本文選題：頭穴透刺 + 預(yù)處理　； 參考：《黑龍江省中醫(yī)藥科學(xué)院》2016年碩士論文

【摘要】：目的:根據(jù)中醫(yī)“治未病”思想,研究探討頭穴透刺預(yù)處理對(duì)大鼠腦缺血再灌注后內(nèi)源性NSCs的增殖、分化以及對(duì)BDNF、b FGF的影響,明確頭穴透刺預(yù)處理對(duì)腦缺血性卒中的預(yù)防與治療作用,從干細(xì)胞這一角度,探討頭穴透刺預(yù)處理預(yù)防缺血性腦卒中的可能機(jī)制。方法:實(shí)驗(yàn)選取雄性Wistar大鼠120只,采用隨機(jī)數(shù)字表方法將大鼠隨機(jī)分為:假手術(shù)組、模型組、實(shí)驗(yàn)組,每組各40只,每組又分為3、7、14、21d四個(gè)時(shí)間點(diǎn),每個(gè)時(shí)間點(diǎn)10只;采用改良線栓法制備大鼠腦缺血再灌注模型(MCAO),實(shí)驗(yàn)組于造模前連續(xù)針刺7d,針刺選取百會(huì)透雙側(cè)太陽穴,每次針刺治療時(shí)間為20分鐘,針刺治療后造模,模型組僅造模,不做其他處理,假手術(shù)組除不插線外,余操作同模型組;通過腹腔注射Brdu來標(biāo)記增殖的細(xì)胞,分別于3、7、14、21d四個(gè)時(shí)間點(diǎn)取材來對(duì)實(shí)驗(yàn)對(duì)象進(jìn)行觀察研究。采用神經(jīng)癥狀學(xué)評(píng)分確定模型成功標(biāo)準(zhǔn),通過TTC染色和HE染色來了解腦缺血再灌注后腦組織的病理學(xué)改變。通過免疫組化觀察Brdu、Nestin陽性細(xì)胞,確定NSCs的增殖情況,通過免疫熒光化學(xué)技術(shù)觀察Brd U/GFAP、Brd U/NSE雙標(biāo)細(xì)胞,確定NSCs分化情況。免疫組化法觀察BDNF、b FGF變化,確定對(duì)NSCs增殖的可能機(jī)制。結(jié)果:1.實(shí)驗(yàn)組與模型組各個(gè)時(shí)間點(diǎn)比較神經(jīng)功能缺損評(píng)分均降低,實(shí)驗(yàn)組在7、14、21d點(diǎn)神經(jīng)功能評(píng)分與模型組比較具有顯著性差異(P0.05)。2.實(shí)驗(yàn)組各個(gè)時(shí)間點(diǎn)與模型組比較Brd U免疫陽性細(xì)胞表達(dá)均增強(qiáng),并隨時(shí)間變化,3d時(shí)開始增加,7d時(shí)達(dá)到高峰,14d開始下降,各個(gè)時(shí)間點(diǎn)陽性細(xì)胞數(shù)量與模型組相比,有顯著性差異(P0.05)。3.實(shí)驗(yàn)組Nestin免疫陽性細(xì)胞表達(dá)明顯增強(qiáng),3d時(shí)開始增加,7d時(shí)達(dá)到高峰,7、14、21d與模型組陽性細(xì)胞數(shù)相比,有顯著性差異(P0.05)。4.實(shí)驗(yàn)組Brd U/GFAP、Brd U/NSE免疫熒光雙標(biāo)陽性細(xì)胞明顯增加,于缺血再灌注后7d達(dá)高峰,14d開始下降,與模型組7、14d比較有顯著性差異(P0.05)。5.實(shí)驗(yàn)組BDNF的表達(dá)明顯增強(qiáng),于缺血再灌注21d時(shí)達(dá)高峰,與模型組7、14、21d相比有顯著性差異(P0.05)。6.實(shí)驗(yàn)組b FGF的表達(dá)明顯增強(qiáng),于7d時(shí)達(dá)高峰。與模型組各個(gè)時(shí)間點(diǎn)比較有顯著性差異(P0.05)。結(jié)論:1、頭穴透刺預(yù)處理能顯著降低腦缺血再灌注后大鼠神經(jīng)功能癥狀評(píng)分,促進(jìn)其神經(jīng)功能恢復(fù)。2、頭穴透刺預(yù)處理能顯著促進(jìn)腦缺血再灌注后大鼠腦組織Brdu、Nestin、Brd U/GFAP、Brd U/NSE表達(dá)增加,從而促進(jìn)NSCs的增殖和分化。3、頭穴透刺預(yù)處理能促進(jìn)腦缺血再灌注后大鼠神經(jīng)營(yíng)養(yǎng)因子b FGF、BDNF的表達(dá),從而有助于神經(jīng)功能的恢復(fù)。
[Abstract]:Objective: to investigate the effects of scalp acupoint penetration preconditioning on the proliferation and differentiation of endogenous NSCs after cerebral ischemia-reperfusion in rats and the effects on BDNF bFGF according to the idea of "treating disease without disease" in traditional Chinese medicine (TCM). To clarify the preventive and therapeutic effects of scalp acupoint penetration preconditioning on ischemic stroke and to explore the possible mechanism of scalp acupoint penetration preconditioning in preventing ischemic stroke from the point of view of stem cells. Methods: 120 male Wistar rats were selected and randomly divided into four groups: sham operation group, model group, experimental group, 40 rats in each group. The rat model of cerebral ischemia-reperfusion (MCAO) was established by modified thread embolization method. The experimental group was treated continuously for 7 days before the model was made. The acupuncture group was treated with Baihui dialysed bilateral temples for 20 minutes each time. The model was made after acupuncture treatment, and only the model was made in the model group. Without any other treatment, the sham-operation group operated with the same model