本文關(guān)鍵詞：針刺和神經(jīng)干細(xì)胞聯(lián)合治療實驗性腦性癱瘓的機制研究　出處：《廣州醫(yī)學(xué)院》2011年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 針刺 神經(jīng)干細(xì)胞 腦性癱瘓

【摘要】：目的: 探討針刺和神經(jīng)干細(xì)胞(Neural stem cell, NSC)聯(lián)合治療對缺血缺氧性腦損傷(Hypoxic-ischemic brain damage,HIBD)幼鼠肢體運動功能、學(xué)習(xí)和記憶能力、腦組織凋亡基因bax及抗凋亡基因bcl-2的蛋白表達和病理形態(tài)學(xué)的影響,并對針刺聯(lián)合NSC療法治療實驗性腦性癱瘓(Cerebral palsy,CP)的機制進行探索,為臨床小兒CP的治療提供實驗依據(jù)。 內(nèi)容和方法: 1、實驗動物及分組:7日齡SPF級SD幼鼠68只(體重:12g～20g),隨機選取12只作為①正常對照組;HIBD成功模型隨機分成以下4組:②模型組;③單純針刺組;④單純NSC組;⑤針刺+NSC組,每組12只。 2、治療階段:包括針刺治療和NSC治療,其中NSC采用幼鼠尾靜脈注射。 3、行為學(xué)檢測:包括懸吊試驗、斜坡試驗和Y迷宮試驗。 4、取材:術(shù)后21天對正常組和實驗組幼鼠斷頸取材,用HE染色法觀察其海馬組織病理形態(tài)學(xué)改變,并計數(shù)海馬組織正常神經(jīng)細(xì)胞數(shù)目;用TUNEL法檢測其腦皮質(zhì)神經(jīng)細(xì)胞凋亡情況并測出凋亡神經(jīng)細(xì)胞數(shù)目;免疫組化法檢測其腦組織bcl-2、bax基因的蛋白質(zhì)表達。 5、數(shù)據(jù)整理,統(tǒng)計學(xué)分析。 結(jié)果: 1、行為學(xué)檢測:懸吊試驗、斜坡試驗、Y迷宮試驗中,③單純針刺組和④單純NSC組與②模型組相比,各項均數(shù)差異均無統(tǒng)計學(xué)意義(p 0.05),表明單純針刺和單純NSC均不能明顯改善幼鼠的肢體運動功能和學(xué)習(xí)記憶能力;⑤針刺+NSC組與②模型組相比,各項均數(shù)差異有統(tǒng)計學(xué)意義(p 0.05),表明針刺聯(lián)合NSC能顯著提高幼鼠的肢體運動功能和學(xué)習(xí)記憶能力(智力水平)。 2、bcl-2基因和bax基因的蛋白質(zhì)表達:⑤針刺和NSC組與②模型組相比,均數(shù)差異均有統(tǒng)計學(xué)意義(p 0.05),表明針刺聯(lián)合NSC能夠明顯促進抗凋亡基因bcl-2的蛋白質(zhì)表達、抑制凋亡基因bax的蛋白質(zhì)表達;③單純針刺組及④單純NSC組與②模型組相比差異均無統(tǒng)計學(xué)意義(p 0.05)。 3、腦組織肉眼觀察:取材可見②模型組左側(cè)腦組織出現(xiàn)大面積壞死,空泡,右側(cè)腦組織組織結(jié)構(gòu)正常;③單純針刺組、④單純NSC組、⑤針刺+NSC組左側(cè)腦組織都出現(xiàn)了不同程度的壞死和空泡區(qū)域,但程度都比②模型組較輕,其中⑤針刺+NSC組比③單純針刺組和④單純NSC組左側(cè)腦組織損傷程度更輕。 4、腦組織HE切片:與②模型組相比,③單純針刺組、④單純NSC組、⑤針刺+NSC組幼鼠海馬變性神經(jīng)細(xì)胞數(shù)量減少,病理形態(tài)學(xué)明顯改善,其中⑤針刺+NSC組海馬變性神經(jīng)細(xì)胞數(shù)目最少,腦組織病變程度最輕。 5、神經(jīng)細(xì)胞計數(shù):(1)海馬正常神經(jīng)細(xì)胞數(shù)目:②模型組與①正常組相比,均數(shù)更少,差異有統(tǒng)計學(xué)意義(p 0.05),表明HIBD后幼鼠海馬正常神經(jīng)細(xì)胞數(shù)目顯著減少;③單純針刺組和④單純NSC組與②模型組相比,均數(shù)差異均無統(tǒng)計學(xué)意義(p 0.05),表明單純針刺治療和單純NSC治療都不能明顯提高HIBD后幼鼠海馬正常神經(jīng)細(xì)胞數(shù)目;⑤針刺+NSC組與模型組相比,均數(shù)更多,差異有統(tǒng)計學(xué)意義(p 0.05),表明⑤針刺+NSC組可明顯提高HIBD后幼鼠海馬正常神經(jīng)細(xì)胞數(shù)目。(2)腦皮質(zhì)組織細(xì)胞凋亡:③單純針刺組和④單純NSC組與③模型組相比,均數(shù)差異均無統(tǒng)計學(xué)意義(p 0.05),表明單純針刺和單純NSC都不能明顯抑制幼鼠腦皮質(zhì)神經(jīng)細(xì)胞凋亡;⑤針刺+NSC組與②模型組相比,均數(shù)差異有統(tǒng)計學(xué)意義(p 0.05),表明針刺聯(lián)合NSC治療可明顯抑制幼鼠腦皮質(zhì)神經(jīng)細(xì)胞的凋亡。 結(jié)論: 1、單純針刺或單純NSC都不能明顯改善缺血缺氧性腦癱模型幼鼠肌力、隨意運動、學(xué)習(xí)記憶能力(智力)。 2、針刺和NSC聯(lián)合治療對缺血缺氧性腦癱模型幼鼠的肌力增長、隨意運動的恢復(fù)、學(xué)習(xí)和記憶能力(智力)的改善有明顯效果。 3、針刺和NSC聯(lián)合治療對缺血缺氧性腦癱模型幼鼠腦組織細(xì)胞凋亡基因bax的蛋白質(zhì)表達有明顯的抑制作用、對抗凋亡基因bcl-2的蛋白質(zhì)表達有明顯的促進作用。
[Abstract]:Objective:
To investigate the effects of acupuncture on neural stem cells (Neural and stem cell, NSC) injury combined treatment on hypoxic ischemic brain (Hypoxic-ischemic brain damage, HIBD) in limb motor function, learning and memory ability and brain tissue apoptosis gene Bax and anti apoptosis gene Bcl-2 protein expression and morphological changes, and paralysis of the acupuncture combined with NSC therapy treatment of experimental cerebral (Cerebral palsy CP) to explore the mechanism, to provide experimental basis for clinical treatment of children with CP.
Content and methods:
1, experimental animals and groups: 68 SPF rats (7 to 20g) at the age of 7 days (body weight: 12g to 20g), 12 were selected as normal control group, HIBD successful model was randomly divided into the following 4 groups: Group II, model group, 3 simple acupuncture group, 4 simple NSC group, and 12 +NSC group.
2, treatment stage, including acupuncture treatment and NSC treatment, of which NSC was injected into the tail vein of young rats.
3, behavioral test: including suspension test, slope test and Y labyrinth test.
4, material: 21 days after the operation, the normal group and experimental group of rats by cervical specimens, observe pathological changes in hippocampus by HE staining, and count the number of nerve cells in hippocampus of normal cerebral cortex; to detect the apoptosis of neural cells and measured the number of apoptotic nerve cells by TUNEL method; immunohistochemistry the brain tissue Bcl-2 protein expression of Bax gene.
5, data sorting, statistical analysis.
Result:
