首页|补阳还五汤激活PDGF信号通路促进小鼠后肢缺血后动脉生成的作用机制

补阳还五汤激活PDGF信号通路促进小鼠后肢缺血后动脉生成的作用机制

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基于血小板衍生生长因子(platelet-derived growth factor,PDGF)信号通路探讨补阳还五汤对缺血组织血流恢复和动脉生成的影响及机制。40 只C57BL/6 小鼠随机分为模型组、阳性对照药贝前列素钠组和补阳还五汤低、中、高剂量组,每组 8只。行股动脉结扎建立后肢缺血模型,模型组给予纯净水 10 mL·kg-1·d-1灌胃,贝前列素钠组给予贝前列素钠 18 μg·kg-1·d-1灌胃,补阳还五汤低、中、高剂量组分别给予补阳还五汤 10、20、40 g·kg-1·d-1灌胃,连续 14 d。采用激光散斑血流灌注成像仪观察小鼠缺血后肢血液灌注,缺血腓肠肌冰冻切片染CD31、α-SMA抗体标记毛细血管和小动脉,蛋白免疫印迹法检测缺血腓肠肌PDGFB和PDGF下游信号分子(p-MEK、MEK、p-ERK、ERK)的表达,real-time PCR检测缺血腓肠肌PDGFB mRNA表达。小鼠原代血管平滑肌细胞(vascular smooth muscle cells,VSMCs)低氧培养建立缺氧模型,按体积比分别加入10%和20%补阳还五汤含药血清,应用增殖、迁移实验观察VSMCs促动脉生成的功能。结果显示,与模型组比较,贝前列素钠组和补阳还五汤低、中、高剂量组小鼠后肢缺血后血流恢复增加,毛细血管、小动脉生成增多,其中补阳还五汤中剂量组效果最显著,PDGFB、p-MEK、p-ERK蛋白及PDGFB mRNA表达明显升高;10%补阳还五汤含药血清处理的VSMCs增殖、迁移功能显著增强。结果表明,补阳还五汤促进缺血后PDGFB转录,激活PDGF信号通路(p-MEK/p-ERK表达升高),进而推动VSMCs增殖、迁移形成小动脉,从而恢复血液灌注,发挥保护缺血组织的作用。
Buyang Huanwu Decoction promotes arteriogenesis after hindlimb ischemia in mice by activating PDGF signaling pathway
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 em-ployed 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 pro-liferation and migration of VSMCs.Our findings demonstrate that Buyang Huanwu Decoction up-regulates PDGFB transcription and ac-tivates PDGF signaling pathway to promote arteriogenesis and blood flow recovery in ischemic gastrocnemius.

ischemiaBuyang Huanwu Decoctionarteriogenesisvascular smooth muscle cellsPDGF signaling pathway

窦永青、马涛、李志杰、马秀茹、张翠娟、郭倩楠、王梦阳、陈建

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河北中医药大学, 河北 石家庄 050200

石家庄市第二医院, 河北 石家庄 050051

缺血 补阳还五汤 动脉生成 血管平滑肌细胞 PDGF信号通路

河北省自然科学基金河北省医学科学研究项目河北省中医药类科学研究课题

H2021423047202201832024093

2024

中国中药杂志
中国药学会

中国中药杂志

CSTPCD北大核心
影响因子:1.718
ISSN:1001-5302
年,卷(期):2024.49(1)
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