中华眼视光学与视觉科学杂志
中華眼視光學與視覺科學雜誌
중화안시광학여시각과학잡지
CHINESE JOURNAL OF OPTOMETRY OPHTHALMOLOGY AND VISUAL SCIENCE
2012年
5期
276-280
,共5页
蛋白硫酸软骨素类%碘酸盐%视网膜炎%色素性%小神经胶质细胞
蛋白硫痠軟骨素類%碘痠鹽%視網膜炎%色素性%小神經膠質細胞
단백류산연골소류%전산염%시망막염%색소성%소신경효질세포
Proteochondroitin Sulfates%Iodates%Retinitis%pigmentosa%Microglia
目的 研究硫酸软骨素蛋白多糖(CSPGs)在碘酸钠(NaIO3)诱导的视网膜色素上皮(RPE)细胞变性大鼠中的表达情况.方法 实验研究.24只Sprauge-Dawley(SD)大鼠随机分为4组,正常对照组、造模7d组、造模14 d组、造模28 d组,每组6只.造模组腹腔注射3％ NaIO3(100 mg/kg);取眼球做病理检查,苏木素-伊红(HE)染色及细胞凋亡检测,验证视网膜变性动物模型的建立;免疫荧光法观察视网膜变性大鼠视网膜上CSPGs表达的时空规律；逆转录-聚合酶链反应(RT-PCR)法检测CSPGs多功能蛋白聚糖(Versican) mRNA的表达情况；免疫组织化学法观察视网膜上小胶质细胞表达的巨噬细胞特异性抗原CD68的分布特点.多组比较采用单因素方差分析,两样本间比较采用独立样本t检验.结果 注射NaIO3后,模型大鼠视网膜出现变性改变,且光感受器发牛渐进性凋亡,造模7、14、28 d组凋亡率分别为(21.0±3.5)％、(32.3±2.3)％、(41.7±2.6)％,各组间差异有统计学意义(F=205.27,P＜0.01).造模7d组,CS-56、CD68在脉络膜、视锥视杆层、内核层、节细胞层表达;造模14 d组,CS-56、CD68进一步出现在外核层;造模28 d组,CSPGs继续迁移到外丛状层.随造模时间延长,各层荧光表达逐渐增强,CD68在视网膜各层的表达也明显增多,强度分别为:1.33±0.52、2.67±0.82、4.00±1.10和7.17±1.33,各组间差异有统计学意义(F=38.45,P＜0.01).造模组Versican mRNA表达(1.02±0.06)显著高于正常对照组(0.23±0.02),差异有统计学意义(t=-26.05,P＜0.01).结论 在碘酸钠诱导的视网膜变性动物模型中,随时间延长CSPGs表达范围扩大,表达量增加,并与小胶质细胞分布在大致相同区域,提示CSPGs可能来源于小胶质细胞.
目的 研究硫痠軟骨素蛋白多糖(CSPGs)在碘痠鈉(NaIO3)誘導的視網膜色素上皮(RPE)細胞變性大鼠中的錶達情況.方法 實驗研究.24隻Sprauge-Dawley(SD)大鼠隨機分為4組,正常對照組、造模7d組、造模14 d組、造模28 d組,每組6隻.造模組腹腔註射3％ NaIO3(100 mg/kg);取眼毬做病理檢查,囌木素-伊紅(HE)染色及細胞凋亡檢測,驗證視網膜變性動物模型的建立;免疫熒光法觀察視網膜變性大鼠視網膜上CSPGs錶達的時空規律；逆轉錄-聚閤酶鏈反應(RT-PCR)法檢測CSPGs多功能蛋白聚糖(Versican) mRNA的錶達情況；免疫組織化學法觀察視網膜上小膠質細胞錶達的巨噬細胞特異性抗原CD68的分佈特點.多組比較採用單因素方差分析,兩樣本間比較採用獨立樣本t檢驗.結果 註射NaIO3後,模型大鼠視網膜齣現變性改變,且光感受器髮牛漸進性凋亡,造模7、14、28 d組凋亡率分彆為(21.0±3.5)％、(32.3±2.3)％、(41.7±2.6)％,各組間差異有統計學意義(F=205.27,P＜0.01).造模7d組,CS-56、CD68在脈絡膜、視錐視桿層、內覈層、節細胞層錶達;造模14 d組,CS-56、CD68進一步齣現在外覈層;造模28 d組,CSPGs繼續遷移到外叢狀層.隨造模時間延長,各層熒光錶達逐漸增彊,CD68在視網膜各層的錶達也明顯增多,彊度分彆為:1.33±0.52、2.67±0.82、4.00±1.10和7.17±1.33,各組間差異有統計學意義(F=38.45,P＜0.01).造模組Versican mRNA錶達(1.02±0.06)顯著高于正常對照組(0.23±0.02),差異有統計學意義(t=-26.05,P＜0.01).結論 在碘痠鈉誘導的視網膜變性動物模型中,隨時間延長CSPGs錶達範圍擴大,錶達量增加,併與小膠質細胞分佈在大緻相同區域,提示CSPGs可能來源于小膠質細胞.
