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

您当前的位置：

首页 >

文章列表页 >

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
- Published：20 February 2023，
  
  Published Online：17 May 2022，
  
  Received：12 January 2022，
- 稿件说明：
扫描看全文
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

芪红通络方深静脉血栓形成血管内皮细胞血小板活化因子核转录因子-κBRas相关的C3肉毒素底物1Ras相关的C3肉毒素底物2补阳还五汤

Keywords

Qihong Tongluo prescriptiondeep venous thrombosisvascular endothelial cellsplatelet-ctivating factornuclear transcription factor- κBRas related C3 botulinum toxin substrate 1Ras related C3 botulinum toxin substrate 2Buyang Huanwutang

references

NICHOLSON M，CHAN N，BHAGIRATH V，et al. Prevention of venous thromboembolism in 2020 and beyond［J］. J Clin Med，2020，9（8）：2467.

KRÜGER-GENGE A，BLOCKI A，FRANKE R P，et al. Vascular endothelial cell biology：An update［J］. Int J Mol Sci，2019，20（18）：4411.

KONG Y，GAO Y，LAN D，et al. Trans-repression of NF-κB pathway mediated by PPARγ improves vascular endothelium insulin resistance［J］. J Cell Mol Med，2019，23（1）：216-226.

KRUEGER I，GREMER L，MANGELS L，et al. Reelin amplifies glycoprotein Ⅵ activation and alphaⅡb beta3 integrin outside-in signaling via PLC gamma 2 and Rho GTPases［J］. Arterioscler Thromb Vasc Biol，2020，40（10）：2391-2403.

KRUGER P C，EIKELBOOM J W，DOUKETIS J D，et al. Deep vein thrombosis：Update on diagnosis and management［J］. Med J Aust，2019，210（11）：516-524.

李彦丽，武如冰，于健，等. 益气活血通脉汤加味对下肢深静脉血栓后综合征（气虚血瘀证）再通的影响及作用机制［J］. 中国实验方剂学杂志，2018，24（3）：202-207.

左敬雅，孙云朝，楚信强，等. “芪红通络方”联合射频消融治疗下肢静脉曲张合并血栓性浅静脉炎40例临床研究［J］. 江苏中医药，2021，53（1）：33-36.

YAO X，CHEN W，LIU J，et al. Deep vein thrombosis is modulated by inflammation regulated via sirtuin 1/NF-κB signalling pathway in a rat model［J］. Thromb Haemost，2019，119（3）：421-430.

黄继汉，黄晓晖，陈志扬，等.药理试验中动物间和动物与人体间的等效剂量换算［J］.中国临床药理学与治疗学，2004，9（9）：1069-1072.

KIRKILESIS G，KAKKOS S K，BICKNELL C，et al. Treatment of distal deep vein thrombosis［J］. Cochrane Database Syst Rev，2020，4（4）：CD013422.

KRÖGER K，MOERCHEL C，THROMBOSE-INITIATIVE E V. Acute deep vein thrombosis-modern diagnostics and treatment［J］. Chirurg，2019，90（1）：71-84.

张婷，楚信强，张欣，等. 芪红通络方联合静脉腔内射频消融术治疗大隐静脉曲张的临床观察［J］. 中医药导报，2020，26（11）：80-83.

马宣，朱运奎. 黄芪提取物对急性肺栓塞兔模型肺组织p38-MAPK通路表达的影响［J］. 世界中西医结合杂志，2021，16（1）：67-72.

WANG Y L，ZHANG Q，YIN S J，et al. Screening of blood-activating active components from Danshen-Honghua herbal pair by spectrum-effect relationship analysis［J］. Phytomedicine，2019，54（1）：149-158.

王丽辉，孙治霞，索红亮，等. 基于HMGB1/TLR4/NF-κB通路探讨桃仁承气汤调节肠源性脓毒症大鼠肠道肌电活动及微环境稳态的作用机制［J］. 中国实验方剂学杂志，2020，26（12）：78-84.

CHENG C Y，KAO S T，LEE Y C. Ferulic acid exerts anti-apoptotic effects against ischemic iInjury by activating HSP70/Bcl-2- and HSP70/autophagy-mediated signaling after permanent focal cerebral ischemia in rats［J］. Am J Chin Med，2019，47（1）：39-61.

刘晓颖. 鸡血藤药用价值及林下栽植应用研究进展［J］. 安徽农业科学，2021，49（20）：28-31，75.

DHAUN N，WEBB D J. Endothelins in cardiovascular biology and therapeutics［J］. Nat Rev Cardiol，2019，16（8）：491-502.

曹亮，刘建华，冯平，等. 黄芪多糖通过TLR4/NF-κB通路抑制COPD大鼠炎性反应诱导的平均血小板体积降低［J］. 疑难病杂志，2020，19（7）：723-729.

OMIDKHODA S F，RAZAVI B M，IMENSHAHIDI M，et al. Evaluation of possible effects of crocin against nitrate tolerance and endothelial dysfunction［J］. Iran J Basic Med Sci，2020，23（3）：303-310.

LI F F，YUAN Y，LIU Y，et al. Pachymic acid protects H9c2 cardiomyocytes from lipopolysaccharide-induced inflammation and apoptosis by inhibiting the extracellular signal-regulated kinase 1/2 and p38 pathways［J］. Mol Med Rep，2015，12（2）：2807-2813.

李春英，易艳，赵雍，等. 香丹注射液和清开灵注射液联合用药对急性炎症血栓模型大鼠脏器的保护作用［J］. 中国实验方剂学杂志，2021，27（23）：80-87.

OU M，ZHANG Y，CUI S，et al. Upregulated miR-9-5p protects against inflammatory response in rats with deep vein thrombosis via inhibition of NF-κB p50［J］. Inflammation，2019，42（6）：1925-1938.

ASLAN J E. Platelet Rho GTPase regulation in physiology and disease［J］. Platelets，2019，30（1）：17-22.

王日生，蔡俊，张继平. 补阳还五汤对大鼠急性脑缺血再灌注损伤的保护作用及机制［J］. 实用医学杂志，2015，31（5）：725-727.

李玲美，付建华，郭浩，等. 注射用红花黄色素经TLR-NF-κB炎症通路抗大鼠MIRI的保护作用研究［J］. 中国中药杂志，2019，44（12）：2566-2571.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Effect of Buyang Huanwutang in Treating Diabetic Peripheral Neuropathy by Inhibiting Pyroptosis Through AMPK/ULK1 Mitophagy Pathway

Effect of Buyang Huanwutang on Skeletal Muscle Injuries in Mouse Model of Type 2 Diabetes Mellitus

Buyang Huanwutang Alleviates Inflammatory Injury After Cerebral Ischemia-reperfusion in Rats via Activating Adiponectin Pathway

Mechanism of Buyang Huanwutang in Regulating Macrophage Cell Polarization Based on TLR4/NF-κB/NLRP3 Pathway

Effect of Buyang Huanwutang on Ferroptosis in Diabetic Kidney Disease Mice

Related Author

AN Jingwen

SONG Linchun

CHEN Die

ZHANG Wang

TIAN Jiaxin

ZHANG Tianya

BEN Ying

WANG Yang

Related Institution

College of Integrated Traditional Chinese and Western Medicine，Hebei University of Chinese Medicine，Hebei Key Laboratory of Integrated Medicine on Liver-kidney Patterns

School of Basic Medicine，Changchun University of Chinese Medicine

School of Basic Medical Sciences，Guangzhou University of Chinese Medicine

The First Affiliated Hospital of Nanchang University

Jiangxi University of Chinese Medicine

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰

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