本文選題：視神經(jīng)損傷 + 晶狀體損傷��； 參考：《鄭州大學(xué)》2012年碩士論文

【摘要】：視神經(jīng)的損傷(optic nerve injury)在眼科的臨床工作中很常見。視神經(jīng)損傷后往往預(yù)后很差,據(jù)統(tǒng)計(jì)致盲率可達(dá)40%～50%。視神經(jīng)損傷后的再生很難,因此,探討視神經(jīng)損傷后再生困難的機(jī)制以及尋找有效方法促進(jìn)視神經(jīng)再生是廣大學(xué)者研究的焦點(diǎn)。傳統(tǒng)觀點(diǎn)提出,成年哺乳動(dòng)物的中樞神經(jīng)損傷后沒有再生能力而只有再生反應(yīng),Aguage在實(shí)驗(yàn)中發(fā)現(xiàn),將坐骨神經(jīng)移植到視網(wǎng)膜,可明顯提高神經(jīng)節(jié)細(xì)胞(Retina ganglion cells, RGCs)的存活率,從而促進(jìn)損傷后的視神經(jīng)修復(fù)再生,這一實(shí)驗(yàn)證明視神經(jīng)損傷后可再生。 近年來的大量實(shí)驗(yàn)研究發(fā)現(xiàn),損傷視神經(jīng)的同時(shí)損傷晶狀體,并成功觀察到損傷后的晶狀體形成外傷性白內(nèi)障,可成功提高視網(wǎng)膜神經(jīng)節(jié)細(xì)胞的存活機(jī)率,從而促進(jìn)視神經(jīng)的修復(fù)再生。Fischer[2]與Leon兩個(gè)小組的研究工作表明其可能的機(jī)制與神經(jīng)營(yíng)養(yǎng)因子(NGF、 BDNF、CNTF等)、局部炎癥、晶狀體蛋白等有關(guān)。近年來,隨著髓鞘相關(guān)抑制分子Nogo-A[3]、 MAG[4][5]、 OMGP[6]等相繼被證實(shí),晶狀體損傷對(duì)視神經(jīng)損傷后再生的促進(jìn)作用的機(jī)制是否與這些神經(jīng)生長(zhǎng)抑制因子有關(guān)報(bào)道很少,本實(shí)驗(yàn)擬對(duì)此相關(guān)機(jī)制進(jìn)行探討。 目的 觀察單純視神經(jīng)損傷及視神經(jīng)損傷聯(lián)合晶狀體損傷后視神經(jīng)中Nogo-AmRNA及Nogo-A的表達(dá),探討晶狀體損傷促進(jìn)視神經(jīng)損傷后再生的機(jī)制 方-法 實(shí)驗(yàn)動(dòng)物選取Wistar大鼠,共66只。隨即分為三組：A組：正常對(duì)照組(6只);B組：?jiǎn)渭円暽窠?jīng)損傷組(30只)；C組：視神經(jīng)損傷聯(lián)合晶狀體損傷組(30只)。分別于7d、14d、21d處死大鼠2只(A組)、10只(B組)、10只(C組),光鏡下觀察視神經(jīng)的病理變化,免疫組化染色檢測(cè)Nogo-a表達(dá)情況,采用逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)(RT-PCR),半定量分析不同組視神經(jīng)Nogo-A mRNA的表達(dá) 結(jié)果 1.視神經(jīng)的少突膠質(zhì)細(xì)胞中Nogo-a的表達(dá) 損傷后7天,正常視神經(jīng)對(duì)照組中,Nogo-A染色陽性的少突膠質(zhì)細(xì)胞單行排列,排列規(guī)則,陽性細(xì)胞數(shù)為：117.40±2.84,呈淡藍(lán)色。術(shù)后第7天,單純損傷組神經(jīng)纖維排列不規(guī)則,少突膠質(zhì)細(xì)胞排列紊亂,陽性細(xì)胞數(shù)為：136.80±3.94,呈深藍(lán)色。視神經(jīng)聯(lián)合晶狀體損傷組神經(jīng)纖維排列不規(guī)則,少突膠質(zhì)細(xì)胞排列錯(cuò)亂,陽性細(xì)胞數(shù)：130.40±2.48,呈深藍(lán)色。3組中Nogo-A表達(dá)陽性細(xì)胞數(shù)進(jìn)行兩兩比較,除視神經(jīng)損傷聯(lián)合晶狀體損傷組與單純損傷組間差異無統(tǒng)計(jì)學(xué)意義(P=-0.277)外,其余每?jī)山M之間的比較都有統(tǒng)計(jì)學(xué)意義。 損傷后14天,陽性細(xì)胞計(jì)數(shù)在正常視神經(jīng)組為：119.80±4.45,單純視神經(jīng)損傷組為：128.40±3.82,視神經(jīng)損傷聯(lián)合晶狀體損傷組為：122.6±3.23。單純損傷組與正常喂養(yǎng)組比較差異有統(tǒng)計(jì)學(xué)意義(P0.05)、聯(lián)合損傷組與正常喂養(yǎng)組比較差異無統(tǒng)計(jì)學(xué)意義(P=0.632),單純損傷組與聯(lián)合損傷組之間差異無統(tǒng)計(jì)學(xué)意義(P=0.121)。 損傷后21天,正常組陽性細(xì)胞數(shù)為：116.60±4.38,單純視神經(jīng)損傷組陽性細(xì)胞數(shù)為：130.40±2.48,視神經(jīng)損傷聯(lián)合晶狀體損傷組為：104.40±6.78。三組之間每?jī)山M之間的比較差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。 2.樣本經(jīng)PCR擴(kuò)增、瓊脂糖凝膠電泳后顯示Nogo-A mRNA在單純損傷組和聯(lián)合損傷組均可檢測(cè)到,且在術(shù)后7d、14d和21d, LabWorks軟件分析顯示Nogo-A mRNA的擴(kuò)增表現(xiàn)出很穩(wěn)定的表達(dá)水平。 結(jié)論 1.正常視神經(jīng)也可見Nogo-a和Nogo-A mRNA的表達(dá),且三個(gè)時(shí)間點(diǎn)表達(dá)較穩(wěn)定。 2.單純視神經(jīng)損傷組及聯(lián)合損傷組后Nogo-A表的陽性細(xì)胞數(shù)明顯增多。 3.晶體狀損傷促進(jìn)軸突再生的機(jī)制與Nogo-A有關(guān),可能是晶狀體損傷后阻礙了Nogo-A mRNA的轉(zhuǎn)錄過程或者影響了Nogo-A的穩(wěn)定性。
[Abstract]:Optic nerve injury is very common in the clinical work of the ophthalmology. The prognosis is very poor after optic nerve injury. It is difficult to regenerate after 40% ~ 50%. optic nerve injury according to the statistical blinding rate. Therefore, it is a study to explore the mechanism of the difficulty of regeneration after optic nerve injury and to find an effective method to promote the optic nerve regeneration. The traditional point of view suggests that the central nerve injury of adult mammals has no regeneration ability but only regenerative response. In the experiment, Aguage found that the transplantation of the sciatic nerve to the retina could obviously improve the survival rate of the ganglion cells (Retina ganglion cells, RGCs), thus promoting the regeneration of the optic nerve after the injury. To verify the regeneration of the optic nerve injury.
In recent years, a large number of experimental studies have found that the damage of the optic nerve at the same time damage the lens, and the successful observation of the traumatic cataract after the injury, can successfully improve the survival probability of retinal ganglion cells, thus promoting the repair and regeneration of the optic nerve,.Fischer[2] and Leon, two groups of research work to show that the possible machine It is related to NGF (BDNF, CNTF, etc.), local inflammation, and lens protein. In recent years, with myelin related inhibitors, Nogo-A[3], MAG[4][5], and OMGP[6] have been confirmed, whether the mechanism of lens injury to promote the regeneration of optic nerve injury is not related to these nerve growth inhibitors. The experiment is to discuss the related mechanism.
