本文選題：針刺 + 單眼視覺剝奪　； 參考：《長春中醫(yī)藥大學(xué)》2014年碩士論文

【摘要】：目的：通過運(yùn)用免疫組化方法、激光共聚焦技術(shù)觀察針刺對(duì)可塑期內(nèi)單眼視覺剝奪大鼠視網(wǎng)膜神經(jīng)元中相關(guān)生長蛋白（GAP-43）結(jié)構(gòu)發(fā)育的干預(yù)效應(yīng)，從而進(jìn)一步討論弱視發(fā)病機(jī)制與針刺對(duì)視覺系統(tǒng)可塑期內(nèi)的調(diào)節(jié)作用。為針刺治療弱視疾病的機(jī)制提供新的思路，為臨床治療弱視的提供參考方法。 方法：使用出生后14天清潔級(jí)實(shí)驗(yàn)大鼠60只，采用隨機(jī)分組的方法將實(shí)驗(yàn)大鼠分為五組，即正常組、模型組、早期針刺組、中期針刺組、晚期針刺組，將模型組和早期針刺組、中期針刺組、晚期針刺組采用縫合單側(cè)上下眼瞼的方法復(fù)制單眼剝奪弱視模型；正常組不給予任何治療處理，五組實(shí)驗(yàn)大鼠全都放在一樣的環(huán)境中飼養(yǎng)。 早期針刺組在單眼剝奪大鼠模型復(fù)制后的第7天開始進(jìn)行針刺治療；中期針刺組在單眼剝奪大鼠模型復(fù)制后的第14天開始進(jìn)行針刺治療；晚期針刺組在單眼剝奪大鼠模型復(fù)制后的第21天開始進(jìn)行針刺治療。選取“睛明”、“攢竹”、“光明”、“風(fēng)池”四個(gè)穴位，在針刺治療結(jié)束后進(jìn)行P-VEP檢測(cè)，觀察檢測(cè)視覺誘發(fā)電位（P-VEP）和利用免疫組化方法檢測(cè)目標(biāo)蛋白在視網(wǎng)膜中成像；剝奪早期針刺組、剝奪中期針刺組在針刺結(jié)束后進(jìn)行檢測(cè)。 麻醉實(shí)驗(yàn)動(dòng)物取弱視大鼠剝奪眼視網(wǎng)膜，用4%多聚甲醛浸泡視網(wǎng)膜10-20分鐘，然后每隔1-2小時(shí)分別換10%、20%、30%蔗糖，30%蔗糖后置于4攝氏度過夜將在4攝氏度過夜后的大鼠視網(wǎng)膜進(jìn)行包埋冷凍切片視網(wǎng)膜，視網(wǎng)膜取厚度14微米切片，貼于硅化防脫載玻片上，，免疫組織化學(xué)方法觀察GAP-43在組織中的陽性表達(dá)，最后使用激光掃描共聚焦顯微鏡觀察目標(biāo)蛋白（GAP-43）細(xì)胞熒光強(qiáng)度變化。 結(jié)果： (1)在進(jìn)行單眼視覺剝奪模型復(fù)制4周后，模型組P-VEP波形有明顯改變，表現(xiàn)為P100的明顯延遲和N45-P100幅值降低，其與正常組的波形比較有較為明顯的差異（P＜0.01）；針刺治療后，單眼視覺剝奪針刺組與模型組相比時(shí)值與幅值均有顯著變化，其P-VEP波形P100出現(xiàn)的時(shí)值明顯提前（P＜0.05），N45-P100幅值明顯升高（P＜0.05） (2)視網(wǎng)膜：正常組和模型組相比，大鼠視網(wǎng)膜GAP-43陽性神經(jīng)元的表達(dá)顯著下降，有顯著性差異（P＜0.01）；針刺組與模型組比較，大鼠視網(wǎng)膜GAP-43陽性神經(jīng)元表達(dá)水平均明顯提高，有顯著性差異（P＜0.05）。 結(jié)論： （1）實(shí)驗(yàn)發(fā)現(xiàn)針刺能改善單眼視覺剝奪大鼠異常的P-VEP波形；單眼剝奪大鼠的P-VEP波形變化和視覺系統(tǒng)的相關(guān)生長蛋白GAP-43的表達(dá)降低有關(guān)。 （2）針刺對(duì)視覺發(fā)育敏感期內(nèi)單眼剝奪組大鼠視網(wǎng)膜GAP-43陽性細(xì)胞表達(dá)有調(diào)節(jié)作用，對(duì)視功能的恢復(fù)具治療的意義。 （3）針刺在視覺發(fā)育關(guān)鍵期內(nèi)干預(yù)單眼剝奪大鼠其視網(wǎng)膜中GAP-43陽性表達(dá)升高，提示針刺對(duì)弱視視功能有改善機(jī)制,GAP-43與視覺發(fā)育及可塑性有密切關(guān)系。
[Abstract]:Objective: to observe the effect of acupuncture on the development of GAP-43 in retinal neurons of rats with single eye visual deprivation during the plastic period by using immunohistochemical method and laser confocal technique. The mechanism of amblyopia and the regulating effect of acupuncture on the plastic period of visual system were discussed. It provides a new idea for the treatment of amblyopia and a reference method for clinical treatment of amblyopia. Methods: sixty clean grade experimental rats were randomly divided into five groups: normal group, model group, early acupuncture group, metaphase acupuncture group, late acupuncture group, model group and early acupuncture group. The amblyopia model of unilateral upper and lower eyelids was made by suturing the upper and lower eyelids in the middle period acupuncture group and the late stage acupuncture group, while the normal group was not treated with any treatment, and the five experimental rats were all fed in the same environment. In the early acupuncture group, acupuncture treatment began on the 7th day after the establishment of the monocular deprivation rat model, and the acupuncture treatment began on the 14th day after the establishment of the monocular deprivation model in the metaphase acupuncture group. The late acupuncture group was treated with acupuncture on the 21 th day after the rat model of monocular deprivation was made. Four acupoints of "Jingming", "Zanzhu", "Guangming" and "Fengchi" were selected. After acupuncture treatment, P-VEP was detected, visual evoked potential (VEP) was detected and target protein was detected