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[针刺干预通过调节脑梗死大鼠大脑皮层血管生成相关基因和蛋白延长溶栓时间窗]

[Acupuncture intervention prolongates thrombolysis time window by regulating cerebral cortex angiogenesis-related genes and proteins in cerebral infarction rats].

作者信息

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

机构信息

College of Acupuncture and Massage, Nanjing University of Chinese Medicine, Nanjing 210023, China.

出版信息

Zhen Ci Yan Jiu. 2021 Sep 25;46(9):751-6. doi: 10.13702/j.1000-0607.201121.

Abstract

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 (<0.01, < 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 (<0.05, <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 (<0.01, <0.05). No significant differences were found between medication and model groups in the CI percentage, VEGF, bFGF and ES proteins and mRNAs (>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 (<0.05, <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.

摘要

目的

观察针刺对脑梗死(CI)大鼠缺血半暗带区血管内皮生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)及内皮抑素(Endostatin,ES)mRNA和蛋白(血管生成相关因子)表达的影响,以探讨其延长CI溶栓治疗时间窗的潜在机制。

方法

将48只雄性SD大鼠随机分为假手术组、模型组、药物治疗(6 h溶栓)组和针刺(Acupunct)+药物治疗组(每组12只)。采用改良自体血栓法建立CI模型。溶栓6 h后,经尾静脉给予溶栓组大鼠重组人组织型纤溶酶原激活剂(rt-PA,10 mg/kg)。造模成功后2 h针刺“水沟”(CV26)及双侧“内关”(PC6),留针30 min。每组在造模过程中监测脑血流量(CBF),并在造模成功后2 h和24 h根据Bederson法给予神经功能缺损评分(0 - 7分)。经氯化三苯基四氮唑(TTC)染色后观察脑梗死体积。分别采用蛋白质印迹法和实时荧光定量PCR检测右侧大脑皮质缺血半暗带区VEGF、bFGF和ES的蛋白及mRNA表达水平。

结果

模型组造模后2 h和24 h的神经功能缺损评分、脑梗死体积百分比以及VEGF、bFGF和ES蛋白及mRNA表达水平均显著高于假手术组(<0.01,<0.05)。与模型组相比,针刺+药物治疗组造模后24 h(而非2 h)的神经功能缺损评分、脑梗死体积百分比以及ES蛋白及mRNA表达水平显著降低(<0.05,<0.01),而针刺+药物治疗组(而非药物治疗组)VEGF和bFGF蛋白及mRNA表达水平显著升高(<0.01,<0.05)。药物治疗组与模型组在CI百分比、VEGF、bFGF和ES蛋白及mRNA方面无显著差异(>0.05)。针刺+药物治疗组在降低神经功能缺损评分、CI体积百分比以及ES蛋白及mRNA表达水平和上调VEGF和bFGF蛋白及mRNA表达水平方面的治疗效果显著优于药物治疗组(<0.05,<0.01)。

结论

针刺及适时干预可延长CI大鼠的溶栓时间窗,这可能与其上调缺血脑皮质VEGF和bFGF mRNA及蛋白表达以及下调ES mRNA和蛋白表达的作用有关。

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