中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2013年
1期
23-27
,共5页
李爱军%房军%朱秀安%于文贞%狄春辉%杨丽萍
李愛軍%房軍%硃秀安%于文貞%狄春輝%楊麗萍
리애군%방군%주수안%우문정%적춘휘%양려평
遗传性视网膜变性%白细胞介素/白细胞介素-1、受体拮抗剂%细胞凋亡
遺傳性視網膜變性%白細胞介素/白細胞介素-1、受體拮抗劑%細胞凋亡
유전성시망막변성%백세포개소/백세포개소-1、수체길항제%세포조망
Inherited retinal degeneration%Interleukin/interleukin-1,receptor antagonist%Apoptosis
背景 遗传性视网膜变性是主要致盲性眼病之一,包括一系列以慢性进行性视网膜变性为病理特征的异常；白细胞介素-1(IL-1)参与变性与凋亡过程的调节,而白细胞介素-1受体拮抗剂(IL-1 ra)对细胞凋亡的影响了解较少,IL-1 ra是否能阻止视网膜变性值得研究. 目的 研究IL-1 ra对自发性遗传性视网膜变性大鼠视细胞凋亡的影响. 方法 选择SPF级出生后9、15、16、25、30、35、40、50、60 d的RCS大鼠各9只9只眼制备视网膜切片,采用TUNEL凋亡检测试剂盒检测不同年龄大鼠视网膜凋亡细胞.选择出生后15d的RCS大鼠9只,右眼玻璃体腔注射质量浓度1.8 g/L IL-1ra 1μl,左眼玻璃体腔注入PBS 1μl作为对照,分别待大鼠20日龄、25日龄时各重复注射药物1次,30日龄时处死实验大鼠并制备视网膜切片,采用TUNEL凋亡检测试剂盒检测及分析视网膜细胞凋亡情况.采用单因素方差分析比较不同鼠龄视网膜中阳性细胞核面积百分率的差异,采用配对t检验比较大鼠IL-1 ra注射组与PBS注射组视网膜各层厚度的差异. 结果 RCS大鼠视网膜视细胞自出生后第25天出现凋亡,第30 ～35天达高峰,不同鼠龄间大鼠视网膜TUNEL阳性视细胞核面积百分率的差异有统计学意义(F=28.020,P＜0.01).TUNEL凋亡图像分析结果显示,30日龄大鼠IL-1ra注射眼视网膜TUNEL阳性染色弱于PBS注射眼,且阳性染色细胞数少于PBS注射眼；IL-1 ra注射眼大鼠视网膜TUNEL阳性视细胞核总面积及相对面积均明显低于PBS注射眼,差异均有统计学意义[总面积:(223.052±153.092) μm2 vs.(1235.050±359.767) μm2,t=4.370,P＜0.01；相对面积:(2.206±1.531)％ vs.(7.269±1.624)％,t=3.250,P＜0.01].IL-1 ra注射组和PBS注射组视锥与视杆细胞层厚度分别为(15.324±9.035) μm和(49.566±4.605) μm,差异有统计学意义(t=22.674,P＜0.01),但2个组间外核层(即由视锥细胞与视杆细胞的细胞体和细胞核组成)厚度比较差异无统计学意义(t=0.268,P＞0.05).结论 遗传性视网膜变性眼中IL-1对视网膜凋亡发挥一定的调控作用,IL-1 ra对遗传性视网膜变性早期具有潜在的临床治疗价值.
揹景 遺傳性視網膜變性是主要緻盲性眼病之一,包括一繫列以慢性進行性視網膜變性為病理特徵的異常；白細胞介素-1(IL-1)參與變性與凋亡過程的調節,而白細胞介素-1受體拮抗劑(IL-1 ra)對細胞凋亡的影響瞭解較少,IL-1 ra是否能阻止視網膜變性值得研究. 目的 研究IL-1 ra對自髮性遺傳性視網膜變性大鼠視細胞凋亡的影響. 方法 選擇SPF級齣生後9、15、16、25、30、35、40、50、60 d的RCS大鼠各9隻9隻眼製備視網膜切片,採用TUNEL凋亡檢測試劑盒檢測不同年齡大鼠視網膜凋亡細胞.選擇齣生後15d的RCS大鼠9隻,右眼玻璃體腔註射質量濃度1.8 g/L IL-1ra 1μl,左眼玻璃體腔註入PBS 1μl作為對照,分彆待大鼠20日齡、25日齡時各重複註射藥物1次,30日齡時處死實驗大鼠併製備視網膜切片,採用TUNEL凋亡檢測試劑盒檢測及分析視網膜細胞凋亡情況.採用單因素方差分析比較不同鼠齡視網膜中暘性細胞覈麵積百分率的差異,採用配對t檢驗比較大鼠IL-1 ra註射組與PBS註射組視網膜各層厚度的差異. 結果 RCS大鼠視網膜視細胞自齣生後第25天齣現凋亡,第30 ～35天達高峰,不同鼠齡間大鼠視網膜TUNEL暘性視細胞覈麵積百分率的差異有統計學意義(F=28.020,P＜0.01).TUNEL凋亡圖像分析結果顯示,30日齡大鼠IL-1ra註射眼視網膜TUNEL暘性染色弱于PBS註射眼,且暘性染色細胞數少于PBS註射眼；IL-1 ra註射眼大鼠視網膜TUNEL暘性視細胞覈總麵積及相對麵積均明顯低于PBS註射眼,差異均有統計學意義[總麵積:(223.052±153.092) μm2 vs.(1235.050±359.767) μm2,t=4.370,P＜0.01；相對麵積:(2.206±1.531)％ vs.(7.269±1.624)％,t=3.250,P＜0.01].IL-1 ra註射組和PBS註射組視錐與視桿細胞層厚度分彆為(15.324±9.035) μm和(49.566±4.605) μm,差異有統計學意義(t=22.674,P＜0.01),但2箇組間外覈層(即由視錐細胞與視桿細胞的細胞體和細胞覈組成)厚度比較差異無統計學意義(t=0.268,P＞0.05).結論 遺傳性視網膜變性眼中IL-1對視網膜凋亡髮揮一定的調控作用,IL-1 ra對遺傳性視網膜變性早期具有潛在的臨床治療價值.
