1.湖南中医药大学第一附属医院,湖南省长沙市雨花区韶山中路95号,410007
2.湖南中医药大学
3.湖南中医药高等专科学校附属第一医院
pqh410007@126.com,(0731)88458010
收稿:2022-07-22,
修回:2022-10-30,
纸质出版:2023-01-17
移动端阅览
易琼,彭清华,郭志华等.心痛泰对痰瘀互结证腹主动脉易损斑块模型兔病理性血管新生及HIF-1α/VEGF/VEGFR2信号通路的影响[J].中医杂志,2023,64(02):159-166.
YI Qiong,PENG Qinghua,GUO Zhihua,et al.Effects of Xintongtai (心痛泰) on Pathological Angiogenesis and HIF-1α/VEGF/VEGFR2 Signaling Pathway in Abdominal Aortic Vulnerable Plaque Model Rabbits with Binding of Phlegm and Stasis Syndrome[J].Journal of Traditional Chinese Medicine,2023,64(02):159-166.
易琼,彭清华,郭志华等.心痛泰对痰瘀互结证腹主动脉易损斑块模型兔病理性血管新生及HIF-1α/VEGF/VEGFR2信号通路的影响[J].中医杂志,2023,64(02):159-166. DOI: 10.13288/j.11-2166/r.2023.02.010.
YI Qiong,PENG Qinghua,GUO Zhihua,et al.Effects of Xintongtai (心痛泰) on Pathological Angiogenesis and HIF-1α/VEGF/VEGFR2 Signaling Pathway in Abdominal Aortic Vulnerable Plaque Model Rabbits with Binding of Phlegm and Stasis Syndrome[J].Journal of Traditional Chinese Medicine,2023,64(02):159-166. DOI: 10.13288/j.11-2166/r.2023.02.010.
目的
2
探讨心痛泰治疗动脉粥样硬化痰瘀互结证的可能作用机制。
方法
2
将60只清洁级兔随机分为假手术组、模型组、氟伐他汀组及心痛泰低、中、高剂量组,每组10只。除假手术组外,其余各组兔采用高脂喂养+腹主动脉球囊损伤法建立痰瘀互结证腹主动脉易损斑块模型。造模后心痛泰低、中、高剂量组分别给予心痛泰含生药1.87、3.73、7.46 g/(kg·d)灌胃,氟伐他汀组给予氟伐他汀钠缓释片1.87 mg/(kg·d)灌胃,假手术组和模型组给予10 ml/d超纯水灌胃,各组均连续灌胃8周。采用HE染色观察易损斑块中病理性血管新生的变化;采用痰瘀互结证中医证候评分表进行证候评分并计算总积分;测定斑块成分(包括细胞外脂质、泡沫细胞、胶原成分、平滑肌细胞各占所在斑块面积的百分比),并计算斑块易损指数;免疫组化法测定斑块组织中缺氧诱导因子1α(HIF-1α)、血管内皮生长因子(VEGF)、血管内皮生长因子受体2(VEGFR2)、血管生成素2(Ang-2)、成纤维细胞生长因子(FGF)阳性表达;蛋白免疫印迹法测定斑块组织中HIF-1α、VEGF、VEGFR2、Ang-2、FGF蛋白表达。
结果
2
HE染色显示,假手术组腹主动脉无斑块形成;模型组可见动脉弹力层破坏,形成易损斑块,内膜下有明显的泡沫细胞,斑块纤维帽破裂;与模型组相比,心痛泰各剂量组和氟伐他汀组斑块组织处的空泡化明显减轻,动脉内膜和斑块的结构更加稳定。与模型组比较,心痛泰各剂量组和氟伐他汀组各证候评分及总积分下降,细胞外脂质含量、泡沫细胞含量降低,胶原含量、平滑肌细胞含量增加,斑块易损指数降低,斑块组织中HIF-1α、VEGF、VEGFR2、Ang-2、FGF阳性表达及蛋白表达降低(
P
<0.05或
P
<0.01);且心痛泰中、高剂量组和氟伐他汀组各指标改善优于心痛泰低剂量组(
P
<0.01)。心痛泰中、高剂量组和氟伐他汀组组间两两比较,各指标差异无统计学意义(
P
>0.05)。
结论
2
心痛泰可能通过下调斑块组织HIF-1α/VEGF/VEGFR2信号通路,减少病理性血管新生,稳定易损斑块,从而改善动脉粥样硬化之痰瘀互结证。
Objective
2
To explore the possible mechanism of
Xintongtai
(心痛泰) in treating atherosclerosis with binding of phlegm and stasis syndrome.
