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补阳还五汤通过激活血小板衍生生长因子信号通路促进小鼠后肢缺血后的动脉生成

[Buyang Huanwu Decoction promotes arteriogenesis after hindlimb ischemia in mice by activating PDGF signaling pathway].

作者信息

Dou Yong-Qing, Ma Tao, Li Zhi-Jie, Ma Xiu-Ru, Zhang Cui-Juan, Guo Qian-Nan, Wang Meng-Yang, Chen Jian

机构信息

Hebei University of Chinese Medicine Shijiazhuang 050200, China.

the Second Hospital of Shijiazhuang Shijiazhuang 050051, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2024 Jan;49(1):216-223. doi: 10.19540/j.cnki.cjcmm.20230919.402.

Abstract

This study aims to investigate the effect of Buyang Huanwu Decoction on blood flow recovery and arteriogenesis after hindlimb ischemia in mice via the platelet-derived growth factor(PDGF) signaling pathway. Forty C57BL/6 mice were randomized into model(clean water, 10 mL·kg(-1)·d(-1)), beraprost sodium(positive control, 18 μg·kg(-1)·d(-1)), and low-, medium-, and high-dose(10, 20, and 40 g·kg(-1)·d(-1), respectively) Buyang Huanwu Decoction groups(n=8). The hindlimb ischemia model was established by femoral artery ligation. The mice were administrated with corresponding agents by gavage daily for 14 days after ligation. For laser Doppler perfusion imaging, the mice were anesthetized and measured under a Periscan PSI imager. The density of capillary and arterio-le in the ischemic gastrocnemius was measured using immunofluorescence staining of the frozen tissue sections. Western blot was employed to determine the expression of PDGF subunit B(PDGFB), phosphorylated mitogen extracellular kinase(p-MEK), MEK, phosphorylated extracellular signal-regulated kinase(p-ERK), and ERK. Real-time PCR was employed to determine the mRNA level of PDGFB. The Buyang Huanwu Decoction-containing serum was used to treat the vascular smooth muscle cells(VSMCs) in hypoxia at doses of 10% and 20%. The proliferation and migration of VSMCs was assessed in vitro. The results showed that compared with the model group, beraprost sodium and Buyang Huanwu Decoction enhanced the blood flow recovery, increased the capillary and arteriole density, and up-regulated the protein levels of PDGFB, p-MEK, p-ERK, and mRNA levels of PDGFB, with the medium-dose Buyang Huanwu Decoction demonstrating the most significant effect. The 10% Buyang Huanwu Decoction-containing serum enhanced the proliferation and migration of VSMCs. Our findings demonstrate that Buyang Huanwu Decoction up-regulates PDGFB transcription and activates PDGF signaling pathway to promote arteriogenesis and blood flow recovery in ischemic gastrocnemius.

摘要

本研究旨在通过血小板衍生生长因子(PDGF)信号通路,探讨补阳还五汤对小鼠后肢缺血后血流恢复和动脉生成的影响。将40只C57BL/6小鼠随机分为模型组(清水,10 mL·kg⁻¹·d⁻¹)、贝前列素钠组(阳性对照,18 μg·kg⁻¹·d⁻¹)以及低、中、高剂量补阳还五汤组(分别为10、20和40 g·kg⁻¹·d⁻¹,n = 8)。通过股动脉结扎建立后肢缺血模型。结扎后每天经口灌胃给予小鼠相应药物,持续14天。对于激光多普勒灌注成像,将小鼠麻醉后在Periscan PSI成像仪下进行测量。使用冷冻组织切片的免疫荧光染色法测量缺血腓肠肌中毛细血管和微动脉的密度。采用蛋白质印迹法测定PDGF亚基B(PDGFB)、磷酸化丝裂原细胞外激酶(p-MEK)、MEK、磷酸化细胞外信号调节激酶(p-ERK)和ERK的表达。采用实时定量PCR测定PDGFB的mRNA水平。用含10%和20%补阳还五汤的血清处理缺氧状态下的血管平滑肌细胞(VSMCs),在体外评估VSMCs的增殖和迁移情况。结果显示,与模型组相比,贝前列素钠和补阳还五汤可促进血流恢复,增加毛细血管和微动脉密度,并上调PDGFB、p-MEK、p-ERK的蛋白水平以及PDGFB的mRNA水平,其中中剂量补阳还五汤的效果最为显著。含10%补阳还五汤的血清可增强VSMCs的增殖和迁移。我们的研究结果表明,补阳还五汤上调PDGFB转录并激活PDGF信号通路,从而促进缺血腓肠肌的动脉生成和血流恢复。

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