목적 연구류산연골소단백다당(CSPGs)재전산납(NaIO3)유도적시망막색소상피(RPE)세포변성대서중적표체정황.방법 실험연구.24지Sprauge-Dawley(SD)대서수궤분위4조,정상대조조、조모7d조、조모14 d조、조모28 d조,매조6지.조모조복강주사3％ NaIO3(100 mg/kg);취안구주병리검사,소목소-이홍(HE)염색급세포조망검측,험증시망막변성동물모형적건립;면역형광법관찰시망막변성대서시망막상CSPGs표체적시공규률；역전록-취합매련반응(RT-PCR)법검측CSPGs다공능단백취당(Versican) mRNA적표체정황；면역조직화학법관찰시망막상소효질세포표체적거서세포특이성항원CD68적분포특점.다조비교채용단인소방차분석,량양본간비교채용독립양본t검험.결과 주사NaIO3후,모형대서시망막출현변성개변,차광감수기발우점진성조망,조모7、14、28 d조조망솔분별위(21.0±3.5)％、(32.3±2.3)％、(41.7±2.6)％,각조간차이유통계학의의(F=205.27,P＜0.01).조모7d조,CS-56、CD68재맥락막、시추시간층、내핵층、절세포층표체;조모14 d조,CS-56、CD68진일보출현재외핵층;조모28 d조,CSPGs계속천이도외총상층.수조모시간연장,각층형광표체축점증강,CD68재시망막각층적표체야명현증다,강도분별위:1.33±0.52、2.67±0.82、4.00±1.10화7.17±1.33,각조간차이유통계학의의(F=38.45,P＜0.01).조모조Versican mRNA표체(1.02±0.06)현저고우정상대조조(0.23±0.02),차이유통계학의의(t=-26.05,P＜0.01).결론 재전산납유도적시망막변성동물모형중,수시간연장CSPGs표체범위확대,표체량증가,병여소효질세포분포재대치상동구역,제시CSPGs가능래원우소효질세포.
Objective To study the expression of chondroitin sulfate proteoglycans (CSPGs) in retinal pigment epithelium cell degeneration rat induced by sodium iodate (NaIO3) intraperitoneal injection.Methods In this experimental study,a total of 24 Sprague-Dawley (SD) rats were used.And divided into control group,7 days group,14 days group and 28 days group,with 6 rats in each group.NaIO3 (3％,100 mg/kg) was intraperitoneally injected in 18 rats to establish the retinal degeneration models.HE and Tunel assay were performed to evaluate the retinal degeneration.The expression of CSPGs in rat retinas was detected by immunofluorescence; the expression of Versican mRNA was analyzed by retrovirus-polymerase chain reaction (RT-PCR).The macrophage-specific antigen CD68,secreted by microglia,was detected by an immunohistochemical method.The data was statistically analyzed by one-way ANOVA and independent samples t test.Results HE showed morphological retinal degeneration in rats after NaIO3 injection.The apoptosis of photoreceptors appeared more severe with the development of retinal degeneration in rat retinas after NaIO3 injection.The apoptosis rates were (21.0±3.5)％,(323±2.3)％ and (41.7±2.6)％ in 7 days.14 days and 28 days groups,respectively.after NaIO3 injection,with statistically significant differences between each group (F=205.27,P＜0.01).CS-56 and CD68 appeared in the choroid,photoreceptor outer segment debris zone (DZ),the internuclear layer (INL) and the ganglion cell layer (GCL) 7 days after NaIO3 injection and spread to the outer nuclear layer (ONL) 14 days after NaIO3 injection.Twenty-eight days after NaIO3 injection,CSPGs continued to expand to the outer plexiform layers (OPL).With the passage of time after NaIO3 injection,CS-56 fluorescence and CD68 intensity became stronger in each layer.CD68 intensity was 1.33±0.52,2.67±0.82,4.00±1.10 and 7.17±1.33 in normal rat retinas,7 days,14 days and 28 days after NaIO3 injection,and there were statistically significant differences between each group (F=38.45,P＜0.01).The expression of Versican mRNA,the N-terminal fragment of CSPGs,in the retinal degeneration animals was 1.02±0.06,which was significantly higher than in the control rats (0.23±0.02) (t=-26.05,P＜0.01).Conclusion With the passage of time after NaIO3 injection,the range of CSPGs continued to expand and the expression intensity became stronger in rat retinas.The same distribution of CSPGs and microglia suggested that microglia constitute a source of CSPGs in the degenerating rat retina.