Methods
2
Sixty clean rabbits were randomly divided into sham operation group, model group, fluvastatin group and
Xintongtai
low-, medium- and high-dose groups, with 10 rabbits in each group. Except for the sham operation group, rabbits were given high-fat feeding plus balloon injury of abdominal aorta to establish the vulnerable plaque model of abdominal aorta with binding of phlegm and stasis syndrome. After modelling,
Xintongtai
low-, medium-, and high-dose groups were given
Xintongtai
containing 1.87, 3.73, and 7.46 g/(kg·d) of crude drug by gavage, while the fluvastatin group was given 1.87 mg/(kg·d) of fluvastatin sodium sustained-release tablets by gavage, and the sham operation group and the model group were given intragastric administration of 10 ml/d of ultrapure water, all for 8 weeks. HE staining was used to observe the changes of pathological angiogenesis in vulnerable plaques. The traditional Chinese medicine(TCM) syndrome scale for binding of phlegm and stasis syndrome was used to evaluate the syndromes and calculate the total score. The plaque components including the percentage of extracellular lipids, foam cells, collagen components and smooth muscle cells in the plaque area were measured, and the plaque vulnerability index was calculated. Immunohistochemical method was used to detect the positive expressions of hypoxia-inducible factor 1α (HIF-1α), vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 2 (VEGFR2), angiopoietin 2 (Ang-2), and fibroblast growth factor (FGF) in the plaque tissue. Western blotting was used to detect HIF-1α, VEGF, VEGFR2, Ang-2, FGF protein expression.
Results
2
HE staining showed that there was no plaque in the abdominal aorta in the sham operation group; in the model group, the elastic layer of the arteries was damaged, forming vulnerable plaques, and there were obvious foam cells under the intima, and the fibrous cap of the plaques was ruptured. Compared to those in the model group, the vacuolization of plaque tissue in all
Xintongtai
groups and fluvastatin group was significantly reduced, and the structure of arterial intima and plaque was more stable. All
Xintongtai
groups and the fluvastatin group had lower separate and total TCM scores, decreased extracellular lipids and foam cells, increased collagen components and smooth muscle cells in the plaque area, decreased plague vulnerability index, and lower positive expression and protein expression of HIF-1α, VEGF, VEGFR2, Ang-2, and FGF in plaque tissue than the model group (
P
<0.05 or
P
<0.01); and more improvements were found in the medium- and high-dose
Xintongtai
groups and the fluvastatin group than the low-dose
Xintongtai
group (
P
<0.01). There was no statistically significant differences in all indicators between any two of the
Xintongtai
medium-, high-dose groups and fluvastatin group (
P
>0.05).
Conclusion
2
Xintongtai
may reduce pathological angiogenesis and stabilize vulnerable plaques by down-regulating the HIF-1α/VEGF/VEGFR2 signaling pathway in plaque tissue, thereby improving the binding of phlegm and stasis syndrome in atherosclerosis.
中国心血管健康与疾病报告编写组 . 中国心血管健康与疾病报告2021概要 [J]. 中国循环杂志 , 2022 , 37 ( 6 ): 553 - 578 .
徐斌 , 石祥恩 , 李敏 , 等 . 乙酰肝素酶及新生血管在人颈动脉粥样硬化斑块中的表达 [J]. 中国脑血管病杂志 , 2019 , 16 ( 3 ): 116 - 122 .
陈明真 , 龚慕云 , 傅蓉 , 等 . 滋养血管新生与动脉粥样硬化斑块稳定性的关系 [J]. 世界最新医学信息文摘 , 2019 , 19 ( 43 ): 62 - 65 .
王健 . 化瘀祛痰方调控HIF-1α/VEGF/VEGFR-2通路对AS家兔主动脉斑块的影响及机制研究 [D]. 沈阳 : 辽宁中医药大学 , 2020 .
刘悦 , 张帆 , 王洋 , 等 . 基于“心受气于脾”理论探讨冠心病的发病机制 [J]. 时珍国医国药 , 2019 , 30 ( 7 ): 1675 - 1677 .
李佳玲 , 魏佳明 , 李玉莹 , 等 . 心痛泰颗粒治疗气滞血瘀证稳定型心绞痛的效果及对心肌能量代谢的影响 [J]. 中医药导报 , 2021 , 18 ( 11 ): 62 - 65 .
