本文選題：褪黑素 + 白內(nèi)障��； 參考：《山東大學(xué)》2010年碩士論文

【摘要】： 研究目的： 白內(nèi)障是一致盲眼病,氧化損傷是誘發(fā)白內(nèi)障的重要因素。褪黑素(MLT)是松果體分泌的一種吲哚類激素,是人體內(nèi)強(qiáng)有效的自由基清除劑及抗氧化劑。本課題應(yīng)用褪黑素作用于離體氧化損傷的大鼠晶狀體,通過體外抗氧化模型實(shí)驗(yàn),檢測(cè)其抗氧化能力,探討褪黑素對(duì)過氧化氫誘導(dǎo)大鼠白內(nèi)障形成的抑制作用,為尋求較為理想的抗氧化藥物提供理論依據(jù)。 研究方法： 體外取出發(fā)育正常的Wistar大鼠晶狀體,按隨機(jī)原則分為以下三組：A組為空白對(duì)照組,B組為過氧化氫(H2O2)處理組,C組為在B組的基礎(chǔ)上加入不同濃度的MLT,使MLT終濃度分別為10(C1組)、30(C2組)、50(C3組)、100(C4組)、200(C5組)μmol／L。體外培養(yǎng)24h后,觀察各組晶狀體的混濁程度,采用Image-ProPlus(IPP)6.0軟件分析晶狀體下黑色條帶的平均光密度值；酶標(biāo)儀測(cè)定各組晶狀體組織的蛋白含量、還原型谷胱甘肽(GSH)、丙二醛(MDA)、總抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、過氧化氫酶(CAT)含量。行HE染色倒置顯微鏡下觀察各組晶狀體上皮細(xì)胞的形態(tài)。 研究結(jié)果： 1.大鼠晶狀體混濁程度的觀察及定量分析培養(yǎng)結(jié)束后,裂隙燈下可見,A組晶狀體保持透明,晶狀體后灰線清晰可見；B組晶狀體全混呈白色,晶狀體后灰線模糊不清；C組晶狀體的透明性均高于B組,晶狀體后灰線呈不同程度的模糊。平均光密度值分析,C組晶狀體平均光密度值高于B組(P0.05)。C組結(jié)果顯示,C3組與C4組平均光密度值較高,晶狀體透明性較高,組間差異無統(tǒng)計(jì)學(xué)意義(P0.05),其它各組間平均光密度值差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。 2.大鼠晶狀體生化指標(biāo)的測(cè)定培養(yǎng)結(jié)束后,C組大鼠晶狀體組織中的GSH、T-AOC、SOD、CAT含量高于B組(P0.05),在C組中,C3組大鼠晶狀體組織中的GSH、T-AOC、CAT活性最高(P0.05), C3組與C4組SOD含量差異無統(tǒng)計(jì)學(xué)意義(P0.05)。C組大鼠晶狀體組織中MDA含量低于B組(P0.05),而C3、C4、C5組組間MDA含量差異無統(tǒng)計(jì)學(xué)意義(P0.05)。 3.光鏡下各組晶狀體組織的細(xì)胞形態(tài)學(xué)觀察HE染色后,光鏡下可見,空白對(duì)照組晶狀體前囊膜連續(xù)光滑,晶狀體上皮細(xì)胞線狀排列,核呈長(zhǎng)桿狀,晶狀體皮質(zhì)纖維層排列規(guī)整。H202處理組晶狀體前囊膜不連續(xù),晶狀體上皮細(xì)胞有增殖,排列紊亂,部分晶狀體上皮細(xì)胞向深層組織移行,細(xì)胞體積變大,胞核多形。MLT10μmol／L組晶狀體前囊膜完整,晶狀體上皮細(xì)胞增殖,形態(tài)呈長(zhǎng)桿狀,排列呈多層,多位于較深層次；MLT30μmol/L組部分晶狀體上皮細(xì)胞增殖,失去原來整齊的單層排列,出現(xiàn)擁擠現(xiàn)象,晶狀體皮質(zhì)纖維層排列規(guī)整；MLT50μmol／L組晶狀體上皮細(xì)胞線狀排列,間隙較一致,胞核呈長(zhǎng)桿狀,形態(tài)較一致。隨著MLT濃度的增加,可見晶狀體上皮細(xì)胞線狀排列,間隙較一致,少量細(xì)胞位于較深層,晶狀體皮質(zhì)纖維層排列緊密而規(guī)整,但是MLT100μmol／L組有囊泡形成。 研究結(jié)論： 在H202存在的條件下,未加入MLT,大鼠晶狀體全混呈白色。在H2O2存在的條件下,加入MLT,大鼠晶狀體的透明度明顯高于H2O2處理組。MLT及其被氧化后的多級(jí)代謝產(chǎn)物具有較強(qiáng)的抗氧化作用,抵抗H2O2對(duì)晶狀體的氧化損傷。本實(shí)驗(yàn)中加入MLT體外培養(yǎng)的大鼠晶狀體GSH、T-AOC、SOD及CAT含量高于H2O2處理組,而MDA含量低于H2O2處理組,說明MLT組抗氧化能力提高。在加入MLT各組間,從MLT 10μmol/L至MLT50μmol/L,隨著濃度的增加,其抗氧化能力越強(qiáng),MLT在濃度為50μmol/L時(shí)即能發(fā)揮有效作用。光鏡下觀察大鼠晶狀體上皮細(xì)胞的形態(tài),H2O2處理組晶狀體前囊膜不完整,晶狀體上皮細(xì)胞排列紊亂,呈多層,細(xì)胞間隙增大,胞核多形；而MLT組上皮細(xì)胞破壞程度比H2O2處理組輕,MLT50μmol/L組晶狀體上皮細(xì)胞呈單層線狀排列,細(xì)胞間隙較一致,胞核呈長(zhǎng)桿狀,形態(tài)更接近空白對(duì)照組,這些實(shí)驗(yàn)結(jié)果說明MLT可提高氧化損傷晶狀體的抗氧化能力,發(fā)揮了很強(qiáng)的保護(hù)晶狀體、維持晶狀體透明性的作用。
[Abstract]:The purpose of the study is:
Cataract is an unanimous blind eye disease, and oxidative damage is an important factor in inducing cataract. Melatonin (MLT) is an indole hormone secreted by the pineal body. It is a strong effective free radical scavenger and antioxidant in the human body. This subject applies melatonin to the lens of rats with oxidative damage in vitro and tests the antioxidant model in vitro. To explore the inhibitory effect of melatonin on the formation of cataract induced by hydrogen peroxide in rats, and to provide a theoretical basis for seeking more ideal antioxidation drugs.
Research methods:
The lens of Wistar rats developed in vitro was divided into three groups according to the random principle: group A was blank control group, group B was hydrogen peroxide (H2O2), and C group was added with different concentrations of MLT on the basis of group B, and the final concentration of MLT was 10 (C1 group), 30 (C2 group), 50 (C3 group), 100 (C4 group), 200 (Group)). The degree of turbidity in the lenses of each group was observed. The average optical density of the black strip under the lens was analyzed by Image-ProPlus (IPP) 6. The protein content of the lens tissue in each group was measured by the enzyme labeling instrument, and the glutathione (GSH), the malondialdehyde (MDA), the total antioxidant energy (T-AOC), the superoxide dismutase (SOD) and the catalase (CAT) content were also measured. The morphology of lens epithelial cells was observed under inverted microscope with HE staining.
The results of the study:
