Colivelin對(duì)視神經(jīng)保護(hù)作用的初步研究
本文選題:Colivelin + 外傷性視神經(jīng)病變; 參考:《眼科新進(jìn)展》2012年04期
【摘要】:目的初步探討Colivelin對(duì)大鼠外傷性視神經(jīng)損傷后的神經(jīng)保護(hù)作用和機(jī)理,及其保護(hù)效果是否具有劑量依賴性。方法 40只健康2月齡Wistar大鼠,隨機(jī)分為模型組、安慰劑組及Colivelin低劑量組(10-6 μmol·L-1 Colivelin)、Colivelin中劑量組(10-4μmol·L-1Colivelin)、Colivelin高劑量組(10-2 μmol·L-1 Colivelin),每組8只。40只大鼠均做單側(cè)視神經(jīng)損傷模型,模型組另一側(cè)未做任何處理的8眼為空白對(duì)照組。應(yīng)用無創(chuàng)血管夾建立大鼠視神經(jīng)夾傷模型,造模后30 min Colivelin治療組玻璃體內(nèi)分別注射不同濃度Colivelin 5 μL,安慰劑組注射5μLPBS緩沖液,模型組和空白對(duì)照組不給予任何治療。注藥后7d,通過視網(wǎng)膜切片技術(shù),進(jìn)行HE染色觀察視網(wǎng)膜神經(jīng)節(jié)細(xì)胞(retinal ganglion cell,RGC)的形態(tài)及數(shù)目,TUNEL染色法檢測(cè)RGC的凋亡以及免疫組織化學(xué)法檢測(cè)視網(wǎng)膜中caspase-3的表達(dá)。結(jié)果 HE染色可見空白對(duì)照組視網(wǎng)膜各層細(xì)胞排列整齊密集,模型組、安慰劑組、Colivelin治療組均可見部分RGC核固縮,但隨著Colivelin注射濃度的增加,RGC發(fā)生核固縮的數(shù)量減少。RGC計(jì)數(shù):空白對(duì)照組每400倍光鏡視野下RGC數(shù)量為25.750±1.264,模型組為8.236±1.239,安慰劑組為8.514±1.222,Colivelin低劑量組為14.500±1.021,Colivelin中劑量組為16.250±1.319,Colivelin高劑量組為18.097±1.323,除模型組與安慰劑組之間差異無統(tǒng)計(jì)學(xué)意義(P0.05)外,其他各組之間差異均有統(tǒng)計(jì)學(xué)意義(均為P0.05)。TUNEL染色:模型組和安慰劑組注藥后7 d TUNEL染色可見大量凋亡細(xì)胞,Colivelin治療組凋亡細(xì)胞數(shù)量隨Colivelin注射濃度的增加依次遞減。RGC細(xì)胞凋亡率:模型組為(60.928±2.961)%,安慰劑組為(61.446±2.755)%,Colivelin低劑量組為(58.432±2.835)%,Colivelin中劑量組為(51.948±2.802)%,Colivelin高劑量組為(47.656±2.331)%。Caspase-3表達(dá):模型組、安慰劑組及Colivelin低劑量組、中劑量組、高劑量組均可見caspase-3表達(dá)。平均光密度值顯示,模型組和安慰劑組相比caspase-3表達(dá)無明顯差異(分別為0.482±0.012和0.486±0.012),均高于Colivelin治療組(均為P0.05),并且Colivelin低劑量組、中劑量組、高劑量組組間差異亦均有統(tǒng)計(jì)學(xué)意義(分別為0.411±0.017、0.326±0.018、0.234±0.016;均為P0.05)。結(jié)論 Colivelin能有效增加視神經(jīng)損傷后RGC的數(shù)量,抑制其凋亡,減少caspase-3表達(dá),且這一作用呈劑量依賴性。
[Abstract]:Objective to investigate the neuroprotective effect and mechanism of Colivelin on traumatic optic nerve injury in rats and whether the protective effect is dose-dependent. Methods Forty healthy 2-month-old Wistar rats were randomly divided into three groups: model group, placebo group and Colivelin low dose group (10-6 渭 mol L-1 Colivelin). The model of unilateral optic nerve injury was established in 10 ~ 4 渭 mol L ~ (-1) Colivelin high-dose group (10 ~ (-2) 渭 mol ~ (-1) Colivelin). 8 eyes in the model group without any treatment on the other side were blank control group. A rat model of optic nerve injury was established by using a non-invasive vascular clamp. 30 min after the establishment of the model, different concentrations of Colivelin 5 渭 L were injected into the vitreous of the model group and 5 渭 L PBS buffer solution was injected into the placebo group. The model group and the blank control group were not given any treatment. At 7 days after injection, the morphology and number of retinal ganglion cells (retinal ganglion cells) were observed by HE staining. The apoptosis of retinal ganglion cells was detected by Tunel staining and the expression of caspase-3 in retina was detected by immunohistochemistry. Results HE staining showed that all layers of retinal cells in blank control group were arranged neatly and densely, and partial RGC nuclear contractions were observed in both the model group and the placebo group. However, with the increase of Colivelin injection concentration, the number of nuclear contractions in RGCs decreased. RGC count: the number of RGCs in the blank control group was 25.750 鹵1.264 per 400-fold light microscope, in the model group was 8.236 鹵1.239, and in the placebo group was 8.514 鹵1.222um Colivelin, 14.500 鹵1.021Colivelin in the middle dose group and 16.250 鹵1.319m Colivelin high dose group. There was no significant difference between the model group and the placebo group (P0.05). There were significant differences between the other groups (P0.05). Tunel staining: Tunel staining showed that the number of apoptotic cells decreased with the increase of Colivelin injection concentration in model group and placebo group 7 days after injection. RGC showed that the number of apoptotic cells in Colivelin treatment group decreased with the increase of Colivelin injection concentration. The rate of apoptosis in the model group was (60.928 鹵2.961) and in the placebo group was (61.446 鹵2.755). The expression of Caspase-3 in the low dose group was (58.432 鹵2.835) vs (51.948 鹵2.802) in the middle dose group and (47.656 鹵2.331) in the high dose group. Caspase-3 expression was observed in placebo group, Colivelin low dose group, middle dose group and high dose group. The mean optical density showed that there was no significant difference in caspase-3 expression between model group and placebo group (0.482 鹵0.012 and 0.486 鹵0.012, respectively), which was higher than that of Colivelin treatment group (P0.05). The difference of high dose group was also statistically significant (0.411 鹵0.017) 0.326 鹵0.018 ~ 0.234 鹵0.016; all were P0.05). Conclusion Colivelin can effectively increase the number of RGCs after optic nerve injury, inhibit their apoptosis and reduce the expression of caspase-3 in a dose-dependent manner.
【作者單位】: 廣西醫(yī)科大學(xué)第一附屬醫(yī)院眼科;
【分類號(hào)】:R774.6
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