本文選題：腦源性神經(jīng)營養(yǎng)因子 + 視網(wǎng)膜神經(jīng)節(jié)細(xì)胞�。� 參考：《重慶醫(yī)科大學(xué)》2011年碩士論文

【摘要】：目的 觀察超聲微泡造影劑介導(dǎo)腦源性神經(jīng)營養(yǎng)因子(BDNF)聯(lián)合轉(zhuǎn)染大鼠視網(wǎng)膜和視皮質(zhì)區(qū)細(xì)胞對(duì)視神經(jīng)損傷后視網(wǎng)膜神經(jīng)節(jié)細(xì)胞(RGC)的保護(hù)作用。 方法 雄性Sprague-Dawley(SD)大鼠88只隨機(jī)分為正常組(A組)、假手術(shù)組(B組)、空白對(duì)照組(C組)、單純眼轉(zhuǎn)染組(D組)、單純腦轉(zhuǎn)染組(E組)、聯(lián)合轉(zhuǎn)染組(F組)。A組8只大鼠,B～F組每組16只大鼠。建立鉗夾視神經(jīng)損傷模型,將B～F組隨機(jī)分為視神經(jīng)損傷1、2周亞組,各亞組8只大鼠。B、C組分別玻璃體腔和視皮質(zhì)區(qū)注射磷酸鹽緩沖液(PBS),D、E組分別玻璃體腔和視皮質(zhì)區(qū)注射BDNF質(zhì)粒(pBDNF)微泡造影劑懸液,F組玻璃體腔和視皮質(zhì)區(qū)同時(shí)注射pBDNF微泡造影劑懸液。D～F組注射pBDNF微泡微泡造影劑懸液后,立即用超聲輻照相應(yīng)轉(zhuǎn)染部位。視神經(jīng)損傷1、2周,各組行逆行熒光金標(biāo)記RGC計(jì)數(shù);半胱氨酸蛋白酶-3(caspase-3)蛋白免疫組織化學(xué)染色,觀察其陽性表達(dá)情況;視網(wǎng)膜電流圖(PERG)檢測,記錄N95振幅。 結(jié)果 熒光金標(biāo)記檢測顯示F組視神經(jīng)損傷1、2周亞組RGC數(shù)分別為409.1±42.10個(gè)、390.8±35.34個(gè),占A組的86.0%、82.1%,均明顯高于C、D、E組,差異具有統(tǒng)計(jì)學(xué)意義(P㩳0.01)。F組各時(shí)間點(diǎn)Caspase-3陽性細(xì)胞數(shù)均少于C、D、E組,且染色較淺。PERG檢測中視神經(jīng)損傷1周、2周時(shí)F組N95振幅分別為7.57±0.66μV、7.33±0.58μV,高于C、D、E組(P㩳0.01),且與A組無明顯差異(P0.05)。 結(jié)論 超聲微泡造影劑介導(dǎo)BDNF基因聯(lián)合轉(zhuǎn)染視網(wǎng)膜和視皮質(zhì)區(qū)細(xì)胞能抑制視神經(jīng)損傷后RGC凋亡,提高RGC存活數(shù),還可能具有一定的電生理功能保護(hù)作用。
[Abstract]:Purpose To observe the protective effect of combined transfection of brain derived neurotrophic factor (BDNF) with ultrasound microbubble contrast medium on retinal ganglion cells (RGCs) after optic nerve injury in rats. Method 88 male Sprague-Dawley SD rats were randomly divided into normal group (n = 8), sham operation group (n = 8), control group (n = 16), control group (n = 8), group D (n = 8), group E (n = 8), group A (n = 8), group B (n = 8). The model of optic nerve injury by clamp was established, and the group B F was randomly divided into the group of 1 ~ 2 weeks after optic nerve injury. Eight rats in each subgroup were injected with BDNF plasmid pBDNF in vitreous body cavity and visual cortex, respectively. PBDNF was injected into vitreous body cavity and visual cortex respectively. PBDNF was injected into visual cortex and vitreous cavity of F group. After injection of pBDNF microbubble contrast agent suspension, the microbubble contrast agent suspension. The corresponding transfection site was immediately irradiated by ultrasound. After 1 and 2 weeks of optic nerve injury, the number of RGC labeled with retrograde fluorescence gold, the immunohistochemical staining of cysteine protease -3caspase-3) and the positive expression of cysteine protease -3caspase-3 were observed in each group, and the amplitude of N95 was recorded by electroretinogram (ERG). Result The number of RGC in group F was 409.1 鹵42.10 (390.8 鹵35.34) at 1 and 2 weeks after optic nerve injury, which accounted for 82.1% of group A, which was significantly higher than that in group E (P < 0.01). The number of Caspase-3 positive cells in group F was lower than that in group E at each time point. The N95 amplitudes of group F were 7.57 鹵0.66 渭 V, 7.33 鹵0.58 渭 V, respectively, which were higher than those of group A (P 0.01), and there was no significant difference between group A and group A. Conclusion The combined transfection of BDNF gene into retinal and visual cortex cells mediated by ultrasound microbubble contrast agent can inhibit the apoptosis of RGC after optic nerve injury and increase the survival of RGC. It may also have a protective effect on electrophysiological function.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R774.1

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 陳玲玲;超聲波結(jié)合微泡技術(shù)聯(lián)合間充質(zhì)干細(xì)胞重建做功心肌的實(shí)驗(yàn)研究[D];川北醫(yī)學(xué)院;2013年

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本文編號(hào)：1841964