發(fā)布時間：2018-06-18 05:10

  本文選題：硫酸鋅 + 熱休克蛋白70��； 參考：《青島大學(xué)》2012年碩士論文

【摘要】：目的探討大鼠視網(wǎng)膜興奮損傷中HSP70與TNF-α表達(dá)的相關(guān)變化。 方法健康成年清潔級Wistar大鼠50只隨機(jī)分為三組,正常對照組(5只)、陽性對照組即NMDA玻璃體腔注射組(20只)、實(shí)驗(yàn)組即NMDA玻璃體腔注射組+硫酸鋅腹腔注射組(25只)。實(shí)驗(yàn)組腹腔注射硫酸鋅(24.6mg/Kg),10h后隨即處死5只。14h后實(shí)驗(yàn)組和陽性對照組均做單眼的玻璃體腔注射NMDA (2μl 80mmol/l),并做好標(biāo)記。NMDA注射后2h,8h,16h,24h,實(shí)驗(yàn)組和陽性組各組均隨機(jī)處死5只大鼠。正常對照組不做任何處理,可以于任何時間點(diǎn)處死。HE染色光鏡下觀察各組視網(wǎng)膜組織的形態(tài)學(xué)變化,實(shí)時定量RT-PCR方法觀察視網(wǎng)膜TNF-αmRNA和HSP70mRNA表達(dá)的變化。 結(jié)果1.HE染色：正常對照組大鼠視網(wǎng)膜組織結(jié)構(gòu)層次清楚,染色均勻,細(xì)胞形態(tài)規(guī)則。陽性對照組造模后2h,神經(jīng)節(jié)細(xì)胞開始水腫；造模后8h,神經(jīng)節(jié)細(xì)胞水腫最嚴(yán)重,其內(nèi)可見空泡,視網(wǎng)膜增厚；造模后16h,神經(jīng)節(jié)細(xì)胞水腫較前減輕,排列疏松,形態(tài)不規(guī)則,可見核固縮,數(shù)量減少；造模后24h,神經(jīng)節(jié)細(xì)胞數(shù)目減少排列紊亂,核固縮明顯,神經(jīng)纖維層、神經(jīng)節(jié)細(xì)胞層變薄。各時間點(diǎn)實(shí)驗(yàn)組與陽性對照組相比,視網(wǎng)膜的厚度較厚,存活的視網(wǎng)膜神經(jīng)節(jié)細(xì)胞的數(shù)量較多,損傷均輕于陽性對照組。2.熒光定量RT-PCR:正常對照組大鼠視網(wǎng)膜未見TNF-αmRNA表達(dá),可見HSP70微量表達(dá)。陽性對照組中NMDA注射后2h, TNF-αmRNA水平較正常對照組上調(diào)(P0.05),注射后8h, TNF-αmRNA水平達(dá)高峰(P0.05),注射后16h表達(dá)量降低,但仍高于對照組(P0.05),注射后24h, TNF-αmRNA表達(dá)量與16h無差別(p0.05),較正常對照組增多(P0.05); NMDA注射后2h, HSP70 mRNA水平較正常對照組上調(diào)(P0.05),注射后8h, HSP70 mRNA水平明顯上調(diào)(P0.05),注射后16h表達(dá)量達(dá)高峰(P0.05),注射后24h, HSP70 mRNA表達(dá)量降低,較正常對照組增多(P0.05)。實(shí)驗(yàn)組中硫酸鋅注射后10h TNF-αmRNA未見表達(dá),NMDA注射后2h,TNF-αmRNA水平較正常對照組上調(diào)(P0.05),較陽性對照組下降(P0.05),注射后8h,TNF-αmRNA水平達(dá)高峰(P0.05),較陽性對照組下降(P0.05),注射后16h表達(dá)量降低,但仍高于對照組(P0.05),較陽性對照組下降(P0.05),注射后24h,TNF-αmRNA表達(dá)量與16h無差別(p0.05),較陽性對照組下降(P0.05),較正常對照組增多(P0.05)；硫酸鋅注射后10h HSP70 mRNA水平較正常對照組上調(diào)(P0.05),NMDA注射后2h,HSP70 mRNA水平明顯上調(diào)(P0.05),較陽性對照組增加(P0.05),注射后8h, HSP70 mRNA水平達(dá)高峰,與2h的無差別(p0.05),較陽性對照組增加(P0.05),注射后16h,HSP70 mRNA水平與8h的無差別(p0.05),較陽性對照組增加(P＜0.05),注射后24h,HSP70 mRNA水平下降,較陽性對照組增加(P0.05)。 結(jié)論在視網(wǎng)膜興奮損傷中,硫酸鋅可增加HSP70的表達(dá)量,降低TNF-α的表達(dá)量,隨著HSP70的表達(dá)量的增加,TNF-α表達(dá)量下降,表明HSP70對TNF-α的表達(dá)有抑制作用。
[Abstract]:Objective to investigate the correlation between HSP70 and TNF- 偽 expression in rat retina excitatory injury. Methods Fifty healthy adult clean grade Wistar rats were randomly divided into three groups: normal control group (n = 5), NMDA vitreous injection group (n = 20) and zinc sulfate injection group (n = 25). After 10 hours of intraperitoneal injection of zinc sulfate 24.6 mg 路kg ~ (-1) 路L ~ (-1) 路L ~ (-1), 5 rats in the experimental group and the positive control group were killed immediately after 10 h. All the experimental group and the positive control group were given intravitreal injection of NMDA 2 渭 l 80 mmol 路L ~ (-1), and then 2 h, 8 h and 16 h ~ 24 h after injection of .NMDA, 5 rats were randomly killed in the experimental group and the positive group. The normal control group, without any treatment, could be killed at any time. The morphological changes of retinal tissue were observed under light microscope with HE staining, and the expression of TNF- 偽 mRNA and HSP70 mRNA in retina were observed by real-time quantitative RT-PCR. Results 1. HE staining: the retinal tissue structure of normal control group was clear, stained evenly, and the cell morphology was regular. 