替普瑞酮對急性高眼壓大鼠視神經(jīng)保護作用的實驗研究
發(fā)布時間：2018-10-24 09:03
【摘要】： 目的青光眼目前已成為世界上最主要的致盲眼病之一,其典型表現(xiàn)是視神經(jīng)乳頭的凹陷性萎縮和視野的特征性缺損縮小。視網(wǎng)膜神經(jīng)節(jié)細胞(retinal ganglion cells, RGCs)凋亡,一種高度調控的細胞死亡,是青光眼視神經(jīng)損傷的最終共同通路。用藥物或其他方法干預,使那些未受損的,僅部分受損的,或正處于毒性內環(huán)境中臨近死亡的視網(wǎng)膜細胞得以存活或延長生存時間具有重大意義,也成為當前研究的熱點。熱休克蛋白70(heat shock protein 70,HSP70)是組織缺血損失時敏感而可靠的指標,HSP70表達的增強對神經(jīng)組織具有保護作用。本實驗通過制作急性高眼壓大鼠模型,探討替普瑞酮(geranylgeranylacetone, GGA)對大鼠視網(wǎng)膜HSP70表達的影響,及其對急性高眼壓大鼠視神經(jīng)的保護作用。 方法健康SD大鼠60只隨機分為3組,分為健康對照組大鼠20只(A組)、急性高眼壓對照組20只(B組)和GGA治療組20只(C組),A組和B組給予生理鹽水灌胃,C組給予替普瑞酮灌胃,一周后B組和C組采用前房加壓法制作大鼠急性高眼壓模型,A組不加任何干預措施。術后6h、24h、48h、72h處死動物,完整取出眼球,放人事先編號盛有10%中性甲醛的瓶中保存。石蠟包埋,制作切片。HE染色,光鏡下觀察再灌注后不同時間段各組鼠視網(wǎng)膜組織結構變化,采用免疫組織化學法檢測大鼠視網(wǎng)膜HSP70和凋亡蛋白酶-3(Cysteine-containing aspartate-specific proteases-3, caspase-3)的表達,陽性結果為胞漿或胞核呈黃色或棕黃色染色。計數(shù),采用SPSS16.0統(tǒng)計軟件處理,采用獨立樣本t檢驗,P＜0.05表示差異有統(tǒng)計學意義。 結果術后A組均為正常視網(wǎng)膜結構。C組大鼠視網(wǎng)膜水腫較B組輕,各時間段C組大鼠視網(wǎng)膜內層厚度均較B組厚。B組HSP70在術后6h即有表達,隨時間段延長表達逐漸增加,至再灌注后24 h達到高峰,之后漸下降。C組HSP70表達較之明顯增強,差異有統(tǒng)計學意義(P＜0.05)。B組caspase-3于術后6h開始表達,并逐漸遞增,24 h達到高峰,48h開始下降,72h僅有少量表達。C組caspase-3表達較之明顯減弱,差異有統(tǒng)計學意義(P＜0.05)。A組幾乎無HSP70和caspase-3的表達。 結論急性高眼壓可以造成視神經(jīng)缺血,引起視網(wǎng)膜水腫和RGCs凋亡,同時能誘導視網(wǎng)膜HSP70的表達。替普瑞酮能誘導視網(wǎng)膜HSP70的表達增強,對急性高眼壓造成的視神經(jīng)損傷起到保護作用。
[Abstract]:Objective at present, glaucoma has become one of the most important blinding eye diseases in the world. The typical manifestation of glaucoma is the concave atrophy of optic nipple and the reduction of characteristic defect of visual field. Retinal ganglion (retinal ganglion cells, RGCs) apoptosis, a highly regulated cell death, is the ultimate common pathway of glaucoma optic nerve injury. It is of great significance to intervene with drugs or other methods to make retinal cells that are not damaged, only partially damaged, or near death in toxic environment survive or prolong their survival time, which has also become a hot topic in current research. Heat shock protein 70 (HSP70) is a sensitive and reliable marker of ischemic tissue loss. The enhanced expression of HSP70 has protective effect on nerve tissue. To investigate the effect of tipranone (geranylgeranylacetone, GGA) on the expression of HSP70 in rat retina and its protective effect on optic nerve in rats with acute intraocular hypertension. Methods Sixty healthy SD rats were randomly divided into 3 groups: control group (n = 20), acute intraocular hypertension group (n = 20) and GGA treatment group (n = 20). One week later, group B and group C used anterior chamber pressure method to make acute intraocular hypertension model in rats, group A did not add any intervention measures. The animals were killed at 24 hours and 48 hours after operation. The eyeballs were taken out and stored in a bottle containing 10% neutral formaldehyde. The changes of retinal tissue structure were observed under light microscope at different time after reperfusion. The expression of HSP70 and apoptotic protease 3 (Cysteine-containing aspartate-specific proteases-3, caspase-3) in rat retina were detected by immunohistochemical method. The positive results were yellow or brown staining in cytoplasm or nucleus. The count was processed by SPSS16.0 statistical software, and the independent sample t test was used. The difference was statistically significant (P < 0. 05). Results the retinal edema in group C was lighter than that in group B. the thickness of inner retinal layer in group C was thicker than that in group B. the expression of HSP70 in group B was observed 6 hours after operation and increased gradually with the extension of time. The expression of HSP70 in group C was significantly higher than that in group C (P < 0. 05). The expression of caspase-3 in group C was significantly higher than that in group C (P < 0. 05). The expression of caspase-3 began at 6 hours after reperfusion (P < 0. 05). The expression of caspase-3 in group C was significantly lower than that in group C (P < 0. 05, P < 0. 05). The expression of HSP70 and caspase-3 was almost no in group C (P < 0. 05). Conclusion Acute intraocular pressure can induce optic nerve ischemia, retinal edema and RGCs apoptosis, and induce the expression of retinal HSP70. Tipranone can induce increased expression of HSP70 in retina and protect optic nerve injury caused by acute intraocular pressure.