group except without wire insertion, and the proliferating cells were labeled by intraperitoneal injection of Brdu. The successful criteria of the model were determined by neurosymptomatic score, and the pathological changes of brain tissue after cerebral ischemia and reperfusion were studied by TTC staining and HE staining. The proliferation of NSCs was determined by immunohistochemical observation of Brdun nestin positive cells, and the differentiation of NSCs was determined by using immunofluorescence technique to observe BrdU / GFAP / Brd / NSE double labeled cells. The changes of BDNFB FGF were observed by immunohistochemical method to determine the possible mechanism of proliferation of NSCs. The result is 1: 1. Compared with the model group, the neurological deficit score of the experimental group and the model group decreased at each time point, and there was significant difference between the experimental group and the model group on the 21st day (P0.05). 2. Compared with the model group, the expression of BrdU immunoreactive cells in the experimental group increased at each time point, and increased at the 3rd day and reached the peak at the 7th day. The number of the positive cells in each time point was significantly different from that in the model group (P0.05). 3. The expression of BrdU immunoreactive cells in the experimental group was significantly higher than that in the model group (P0.05). The expression of nestin immunoreactive cells in the experimental group was significantly higher than that in the model group (P0.05). BrdU / GFAPG BrdU / NSE double immunofluorescence positive cells increased significantly in the experimental group, and began to decrease on the 7th day after ischemia and reperfusion, reaching the peak at the 14th day, which was significantly different from that in the model group (P0.05) on the 14th day (P0.05). The expression of BDNF in the experimental group was significantly increased and reached its peak at 21 days after ischemia reperfusion, which was significantly different from that in the model group on the 21st day (P0.05). The expression of bFGF in the experimental group was significantly increased and reached its peak at 7 days. There was significant difference between the model group and each time point (P0.05). Conclusion the scalp point penetration preconditioning can significantly decrease the neurological function symptom score and promote the recovery of nerve function after cerebral ischemia reperfusion in rats. The scalp penetration acupuncture preconditioning can significantly increase the expression of Brd Nestin + Brd / GFAPP + Brd / NSE in brain tissue after cerebral ischemia and reperfusion. In order to promote the proliferation and differentiation of NSCs, scalp acupoint penetration preconditioning can promote the expression of neurotrophic factor b FGFN BDNF after cerebral ischemia-reperfusion in rats, thus contributing to the recovery of nerve function.
【學(xué)位授予單位】：黑龍江省中醫(yī)藥科學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R245

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