1, the behavior detection: suspension test, slope test, Y maze test, the acupuncture group compared with the NSC group and the model group, the mean differences were not statistically significant (P, 0.05) showed that acupuncture and NSC alone could not significantly improve rat limb movement function and learning and memory compared with the acupuncture ability; +NSC group and model group, the mean difference was statistically significant (P 0.05), indicating that acupuncture combined with NSC can significantly improve the rat limb motor function and the ability of learning and memory (Intelligence).
2, the protein expression of bcl-2 gene and Bax gene: the acupuncture and NSC group and the model group, the mean difference was statistically significant (P, 0.05) showed that the protein expression of acupuncture combined with NSC can significantly promote the anti apoptosis gene Bcl-2 and apoptosis inhibiting gene Bax protein expression; the acupuncture group and the simple the NSC group and the model group, the differences were not statistically significant (P 0.05).
3, to observe the brain tissue in model group were visible to naked eye: the left brain tissue showed a large area of necrosis, cavitation, right brain tissue structure was normal; the acupuncture group, the simple NSC group, the acupuncture group +NSC left brain tissue have suffered different degrees of necrosis and cavitation region, but the degree than in model group the lighter, which the +NSC acupuncture group than in the acupuncture group and the NSC group on the left side of brain injury degree lighter.
4, brain tissue HE staining: compared with the model group, the acupuncture group, the simple NSC group, the number of the acupuncture group +NSC hippocampus nerve cell degeneration reduced, pathological morphology significantly improved, in which the number of the acupuncture group +NSC hippocampal nerve cell degeneration at least, brain lesions lightest.
5, nerve cell count: (1) number of normal nerve cells in hippocampus: compared with the normal group, model group, the number was less, the difference was statistically significant (P, 0.05) showed that the number of normal nerve cells in hippocampus in rats after HIBD decreased significantly; the acupuncture group compared with the simple NSC group and model group. The mean differences were not statistically significant (P, 0.05) showed that acupuncture treatment and NSC treatment alone can significantly improve HIBD after normal nerve cells in hippocampus in rats of the acupuncture group +NSC number; compared with the model group, the number was more, the difference was statistically significant (P 0.05), the +NSC group showed that acupuncture can significantly improve HIBD after the normal nerve cells in hippocampus in rats (2). The number of apoptotic cells in the cerebral cortex tissue: the simple acupuncture group compared with the NSC group and the model group, the mean differences were not statistically significant (P, 0.05) showed that the simple acupuncture and simple NSC can inhibit Neuronal apoptosis in the cerebral cortex of young rats. 5. There was a significant difference between the acupuncture group +NSC and the model group (P 0.05), indicating that acupuncture combined with NSC treatment could significantly inhibit the apoptosis of neurons in the cerebral cortex of young rats.
Conclusion:
1, pure acupuncture or simple NSC can not significantly improve the muscle strength, random movement and learning and memory ability (Intelligence) of hypoxic cerebral palsy model young rats.
2, acupuncture combined with NSC has a significant effect on the increase of muscle strength, the recovery of voluntary movement, and the improvement of learning and memory ability in rats with hypoxic ischemic cerebral palsy.
3, acupuncture combined with NSC has a significant inhibitory effect on the protein expression of apoptosis gene Bax in the hypoxic cerebral palsy model rats, and significantly promotes the protein expression of the anti apoptotic gene bcl-2.

