的影响及机制［J］.北京中医药，2024，43（11）：1275-1280. DOI： 10.16025/j.1674-1307.2024.11.011.

LOU Yan,LIU Hongxu,SHANG Juju,et al.Effect and mechanism of Shenyuan Yiqi Huoxui Capsules on TNF-α-induced myocardial cell injury in mice[J]. Beijing Journal of Traditional Chinese Medicine,2024,43(11):1275-1280. DOI： 10.16025/j.1674-1307.2024.11.011.

摘要

目的

探讨参元益气活血胶囊（SYYQ）对肿瘤坏死因子α（TNF-α）诱导的小鼠心肌细胞损伤的影响及机制。

方法

常规培养小鼠心肌细胞（HL-1细胞），并制备SYYQ含药血清。用0、10%、20%、50%的SYYQ含药血清干预HL-1细胞，用细胞计数试剂法（CCK-8）检测细胞存活率，根据细胞存活率筛选最佳实验浓度。将HL-1细胞随机分为3组，对照组加入DMEM培养，模型组用50 ng/mL的TNF-α刺激12 h诱导心肌损伤模型，SYYQ组在模型组基础上加入最佳实验浓度的含药血清，同步培养12 h。流式细胞术检测细胞凋亡，Real-time PCR检测细胞内核因子κB（NF-κB）、核苷酸结合寡聚域样受体蛋白3（NLRP3）、白细胞介素18（IL-18）mRNA表达，Western blotting法检测细胞内NF-κB、NLRP3、IL-18蛋白表达。

结果

不同浓度SYYQ含药血清干预的心肌细胞存活率分别为（53.32±5.70）%、（68.24±6.41）%、（92.62±8.56）%、（86.93±7.12）%，故将20%含药血清组作为药物干预细胞的最佳浓度。对照组、模型组、SYYQ组心肌细胞凋亡率分别为（1.57±0.18）%、（18.99±1.32）%、（6.76±0.38）%。与对照组比较，模型组心肌细胞凋亡率高（

0.01）；与模型组比较，SYYQ组心肌细胞凋亡率低（

0.01）。与对照组比较，模型组细胞内NF-κB、NLRP3、IL-18 mRNA相对表达量高（

0.05，

0.01）；与模型组比较，SYYQ组细胞内NF-κB、NLRP3、IL-18 mRNA相对表达量低（

0.01）。与对照组比较，模型组细胞内NLRP3、IL-18蛋白相对表达量高（

0.01）；与模型组比较，SYYQ组细胞内NF-κB、NLRP3、IL-18蛋白相对表达量低（

0.05，

0.01）。

结论

SYYQ可能通过调控NF-κB/NLRP3/IL-18信号通路减轻TNF-α所致心肌细胞损伤。

Abstract

Objective

To investigate the effect of Shenyuan Yiqi Huoxue Capsules （SYYQ） on tumor necrosis factor α （TNF-α）-induced myocardial cell injury in mice and its mechanism.

Methods

Mouse cardiomyocytes （HL-1 cells） were cultured routinely， and SYYQ-containing serum was prepared. HL-1 cells were treated with 0%， 10%， 20%， and 50% SYYQ-containing serum. Cell viability was measured using the cell counting kit-8 （CCK-8） assay， and the optimal experimental concentration was selected based on the cell survival rate. HL-1 cells were randomly divided into three groups： the control group， which was cultured with DMEM； the model group， which was stimulated with 50 ng/mL TNF-α for 12 h to induce myocardial injury； the SYYQ group， which was treated with the optimal concentration of drug-containing serum for 12 h. Flow cytometry was used to detect apoptosis， and real-time PCR was used to measure the mRNA expression levels of nuclear factor kappa B （NF-κB）， nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 （NLRP3）， and interleukin-18 （IL-18）. Western blotting was used to detect the protein expression levels of NF-κB， NLRP3， and IL-18.

Results

The survival rates of myocardial cells were （53.32±5.70）%，（68.24±6.41）%，（92.62±8.56）%， and （86.93±7.12）%， respectively. The 20% drug-containing serum group was selected as the optimal concentration for drug intervention. The apoptosis rates of cardiomyocytes in the control group， model group， and SYYQ group were（1.57±0.18）%，（18.99±1.32）%，and（6.76±0.38）%， respectively. Compared with the control group， the apoptosis rate in the model group was significantly higher （

0.01）. Compared with the model group， the apoptosis rate in the SYYQ group was significantly lower （

0.01）. The mRNA expression levels of NF-κB， NLRP3， and IL-18 were significantly higher in the model group compared with the control group （

0.05）. Compared with the model group， the mRNA expression levels of NF-κB， NLRP3， and IL-18 were significantly lower in the SYYQ group （

0.01）. The relative protein expression levels of NLRP3 and IL-18 were significantly higher in the model group compared with the control group （

0.01）. Compared with the model group， the relative protein levels of NF-κB， NLRP3， and IL-18 were significantly lower in the SYYQ group （

0.05，

0.01）.

Conclusion

SYYQ may reduce TNF-α-induced myocardial cell injury by regulating the NF-κB/NLRP3/IL-18 signaling pathway.

关键词

参元益气活血胶囊心肌细胞肿瘤坏死因子α炎症损伤核因子κB核苷酸结合寡聚域样受体蛋白3白细胞介素18小鼠

Keywords

Shenyuan Yiqi Huoxue Capsulescardiomyocytestumor necrosis factor αinflammatory injurynuclear factor κbnucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3interleukin 18mouse

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