Protective Effect of Qihong Tongluo Prescription on Vascular Endothelial Cells in Rats with Deep Venous Thrombosis Based on NF-<italic style="font-style: italic">κ</italic>B Pathway

CHU Xinqiang; ZHU Yana; SU Kun; ZHANG Ting; MA Yunlong

doi:10.13422/j.cnki.syfjx.20230495

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Protective Effect of Qihong Tongluo Prescription on Vascular Endothelial Cells in Rats with Deep Venous Thrombosis Based on NF-κB Pathway

更新时间：2023-01-17

- Protective Effect of Qihong Tongluo Prescription on Vascular Endothelial Cells in Rats with Deep Venous Thrombosis Based on NF-κB Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 4, Pages: 60-68(2023)
- 作者机构：
  
  河北省中医院，石家庄 050000
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20230495
  CLC： R22;R242;R2-031;R285.5;R543.6
- Received：12 January 2022，
  
  Published Online：17 May 2022，
  
  Published：20 February 2023
- 稿件说明：
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楚信强,朱雅娜,苏坤等.基于NF-κB通路探讨芪红通络方对深静脉血栓形成大鼠血管内皮细胞的保护作用[J].中国实验方剂学杂志,2023,29(04):60-68.

CHU Xinqiang,ZHU Yana,SU Kun,et al.Protective Effect of Qihong Tongluo Prescription on Vascular Endothelial Cells in Rats with Deep Venous Thrombosis Based on NF-κB Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(04):60-68.
楚信强,朱雅娜,苏坤等.基于NF-κB通路探讨芪红通络方对深静脉血栓形成大鼠血管内皮细胞的保护作用[J].中国实验方剂学杂志,2023,29(04):60-68. DOI： 10.13422/j.cnki.syfjx.20230495.

CHU Xinqiang,ZHU Yana,SU Kun,et al.Protective Effect of Qihong Tongluo Prescription on Vascular Endothelial Cells in Rats with Deep Venous Thrombosis Based on NF-κB Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(04):60-68. DOI： 10.13422/j.cnki.syfjx.20230495.

摘要

目的

探讨芪红通络方对深静脉血栓形成（DVT）大鼠血管内皮细胞的保护作用与机制。

方法

取66只SD大鼠采用随机数字表分为空白组（11只）、建模组（55只），后者通过减慢血流+损伤血管内皮建立DVT模型，并将建模成功大鼠按照随机数字表分为模型组，阿司匹林组，芪红通络方低、中、高剂量组。阿司匹林组予以200 mg·kg

-1

阿司匹林灌胃，芪红通络方低、中、高剂量组分别予以6.5、13、26 g·kg

-1

芪红通络方流浸膏灌胃，模型组和空白组分别予以生理盐水灌胃，每天1次，连续7 d。处死大鼠，取腹主动脉血采用酶联免疫吸附测定法（ELISA）检测血清内皮素-1（ET-1）、白细胞介素-6（IL-6）水平；苏木素-伊红（HE）染色观察血管内皮组织病理改变；透射电镜观察血管内皮细胞超微结构；噻唑蓝（MTT）比色法检测血管内皮细胞活力、乳酸脱氢酶（LDH）试剂盒检测血管内皮细胞LDH释放水平；实时荧光定量聚合酶链式反应（Real-time PCR）检测血管内皮组织血小板活化因子（PAF）、核转录因子-

B（NF-

B）、Ras相关的C3肉毒素底物1（Rac1）、Ras相关的C3肉毒素底物2（Rac2） mRNA表达；蛋白免疫印迹法（Western blot）检测血管内皮组织PAF、NF-

B、Rac1、Rac2蛋白表达。

结果

模型组血管内皮细胞严重损伤并大量脱落，细胞肿胀明显，有大量炎性细胞附着，电镜下可见胞核固缩变形，线粒体高度肿胀，胞质空泡化严重，内弹力膜暴露；芪红通络方各剂量组和阿司匹林组血管内皮组织及其超微结构损伤均有不同程度改善。与空白组比较，模型组血清ET-1、IL-6水平、血管内皮细胞活力、血管内皮组织PAF、NF-