objective
To observe the expression of Nogo-AmRNA and Nogo-A in optic nerve after optic nerve injury and optic nerve injury combined with lens injury, and to explore the mechanism of lens injury to promote the regeneration of optic nerve after injury.
Square method
The experimental animals selected a total of 66 Wistar rats, and then divided into three groups: group A: normal control group (6); group B: simple optic nerve injury group (30); group C: optic nerve injury combined with lens injury group (30). 2 rats were killed in 7d, 14d, 21d, 10 (B group), 10 (C group). The pathological changes of optic nerve were observed under the light microscope, immunologic changes were observed under the light microscope. Immunity was observed under the light microscope. Immunity was observed under the light microscope. Immunology was observed under the light microscope. Immunity was observed under the light microscope, immune to the pathological changes of the optic nerve under the light microscope to observe the immune pathological changes under the light scope, immunization under the light scope, immunity under the observation of the immune system The expression of Nogo-a was detected by histochemical staining, and the expression of Nogo-A mRNA in different groups was analyzed by reverse transcription polymerase chain reaction (RT-PCR).
Result
Expression of Nogo-a in the oligodendrocytes of 1. optic nerve
In the normal optic nerve control group 7 days after the injury, the Nogo-A positive oligodendrocytes were arranged in a single row and arranged regularly. The number of positive cells was 117.40 + 2.84, and the number of oligodendrocytes in simple injury group was irregular and the oligodendrocytes were arranged irregular, and the number of positive cells was 136.80 + 3.94, and the optic nerve was dark blue. The arrangement of nerve fibers in the combined lens injury group was irregular and the oligodendrocytes were arranged in disorder. The number of positive cells was 130.40 + 2.48. The number of Nogo-A positive cells in the group of deep blue.3 was compared, and the difference between the optic nerve injury combined with the lens injury group and the simple injury group was not statistically significant (P=-0.277), and the other two groups were in each group. There is statistical significance in the comparison.
14 days after injury, the number of positive cells in the normal optic nerve group was 119.80 + 4.45, and the only optic nerve injury group was 128.40 + 3.82. The optic nerve injury combined with the lens injury group was 122.6 + 3.23. simple injury group and the normal feeding group was statistically significant (P0.05). There was no statistical difference between the combined injury group and the normal feeding group. Significance (P=0.632), there was no significant difference between pure injury group and combined injury group (P=0.121).
21 days after injury, the number of positive cells in the normal group was 116.60 + 4.38, and the number of positive cells in the simple optic nerve injury group was 130.40 + 2.48, and the optic nerve injury combined with the lens injury group was statistically significant (P0.05) between the 104.40 + 6.78. three groups.
2. samples were amplified by PCR, and the agarose gel electrophoresis showed that Nogo-A mRNA could be detected in both simple injury group and joint injury group, and 7d, 14d and 21d after operation. LabWorks software analysis showed that the amplification of Nogo-A mRNA showed a stable expression level.
conclusion
1. the expression of Nogo-a and Nogo-A mRNA was also observed in normal optic nerve, and the expression was stable at three time points.
2. the number of Nogo-A positive cells in the optic nerve injury group and the combined injury group increased significantly.
The mechanism of 3. crystal injury promoting axon regeneration is related to Nogo-A, which may be hindering the transcriptional process of Nogo-A mRNA or affecting the stability of Nogo-A after lens injury.

【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R774.6

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