脉管Ⅱ号胶囊对大鼠后肢缺血后血管再生的影响及其机制

doi:10.13481/j.1671-587X.20210216

摘要/Abstract

摘要： 目的

探索脉管Ⅱ号胶囊对大鼠肢体缺血症的影响，为外周动脉疾病的治疗奠定理论基础。

方法

60只SPF级雌性SD大鼠随机分为正常对照组、模型组、假手术组、阳性对照组（0.202 5 g·kg^－1复方丹参）、低剂量脉管Ⅱ号胶囊组（30 mg·kg^－1）和高剂量脉管Ⅱ号胶囊组（90 mg·kg^－1），每组10只。采用结扎股动静脉近端、远端及其主要分支的方法建立大鼠后肢缺血模型。观察各组大鼠的一般状态，采用HE染色法观察各组大鼠后肢肌肉组织病理形态表现，采用免疫组织化学法检测各组大鼠肌肉组织中血管生成素1（Ang-1）、酪氨酸激酶受体2（Tie2）和CD31阳性表达细胞数；采用Western blotting法检测各组大鼠肌肉组织中Ang-1和Tie2蛋白表达水平。

结果

大鼠手术侧后肢皮肤温度偏低，皮肤颜色呈现暗红色，后肢体出现紫绀和趾甲脱落等现象。与正常对照组和假手术组比较，模型组大鼠后肢肌肉细胞肿胀明显，出现横纹消失以及肌束膜内肌原纤维萎缩的现象；与模型组比较，阳性对照组、低和高剂量脉管Ⅱ号胶囊组上述情况有所缓解。与假手术组比较，模型组大鼠肌肉组织中Ang-1和Tie2阳性表达细胞数明显减少（P<0.05）；与模型组比较，阳性对照组和高剂量脉管Ⅱ号组大鼠肌肉组织中Ang-1和Tie2阳性表达细胞数明显升高（P<0.05）。与模型组比较，阳性对照组和高剂量脉管Ⅱ号胶囊组大鼠后肢皮肤温度差明显减小（P<0.05），大鼠肌肉组织中Ang-1和Tie2蛋白表达水平均明显升高（P<0.05）。

结论

脉管Ⅱ号胶囊可促进大鼠肢体缺血后血管的再生，其作用机制可能与激活Ang-1/Tie2通路有关。

关键词: 脉管Ⅱ号胶囊, 肢体缺血, 血管生成素1, 酪氨酸激酶受体2, 血管新生

Abstract: Objective

To explore the effect of Vascular Capsule Ⅱ on the limb ischemia of the rats， and to lay a theoretical foundation for the treatment of peripheral arteriopathy.

Methods

A total of 60 SPF female SD rats were randomly divided into normal control group， model group， sham operation group， positive control group （0.202 5 g·kg^－1 Compound Danshen Tablets）， low dose of Vascular Capsule Ⅱ group （30 mg·kg^－1） and high dose of Vascular Capsule Ⅱ group （90 mg·kg^－1）；there were 10 rats in each group. The models of hind limb ischemia were established by ligating the proximal， distal and main branches of femoral artery and vein. The general states of rats were observed. The pathomorphology of hind limb muscle tissue of the rats in various groups was observed by HE staining. The number of cells with positive expression of angiopoietin 1 （Ang-1）， tyrosine kinase receptor 2 （Tie2）， and CD31 in muscle tissue of the rats in various groups were detected by immunohistochemistry. The expression levels of Ang-1 and Tie2 proteins in muscle tissue of the rats in various groups were detected by Western blotting method.

Results

The skin temperature of the hind limbs of the operation rats was low， the skin color was dark red， and cyanosis and toenail dropping occurred in the hind limbs. Compared with normal group and sham operation group， the hind limb muscle cells of the rats in model group were obviously swollen， the transverse striae disappeared， and the myofibril atrophy in the perineurium was observed；compared with model group，the symptoms mentioned above in positive control group and low and high doses of Vascular Capsule Ⅱ groups were alleviated.Compared with sham operation group，the number of cells with positive expressions of Ang-1 and Tie2 in muscle tissue of the rats in model group was significantly decreased（P<0.05）；compared with model group，the number of cells with positive expressions of Ang-1 and Tie2 in muscle tissue of the rats in positive control group and low and high doses of Vascular Capsule Ⅱgroups were significantly increases（P<0.05）. Compared with model group， the skin temperature differences of hind limbs of the rats in positive control and high dose of Vascular Capsule Ⅱ group were significantly decreased （P<0.05）， and the expression levels of CD31， Ang-1， and Tie2 proteins were significantly increased （P<0.05）.

Conclusion

Vascular Capsule Ⅱ can promote the vascular regeneration after limb ischemia in the rats，and its mechanism may be related to activating the Ang-1/Tie2 pathway.

Key words: Vascular Capsule Ⅱ, limb ischemia, angiopoietin-1, tyrosine kinase receptors 2, vascular regeneration

中图分类号:

R285.5

王世军,杨尾莲,陈福伟,张美吉,杨旭. 脉管Ⅱ号胶囊对大鼠后肢缺血后血管再生的影响及其机制[J]. 吉林大学学报(医学版), 2021, 47(2): 369-376.

Shijun WANG,Weilian YANG,Fuwei CHEN,Meiji ZHANG,Xu YANG. Effect of Vascular Capsule Ⅱ on vascular regeneration after hind limb ischemia in rats and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2021, 47(2): 369-376.

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Group	Number of positive cells
Group	Ang-1	Tie2
Normal control	0.36±0.04	0.33±0.01
Sham operation	0.36±0.04	0.31±0.01
Model	0.25±0.02^*	0.23±0.02^*
Positive control	0.32±0.02^△	0.30±0.03^△
Low dose of Vascular CaspuleⅡ	0.25±0.04	0.28±0.02
High dose of Vascular CaspuleⅡ	0.30±0.02^△	0.29±0.02^△

Group	A value
Group	Ang-1	Tie2
Normal control	1.00±0.10	1.00±0.05
Sham operation	0.96±0.14	0.94±0.05
Model	0.39±0.11^*	0.38±0.05^*
Positive control	0.75±0.09^△	0.78±0.06^△
Low dose of Vascular CaspuleⅡ	0.51±0.06^△	0.53±0.09^△
High dose of Vascular CaspuleⅡ	0.72±0.09^△	0.73±0.08^△

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