【學位授予單位】：大連醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R775
[Abstract]:Objective at present, glaucoma has become one of the most important blinding eye diseases in the world. The typical manifestation of glaucoma is the concave atrophy of optic nipple and the reduction of characteristic defect of visual field. Retinal ganglion (retinal ganglion cells, RGCs) apoptosis, a highly regulated cell death, is the ultimate common pathway of glaucoma optic nerve injury. It is of great significance to intervene with drugs or other methods to make retinal cells that are not damaged, only partially damaged, or near death in toxic environment survive or prolong their survival time, which has also become a hot topic in current research. Heat shock protein 70 (HSP70) is a sensitive and reliable marker of ischemic tissue loss. The enhanced expression of HSP70 has protective effect on nerve tissue. To investigate the effect of tipranone (geranylgeranylacetone, GGA) on the expression of HSP70 in rat retina and its protective effect on optic nerve in rats with acute intraocular hypertension. Methods Sixty healthy SD rats were randomly divided into 3 groups: control group (n = 20), acute intraocular hypertension group (n = 20) and GGA treatment group (n = 20). One week later, group B and group C used anterior chamber pressure method to make acute intraocular hypertension model in rats, group A did not add any intervention measures. The animals were killed at 24 hours and 48 hours after operation. The eyeballs were taken out and stored in a bottle containing 10% neutral formaldehyde. The changes of retinal tissue structure were observed under light microscope at different time after reperfusion. The expression of HSP70 and apoptotic protease 3 (Cysteine-containing aspartate-specific proteases-3, caspase-3) in rat retina were detected by immunohistochemical method. The positive results were yellow or brown staining in cytoplasm or nucleus. The count was processed by SPSS16.0 statistical software, and the independent sample t test was used. The difference was statistically significant (P < 0. 05). Results the retinal edema in group C was lighter than that in group B. the thickness of inner retinal layer in group C was thicker than that in group B. the expression of HSP70 in group B was observed 6 hours after operation and increased gradually with the extension of time. The expression of HSP70 in group C was significantly higher than that in group C (P < 0. 05). The expression of caspase-3 in group C was significantly higher than that in group C (P < 0. 05). The expression of caspase-3 began at 6 hours after reperfusion (P < 0. 05). The expression of caspase-3 in group C was significantly lower than that in group C (P < 0. 05, P < 0. 05). The expression of HSP70 and caspase-3 was almost no in group C (P < 0. 05). Conclusion Acute intraocular pressure can induce optic nerve ischemia, retinal edema and RGCs apoptosis, and induce the expression of retinal HSP70. Tipranone can induce increased expression of HSP70 in retina and protect optic nerve injury caused by acute intraocular pressure.
【學位授予單位】：大連醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R775
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