针刺研究

2021, v.46(09) 751-756

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针刺促进血管新生延长脑梗死溶栓时间窗的研究
Acupuncture intervention prolongates thrombolysis time window by regulating cerebral cortex angiogenesis-related genes and proteins in cerebral infarction rats

常思琦;张新昌;张奥;宋扬扬;姜思媛;张智慧;蔡筝韵;刘芙蓉;倪光夏;
CHANG Si-qi;ZHANG Xin-chang;ZHANG Ao;SONG Yang-yang;JIANG Si-yuan;ZHANG Zhi-hui;CAI Zheng-yun;LIU Fu-rong;NI Guang-xia;College of Acupuncture and Massage, Nanjing University of Chinese Medicine;

摘要(Abstract):

目的:通过观察针刺对脑梗死大鼠溶栓治疗后血管新生相关指标表达的影响,探讨针刺延长脑梗死溶栓时间窗的机制。方法:将48只SD大鼠随机分为假手术组、模型组、溶栓组、针刺+溶栓组,每组12只。采用改良自体血栓法制备脑梗死大鼠模型。溶栓组在造模后6 h经尾静脉注射重组人组织型纤溶酶原激活剂(rt-PA,10 mg/kg);针刺+溶栓组于造模后2 h针刺"水沟"、双侧"内关",留针30 min,再于造模后6 h进行溶栓。各组均在造模时进行脑血流量监测;采用Bederson法于造模后2、24 h进行神经行为学评分;氯化三苯基四氮唑(TTC)染色法比较各组大鼠脑梗死体积;Western blot法及荧光定量PCR法检测各组大鼠右侧大脑皮层缺血区血管内皮生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)、内皮抑素(ES)蛋白及mRNA的表达。结果:与假手术组比较,模型组大鼠神经行为学评分、脑梗死体积百分比、大脑皮层缺血区VEGF、bFGF、ES蛋白及mRNA的表达水平明显升高(P<0.01,P<0.05);与模型组比较,针刺+溶栓组神经行为学评分及脑梗死体积百分比降低(P<0.05,P<0.01),大脑皮层缺血区VEGF、bFGF的蛋白及mRNA表达升高(P<0.01,P<0.05),ES蛋白及mRNA表达降低(P<0.01,P<0.05);与溶栓组比较,针刺+溶栓组神经行为学评分及脑梗死体积百分比降低(P<0.05,P<0.01),大脑皮层缺血区VEGF、bFGF蛋白及mRNA表达升高(P<0.01), ES蛋白及mRNA表达降低(P<0.01,P<0.05)。结论:针刺及时介入可延长脑梗死溶栓时间窗,其机制可能与针刺提高缺血大脑皮层VEGF、bFGF蛋白及mRNA表达并抑制ES蛋白及mRNA表达有关。
Objective To observe the effect of acupuncture on the expression of vascular endothelial growth factor(VEGF), basic fibroblast growth factor(bFGF) and endostatin(Endostatin, ES) mRNAs and proteins(angiogenesis related factors) in the ischemic penumbra region in rats with cerebral infarction(CI), so as to explore its underlying mechanisms in prolonging the time window of thrombolysis therapy for CI. Methods A total of 48 male SD rats were randomly divided into sham operation, model, medication(6 h thrombolysis) and acupuncture(Acupunct)+medication groups(n=12 in each group). The CI model was established using modified auto-thrombus method. Six hours after thrombolysis, recombinant human tissue plasminogen activator(rt-PA,10 mg/kg) was given to rats of the thrombolysis group through tail vein. Acupuncture was applied at "Shuigou"(CV26) and bilateral "Neiguan"(PC6) 2 h after successful modeling, and the needles were retained for 30 minutes. Cerebral blood flow(CBF) was monitored during modeling in each group, and the neurological deficit score(0-7 points) was given 2 h and 24 h after successful modeling according to Bederson's methods. The cerebral infarction volume was observed after triphenyltetrazole chloride(TTC) staining. The protein and mRNA expression levels of VEGF, bFGF and ES in the ischemic penumbra region of the right cerebral cortex were detected by Western blot and real-time PCR, separately. Results The neurological deficit score at both 2 h and 24 h after modeling, percentage of cerebral infarction volume, and the expression levels of VEGF, bFGF and ES proteins and mRNAs in the model group were significantly higher than those of the sham operation group(P<0.01, P < 0.05). Compared with the model group, the neurological deficit score 24 h(not at 2 h) after modeling and percentage of cerebral infarction volume, and the expression levels of ES protein and mRNA in the Acupunct+medication group(not in the medication group) were notably lower(P<0.05, P<0.01), while the expression levels of VEGF and bFGF proteins and mRNAs in the Acupunct +medication group(not in the medication group) were considerably higher(P<0.01, P<0.05). No significant differences were found between medication and model groups in the CI percentage, VEGF, bFGF and ES proteins and mRNAs(P>0.05). The therapeutic effect of Acupunct +medication group was significantly superior to that of medication in lowering neurological deficit score, percentage of CI volume and expression of ES protein and mRNA and in up-regulating the expression of VEGF and bFGF proteins and mRNAs(P<0.05, P<0.01). Conclusion Acupuncture and timely intervention can prolong the time window of thrombolysis in CI rats, which may be related to its function in up-regulating the expression of VEGF and bFGF mRNAs and proteins and in down-regulating the expression of ES mRNA and protein in ischemic cerebral cortex.

关键词(KeyWords): 脑梗死;针刺;溶栓时间窗;血管新生
Cerebral infarction;Acupuncture;Thrombolysis time window;Angiogenesis

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(No.81674067);; 江苏省护理学优势学科项目(No.2019YSHL055)

作者(Author): 常思琦;张新昌;张奥;宋扬扬;姜思媛;张智慧;蔡筝韵;刘芙蓉;倪光夏;
CHANG Si-qi;ZHANG Xin-chang;ZHANG Ao;SONG Yang-yang;JIANG Si-yuan;ZHANG Zhi-hui;CAI Zheng-yun;LIU Fu-rong;NI Guang-xia;College of Acupuncture and Massage, Nanjing University of Chinese Medicine;

Email:

DOI: 10.13702/j.1000-0607.201121

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