After the observation and quantitative analysis of the degree of lens opacities in 1. rats, the lens under the slit lamp was visible, the lens of the A group remained transparent and the posterior ash line of the lens was clearly visible; the B group was completely mixed with white, and the posterior ash line of the lens was blurred; the transparency of the C group was higher than that of the B group, and the posterior ash line of the lens was blurred in varying degrees. The mean light density value of C group was higher than that of group B (P0.05).C group. The mean light density of C3 group and C4 group was higher, and the transparency of lens was higher. There was no statistical difference between groups (P0.05), and the difference of average light density between the other groups had statistical significance (P0.05).
The content of GSH, T-AOC, SOD, CAT in the lens tissue of group C rats was higher than that of group B (P0.05) after the determination of the biochemical indexes of the lens. In group C, GSH, T-AOC, and CAT activity in the lens tissues of group C3 rats were the highest. In group B (P0.05), there was no significant difference in MDA content between groups C3, C4 and C5 (P0.05).
The morphology of the lens tissue in each group was observed under the 3. light microscope after HE staining. In the blank control group, the anterior capsule of the lens was smooth, the lens epithelial cells were arranged linearly, the nucleus was long rod like, the fibrous layer arranged in the lens cortex was discontinuous in the pre lens capsule of the.H202 treatment group, and the lens epithelial cells proliferated and arranged turbulence. In disorder, the epithelial cells of part of the lens moved to the deep tissue, the cell volume became larger, the nucleus of the nucleus polymorphs.MLT10 Mu mol / L was complete, the epithelial cells of the lens proliferated, the morphology was long rod like, and the arrangement was multi-layer and more in the deeper level, and the MLT30 mu mol/L group partial crystalline epithelial cells proliferated and lost the original neat monolayer arrangement. The fibrous layer of the lens cortex was arranged regularly, and the lens epithelial cells in MLT50 Mu mol / L were arranged linearly, with a relatively uniform space, and the nuclei were long rod-shaped, and the morphology was more consistent. With the increase of MLT concentration, the lens epithelial cells were arranged linearly, the gaps were more consistent, a small number of cells were located in the deeper layer, and the lens cortical fibrous layer arranged. Tight and regular, but vesicles formed in the MLT100 mol / L group.
The conclusions are as follows:
Under the presence of H202, the rat lens was mixed with white without MLT. Under the condition of H2O2, the transparency of the lens was obviously higher than that of the H2O2 treatment group.MLT and the multilevel metabolites after the oxidation, which resisted the oxidative damage of the H2O2 to the lens. In this experiment, MLT in vitro was added to the experiment. The content of GSH, T-AOC, SOD and CAT in cultured rat lens is higher than that in H2O2 treatment group, and the content of MDA is lower than that of H2O2 treatment group. It shows that the antioxidant capacity of MLT group is improved. The antioxidant capacity of MLT group is stronger with the increase of MLT 10 mu mol/L to MLT50. The morphology of the lens epithelial cells in the rats was observed. The anterior capsule of the H2O2 treatment group was incomplete, the lens epithelial cells were arranged in disorder, the lens epithelial cells were multilayered, the cell space was enlarged, and the nucleus was polymorphic. The damage degree of the epithelial cells in the group MLT was lighter than that of the H2O2 treatment group. The epithelial cells in the group of MLT50 mu mol/L were arranged in a single linear arrangement and the cell space was more consistent. The nucleation was long rod like, and the morphology was closer to the blank control group. These results showed that MLT could improve the antioxidant capacity of the oxidative damaged lens, and played a strong role in protecting the lens and maintaining the transparency of the lens.

【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R776.1

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