易琼 , 李雅 , 郭志华 , 等 . 基于TLR4/p38MAPK/AP-1信号通路探讨心痛泰对兔动脉粥样硬化和血管重构的机制 [J]. 中华中医药杂志 , 2020 , 35 ( 8 ): 3878 - 3884 .
易琼 , 李雅 , 郭志华 , 等 . 心痛泰对动脉粥样硬化兔血脂及主动脉ox-LDL、IL-1β的影响 [J]. 中国动脉硬化杂志 , 2016 , 24 ( 1 ): 39 - 43 .
易琼 , 李雅 , 郭志华 , 等 . 心痛泰通过降低 Lp-PLA2、IL-6、hs-CRP、LOX-1 水平减轻兔动脉粥样硬化炎症反应 [J]. 中国动脉硬化杂志 , 2017 , 25 ( 8 ): 796 - 799 .
易琼 , 李雅 , 郭志华 , 等 . 基于“心受气于脾”探讨心痛泰调控p38 MAPK/AP-1对动脉粥样硬化兔的中医证候和VSMC胶原纤维的影响 [J]. 中国实验方剂学杂志 , 2021 , 27 ( 19 ): 56 - 65 .
孙敬方 . 动物实验方法学 [M]. 北京 : 人民卫生出版社 , 2001 : 87 .
张艳 , 杨颖 , 刘锋 , 等 . 高脂饮食联合不同类型球囊损伤制备兔腹主动脉粥样硬化斑块模型实验研究 [J]. 中华实用诊断与治疗杂志 , 2017 , 31 ( 9 ): 850 - 853 .
徐淑云 , 卞如濂 , 陈修 . 药理实验方法学 [M]. 3 版, 北京 : 人民卫生出版社 ,2002: 203 - 206 .
CAMARÉ C , PUCELLE M , NÈGRE-SALVAYRE A , et al . Angiogenesis in the atherosclerotic plaque [J]. Redox Biology , 2017 , 12 : 18 - 34 . doi: 10.1016/j.redox. 2017. 01.007 http://dx.doi.org/10.1016/j.redox.2017.01.007 .
齐炳才 , 靳琦文 , 胡杰 , 等 . 颈动脉粥样硬化斑块内新生血管的研究现状及进展 [J]. 中国动脉硬化杂志 , 2021 , 29 ( 4 ): 359 - 362,368 .
漆仲文 , 李萌 , 朱科 , 等 . 中医药干预动脉粥样硬化易损斑块的机制研究述评 [J]. 中国中医基础医学杂志 , 2021 , 27 ( 1 ): 166 - 169 .
HIGASHIDA T , KANNO H , NAKANO M , et al . Expression of hypoxia-inducible angiogenic proteins (hypoxia-inducible factor-1alpha, vascular endothelial growth factor, and E26 transformation-specific-1) and plaque hemorrhage in human carotid atherosclerosis [J]. J Neurosurg , 2008 , 109 ( 1 ): 83 - 91 .
MAO Y , LIU X , SONG Y , et al . VEGF-A/VEGFR-2 and FGF-2/FGFR-1 but not PDGF-BB/PDGFR-β play important roles in promoting immature and inflammatory intraplaque angiogenesis [J]. PLoS One , 2018 , 13 ( 8 ): e0201395 .
PEACH CJ , MIGNONE VW , ARRUDA MA , et al . Mole-cular pharmacology of VEGF-A isoform:binding and signaling at VEGFR2 [J]. Int J Mol Sci , 2018 , 19 ( 4 ): 1264 .
VRIES MR , PARMA L , PETERS HA , et al . Blockade of vascular endothelial growth factor receptor 2 inhibits intraplaque haemorrhage by normalization of plaque neovessels [J]. J Int Med , 2019 , 285 ( 1 ): 59 - 74 .
KWAK HJ , SO JN , LEE SJ , et al . Angiopoietin-1 is an apoptosis survival factor for endothelial cells [J]. FEBS Lett , 1999 , 448 ( 2 ): 249 - 253 .
HASHIZUME H , FALCON BL , KURODA T , et al . Complementary actions of inhibitors of angiopoietin-2 and VEGF on tumor angiogenesis and growth [J]. Cancer Res , 2010 , 70 ( 6 ): 2213 - 2223 .
GOEDE V , SCHMIDT T , KIMMINA S , et al . Analysis of blood vessel maturation processes during cyclic ovarian angiogenesis [J]. Lab Invest , 1998 , 78 ( 11 ): 1385 - 1394 .
0
浏览量
6
下载量
6
CSCD
关联资源
相关文章
相关作者
相关机构
京公网安备11010102007558号