배경 유전성시망막변성시주요치맹성안병지일,포괄일계렬이만성진행성시망막변성위병리특정적이상；백세포개소-1(IL-1)삼여변성여조망과정적조절,이백세포개소-1수체길항제(IL-1 ra)대세포조망적영향료해교소,IL-1 ra시부능조지시망막변성치득연구. 목적 연구IL-1 ra대자발성유전성시망막변성대서시세포조망적영향. 방법 선택SPF급출생후9、15、16、25、30、35、40、50、60 d적RCS대서각9지9지안제비시망막절편,채용TUNEL조망검측시제합검측불동년령대서시망막조망세포.선택출생후15d적RCS대서9지,우안파리체강주사질량농도1.8 g/L IL-1ra 1μl,좌안파리체강주입PBS 1μl작위대조,분별대대서20일령、25일령시각중복주사약물1차,30일령시처사실험대서병제비시망막절편,채용TUNEL조망검측시제합검측급분석시망막세포조망정황.채용단인소방차분석비교불동서령시망막중양성세포핵면적백분솔적차이,채용배대t검험비교대서IL-1 ra주사조여PBS주사조시망막각층후도적차이. 결과 RCS대서시망막시세포자출생후제25천출현조망,제30 ～35천체고봉,불동서령간대서시망막TUNEL양성시세포핵면적백분솔적차이유통계학의의(F=28.020,P＜0.01).TUNEL조망도상분석결과현시,30일령대서IL-1ra주사안시망막TUNEL양성염색약우PBS주사안,차양성염색세포수소우PBS주사안；IL-1 ra주사안대서시망막TUNEL양성시세포핵총면적급상대면적균명현저우PBS주사안,차이균유통계학의의[총면적:(223.052±153.092) μm2 vs.(1235.050±359.767) μm2,t=4.370,P＜0.01；상대면적:(2.206±1.531)％ vs.(7.269±1.624)％,t=3.250,P＜0.01].IL-1 ra주사조화PBS주사조시추여시간세포층후도분별위(15.324±9.035) μm화(49.566±4.605) μm,차이유통계학의의(t=22.674,P＜0.01),단2개조간외핵층(즉유시추세포여시간세포적세포체화세포핵조성)후도비교차이무통계학의의(t=0.268,P＞0.05).결론 유전성시망막변성안중IL-1대시망막조망발휘일정적조공작용,IL-1 ra대유전성시망막변성조기구유잠재적림상치료개치.
Background Inherited retinal degeneration,one of the major causes of blindness worldwide,comprises a large number of disorders characterized by a slow and progressive retinal degeneration.Interleukin-1 (IL-1)was recognized to be involved in inherited retinal degeneration.Whether IL-1 receptor antagonist (IL-1ra) can arrest retinal degeneration is worthy of investigation.Objective This study was to investigate the effects of IL-1ra on photoreceptor apoptosis in Royal College of Surgeons (RCS) rats.Methods The use of the animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.The SPF RCS rats aged 9,15,16,25,30,35,40,50 and 60 postnatal days were collected,with 9 rats for each age group.Retinal sections were used for the TdT-mediated biotin-dUTP nick-end labeling (TUNEL) cell apoptosis assay.1 μl of IL-1ra (1.8 g/L) was intravitreally injected in the right eyes of 9 RCS rats aged 15 postnatal days and PBS was used in the same way in the fellow eyes.The injection procedure was repeated on the 20 th and 25 th day,respectively.The rats were sacrificed on the 30 th day and retinal sections were prepared for the TUNEL assay.The differences in the percentage of the positive cellular nuclei area among different ages of rats were compared by one-way ANOVA,and the differences in retinal layer thickness between IL-1ra injection group and PBS injection group were assessed by paired t test.Results Photoreceptor apoptosis appeared in 20-day-old RCS rats and progressed and peaked in 30 and 35-day-old rats and then reduced,showing a significant difference among rat of various age groups (F=28.020,P＜0.01).Images from TUNEL assay showed a weaker and less TUNEL staining in the IL-1ra injected eyes than the PBS injected eyes in 30-day-old rats.The total area and relative area of TUNEL positive nuclei were (223.052±153.092) μm2 and (2.206±1.531) ％ in the IL-1ra injected group,and those in PBS injected group were (1235.050±359.767) μm2 and (7.269± 1.624) ％,with a significant difference between them (t =4.370,t=3.250,P＜0.01).The cone and rod thickness was (15.324±9.035) μm in the IL-1ra injected group and (49.566±4.605)μm in the PBS injected group,showing a significant difference (t =22.674,P＜0.01).However,no significant difference was seen in the outer nuclear layer thickness between the two groups (t =0.268,P＞0.05).Conclusions IL-1 participates in the pathogenesis and development of inherited retinal degeneration of RCS rats.The use of IL-1ra might be a potential approach in the treatment of inherited retinal degeneration.