B、Rac1、Rac2 mRNA和蛋白表达明显升高，血管内皮细胞LDH释放水平降低，差异均有统计学意义（

0.05）；与模型组比较，阿司匹林组和芪红通络方各剂量组血清ET-1、IL-6水平、血管内皮细胞活力、血管内皮组织PAF、NF-

B、Rac1、Rac2 mRNA和蛋白表达均降低，血管内皮细胞LDH释放水平升高，差异均有统计学意义（

0.05），其中芪红通络方的作用呈剂量依赖性。

结论

芪红通络方对DVT大鼠血管内皮细胞有保护作用，能防治血栓的形成，推测是通过抑制PAF、NF-

B、Rac1和Rac2的表达，降低血清ET-1、IL-6的水平实现的。

Abstract

Objective

To explore the protective effect and mechanism of Qihong Tongluo prescription on vascular endothelial cells in rats with deep venous thrombosis （DVT）.

Method

Sixty-six SD rats were randomly divided into a blank group （

=11） and a modeling group （

=55）. The DVT model was induced in rats of the modeling group by slowing down blood flow and damaging vascular endothelium. The model rats were randomly divided into model group， aspirin group （200 mg·kg

-1

）， and low-，medium-， and high-dose Qihong Tongluo prescription groups （6.5， 13， 26 g·kg

-1

） according to a random number table. Rats were treated with corresponding drugs by gavage， while those in the model group and the blank group received normal saline， once per day for 7 days. The rats were sacrificed and the abdominal aortic blood was taken. The levels of serum endothelin-1 （ET-1） and interleukin-6 （IL-6） were detected by enzyme-linked immunosorbent assay （ELISA）. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in vascular endothelial tissues. The ultrastructure of vascular endothelial cells was observed by the transmission electron microscope. The viability of vascular endothelial cells was detected by methylthiazolyldiphenyl-tetrazolium bromide （MTT） method，and the release level of lactate dehydrogenase （LDH） was detected by the LDH kit. The messenger ribonucleic acid （mRNA） expression of platelet-activating factor （PAF），nuclear transcription factor

B （NF-

B），Ras-related C3 botulinum toxin substrate 1 （Rac1）， and Ras-related C3 botulinum toxin substrate 2 （Rac2） in vascular endothelial tissues were detected by real-time reverse transcription polymerase chain reaction （Real-time PCR）. The protein expression of PAF，NF-

B，Rac1， and Rac2 in vascular endothelial tissues was detected by Western blot.

Result

The model group showed seriously damaged and swollen vascular endothelial cells with massive shedding， attachment of massive inflammatory cells， nucleus pyknosis and deformation under the electron microscope， highly swollen mitochondria， serious cytoplasmic vacuolation，and exposure of internal elastic membrane. The damage of vascular endothelium and its ultrastructure in Qihong Tongluo prescription groups and the aspirin group was improved in varying degrees. Compared with the blank group，the model group showed increased levels of serum ET-1 and IL-6，potentiated vascular endothelial cell viability， up-regulated mRNA and protein expression of PAF，NF-

B，Rac1， and Rac2 in vascular endothelial tissues，and decreased LDH release level of vascular endothelial cells （

0.05）. Compared with the model group，the aspirin group and the Qihong Tongluo prescription groups showed decreased levels of serum ET-1 and IL-6，blunted vascular endothelial cell viability，down-regulated mRNA and protein expression of PAF，NF-

B，Rac1， and Rac2 in vascular endothelial tissues，and increased LDH release level of vascular endothelial cells （

0.05）. The effect of Qihong Tongluo prescription was dose-dependent.

Conclusion

Qihong Tongluo prescription has a protective effect on vascular endothelial cells of DVT rats and can prevent and treat thrombosis，and its therapeutic effect is presumably achieved by inhibiting the expression of PAF，NF-

B，Rac1，and Rac2 and reducing the levels of serum ET-1 and IL-6.

关键词

Keywords

references

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Protective Effect of Qihong Tongluo Prescription on Vascular Endothelial Cells in Rats with Deep Venous Thrombosis Based on NF-κB Pathway

DOI：10.13422/j.cnki.syfjx.20230495

摘要

Abstract

关键词

Keywords

references