【學(xué)位授予單位】：廣州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R246.6;R742.3

【參考文獻】

相關(guān)期刊論文 前10條

1 祁巖超;劉振寰;;小兒腦癱的治療現(xiàn)狀與展望[J];中國實用神經(jīng)疾病雜志;2006年01期

2 姚靜嬋;胡國華;曹樟全;王凱旋;;高壓氧治療嬰幼兒腦癱的臨床療效觀察[J];浙江臨床醫(yī)學(xué);2007年10期

3 關(guān)麗君,楊虹;針刺治療腦癱患兒手功能障礙110例[J];遼寧中醫(yī)雜志;2005年03期

4 劉鄉(xiāng);柴鐵劬;孫娟;張瑞;孟令杰;趙丹;周健洪;陳東風(fēng);;靳三針療法對窒息腦癱幼鼠神經(jīng)干細(xì)胞增殖與分化的實驗研究[J];廣州中醫(yī)藥大學(xué)學(xué)報;2010年03期

5 施炳培;卜懷娣;李惠;史惟;楊紅;王素娟;侍孝娟;;針刺治療小兒腦性癱瘓精細(xì)運動功能障礙61例[J];上海針灸雜志;2007年09期

6 唐洪麗;;腦癱三個月內(nèi)治療最佳[J];醫(yī)藥與保健;2010年02期

7 龐源廣;袁瓊蘭;;神經(jīng)干細(xì)胞移植在缺血性腦損傷治療中的應(yīng)用[J];同濟大學(xué)學(xué)報(醫(yī)學(xué)版);2010年02期

8 高峰;楊小峰;鄭學(xué)勝;沈罡;劉偉國;;小鼠神經(jīng)干細(xì)胞免疫原性的研究[J];細(xì)胞與分子免疫學(xué)雜志;2010年04期

9 蘭新建;萬鳳鳳;;神經(jīng)干細(xì)胞移植在腦出血性疾病中的應(yīng)用[J];中國傷殘醫(yī)學(xué);2007年01期

10 梁松,吳洋,高春霞,陳沁璐,郭敏;頭針治療小兒腦性癱瘓104例療效分析[J];現(xiàn)代中西醫(yī)結(jié)合雜志;2004年01期

，

本文編號：1419959