in retina by immunohistochemical method. The early acupuncture group was deprived and the mid-stage acupuncture group was tested after acupuncture. Rats with amblyopia were taken from anesthetized rats and deprived of their retina. The retina was soaked in 4% paraformaldehyde for 10-20 minutes. Then every 1-2 hours, 10% 30% sucrose 30% sucrose and 30% sucrose were replaced and placed overnight at 4 degrees Celsius. The retina of the rats after the night was embedded in frozen sections, and the retina was cut 14 microns thick, and applied to the silicified anti-desorption slide glass. Immunohistochemical method was used to observe the positive expression of GAP-43 in tissues. The fluorescence intensity of target protein (GAP-43) cells was observed by laser scanning confocal microscopy (LSCM). Results: (1) after 4 weeks of monocular visual deprivation model replication, the P-VEP waveform of the model group was significantly changed, showing that the P100 delayed and the N45-P100 amplitude decreased, which was significantly different from that of the normal group (P < 0.01), and after acupuncture treatment, there was a significant difference between the two groups (P < 0.01), and there was no significant difference between the two groups (P < 0.01). Compared with the model group, the time value and amplitude of the P-VEP wave P100 in the monocular visual deprivation acupuncture group were significantly changed, and the amplitude of the P-VEP waveform P100 was significantly higher than that of the model group (P < 0.05) (P < 0.05). The amplitude of N45-P100 in the monocular visual deprivation group was significantly higher than that in the model group (P < 0.05). (2) retina: compared with the model group, the expression of GAP-43 positive neurons in the normal group was significantly lower than that in the model group (P < 0.01), and the expression level of GAP-43 positive neurons in the retina of the acupuncture group was significantly higher than that in the model group. There was significant difference (P < 0.05). Conclusion: The results showed that acupuncture could improve the abnormal P-VEP waveforms of monocular visual deprivation rats, and the changes of P-VEP waveforms in monocular deprived rats were related to the decreased expression of growth protein GAP-43 in the visual system. (2) Acupuncture can regulate the expression of GAP-43 positive cells in the retina of the monocular deprivation group during the sensitive period of visual development, and has the therapeutic significance for the recovery of visual function. 3) the positive expression of GAP-43 in the retina of monocular deprived rats was increased after acupuncture intervention during the critical period of visual development, suggesting that acupuncture can improve the function of amblyopia. GAP-43 is closely related to visual development and plasticity.
【學(xué)位授予單位】：長春中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R245

【參考文獻(xiàn)】

相關(guān)期刊論文 前6條

1 嚴(yán)恒林;沈馨亞;;生長相關(guān)蛋白(GAP-43)與神經(jīng)發(fā)育和再生[J];神經(jīng)解剖學(xué)雜志;1993年02期

2 邵毅;繆超英;余瑤;裴重剛;周瓊;譚鋼;高桂平;彭娟;董雯佳;;艾炙風(fēng)池穴聯(lián)合遮蓋療法治療兒童單眼弱視[J];眼科新進(jìn)展;2012年01期

3 朱曉明,嚴(yán)宏;視覺發(fā)育敏感期的研究進(jìn)展[J];眼視光學(xué)雜志;2004年04期

4 孔德蘭,李超,杜學(xué)齡,王曉麗,楊華;屈光不正兒童立體視覺敏感期臨床研究[J];中國實(shí)用眼科雜志;2003年03期

5 張毅;李海波;汪澤;;兒童弱視治療前后VEP、ERG檢查的臨床意義的探討[J];中國實(shí)用眼科雜志;2006年12期

6 周英;;攢竹穴針刺方法及臨床應(yīng)用[J];針灸臨床雜志;2010年07期

本文編號(hào)：1804209