2. In the positive control group, the ganglion cells began to edema at 2 h after modeling, the ganglion cells showed the most severe edema at 8 h after modeling, with vacuoles and thickened retina, 16 h after modeling, the edema of ganglion cells decreased, the arrangement of ganglion cells was loose, and the shape was irregular. The number of ganglion cells decreased and the number of ganglion cells decreased. The thickness of retina was thicker and the number of surviving retinal ganglion cells was more in the experimental group than in the positive control group at each time point, and the damage was lighter than that in the positive control group. Fluorescence quantitative RT-PCR: no expression of TNF- 偽 mRNA was found in the retina of normal control group, but HSP70 mRNA was detected in the retina. The level of TNF- 偽 mRNA was up-regulated at 2 h after NMDA injection in the positive control group compared with that in the normal control group. The level of TNF- 偽 mRNA reached the peak at 8 h after injection and decreased at 16 h after injection. However, the expression of TNF- 偽 mRNA was still higher than that of the control group at 24 h after injection, and there was no difference between the expression of TNF- 偽 mRNA and that of the control group at 16 h, which was higher than that in the normal control group at 2 h after NMDA injection, and the level of HSP70 mRNA was up-regulated at 2 h after NMDA injection as compared with that in the control group. At 8 h after injection, the mRNA level of HSP70 increased significantly, and the expression of HSP70 mRNA reached to P0.05% at 16 h after injection. The expression of HSP70 mRNA decreased 24 hours after injection. Compared with the normal control group, the number of P0.05 was increased. There was no expression of TNF- 偽 mRNA in the experimental group at 10 h after injection of zinc sulfate. The expression of TNF- 偽 mRNA was higher than that of the normal control group at 2 h after injection, and decreased significantly than that in the positive control group. The level of TNF- 偽 mRNA reached the peak level at 8 h after injection and decreased at 16 h after injection, and decreased at 16 h after injection. However, the expression of TNF- 偽 mRNA was still higher than that of the control group (P 0.05) and decreased than that of the positive control group. The expression of TNF- 偽 mRNA at 24 h after injection was not significantly different from that of the control group at 16 h (P 0.05), but it was lower than that in the positive control group (P 0.05) and increased than that in the normal control group (P 0.05). HSP70 mRNA level was significantly up-regulated at 10 h after injection of zinc sulfate compared with that in normal control group. HSP70 mRNA level in HSP70 was significantly up-regulated at 2 h after injection of P0.05N MDA, and increased significantly than that in positive control group. The HSP70 mRNA level reached its peak at 8 h after injection. The level of HSP70 mRNA was increased at 16 h after injection and the level of HSP70 mRNA at 8 h was similar to that in the positive control group (P < 0.05). The level of HSP70 mRNA decreased 24 hours after injection and increased P0.05 compared with the positive control group at 24 h after injection. Conclusion Zinc sulfate can increase the expression of HSP70 and decrease the expression of TNF- 偽 in retina excitatory injury. With the increase of HSP70, the expression of TNF- 偽 decreases, indicating that HSP70 can inhibit the expression of TNF- 偽.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R774.1

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 邢業(yè)嬌;替普瑞酮對大鼠視網(wǎng)膜興奮性損傷的保護(hù)作用[D];青島大學(xué);2013年

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本文編號：2034249