中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2014年
2期
137-142
,共6页
刘溢%张玉杰%夏丹%姚志峰%袁志兰
劉溢%張玉傑%夏丹%姚誌峰%袁誌蘭
류일%장옥걸%하단%요지봉%원지란
11β羟类固醇脱氢酶1/生理功能%糖皮质激素/不良反应%高眼压/药物诱导%青光眼%受体,皮质类固醇/代谢%动物
11β羥類固醇脫氫酶1/生理功能%糖皮質激素/不良反應%高眼壓/藥物誘導%青光眼%受體,皮質類固醇/代謝%動物
11β간류고순탈경매1/생리공능%당피질격소/불량반응%고안압/약물유도%청광안%수체,피질류고순/대사%동물
11β-Hydroxysteroid dehydrogenase type 1/physiological function%Glucocorticoids/adverse effect%Ocular hypertension/chemically induced%Glaucoma%Receptor,Steroid/metabolism%Animal
背景 糖皮质激素的长期应用可能诱发敏感个体的眼压升高,研究证实11β羟类固醇脱氢酶1(11 β-HSD1)及糖皮质激素受体(GR)、盐皮质激素受体(MR)可能影响房水的产生,这种机制是否与糖皮质激素性高眼压的发生有关目前国内外研究甚少. 目的 检测11β-HSD1及GR、MR在兔糖皮质激素性高眼压模型睫状体组织中的表达变化,探讨其在糖皮质激素性高眼压发病中的作用. 方法 采用随机数字表法将12～16周龄新西兰白兔13只随机分为对照组5只和实验组8只.实验组兔隔日在固定时间及固定地点于左眼球结膜下注射质量分数0.5％地塞米松磷酸钠注射液5 mg(1 ml),注射次日用质量分数0.5％地塞米松滴眼液点眼3次.每次注射后用氧氟沙星滴眼液点眼3次以预防感染,持续2个月；对照组兔以同法注射等容积无菌生理盐水,实验组兔眼压升高至18 mmHg(1 mmHg=0.133 kPa)以上并能持续1周者为造模成功.实验组注射5周后以过量麻醉法处死各组实验兔,分离模型眼睫状体组织,采用免疫组织化学法检测和定位11β-HSD1蛋白在兔模型眼睫状体中的表达；采用逆转录-PCR(RT-PCR)法定量检测11β-HSD1 mRNA、GRmRNA及MR mRNA在实验组兔模型眼睫状体组织中的表达,并与对照组兔眼进行比较. 结果 实验组采用地塞米松局部注射联合点眼后前2周眼压无明显升高,第1周与第2周间眼压差异无统计学意义(q=0.469,P＞0.05)；第3、4、5周眼压逐渐升高,与对照组比较差异均有统计学意义(q=10.535、20.353、28.681,P＜0.01),第5周时达(18.87±0.77) mmHg.免疫组织化学法检测显示,11β-HSD1蛋白在睫状体中呈阳性表达,位于非色素上皮细胞,但模型眼11β-HSD1的阳性染色强度明显弱于对照组.RT-PCR法检测显示,实验组兔模型眼睫状体中MR mRNA、GR mRNA及11β-HSD1 mRNA的相对表达量(以GADPH为内参)分别为2.22±0.78、0.64±0.11、0.47±0.16,而对照组分别为0.94±0.27、1.88±0.74、2.68±1.28,差异均有统计学意义(t=6.070,P=0.004;t5.170,P=0.007；t=5.540,P=0.005).结论 外源性糖皮质激素性高眼压的发病可能与睫状体中MR表达的上调及11β-HSD1和GR表达的下调有关.
揹景 糖皮質激素的長期應用可能誘髮敏感箇體的眼壓升高,研究證實11β羥類固醇脫氫酶1(11 β-HSD1)及糖皮質激素受體(GR)、鹽皮質激素受體(MR)可能影響房水的產生,這種機製是否與糖皮質激素性高眼壓的髮生有關目前國內外研究甚少. 目的 檢測11β-HSD1及GR、MR在兔糖皮質激素性高眼壓模型睫狀體組織中的錶達變化,探討其在糖皮質激素性高眼壓髮病中的作用. 方法 採用隨機數字錶法將12～16週齡新西蘭白兔13隻隨機分為對照組5隻和實驗組8隻.實驗組兔隔日在固定時間及固定地點于左眼毬結膜下註射質量分數0.5％地塞米鬆燐痠鈉註射液5 mg(1 ml),註射次日用質量分數0.5％地塞米鬆滴眼液點眼3次.每次註射後用氧氟沙星滴眼液點眼3次以預防感染,持續2箇月；對照組兔以同法註射等容積無菌生理鹽水,實驗組兔眼壓升高至18 mmHg(1 mmHg=0.133 kPa)以上併能持續1週者為造模成功.實驗組註射5週後以過量痳醉法處死各組實驗兔,分離模型眼睫狀體組織,採用免疫組織化學法檢測和定位11β-HSD1蛋白在兔模型眼睫狀體中的錶達；採用逆轉錄-PCR(RT-PCR)法定量檢測11β-HSD1 mRNA、GRmRNA及MR mRNA在實驗組兔模型眼睫狀體組織中的錶達,併與對照組兔眼進行比較. 結果 實驗組採用地塞米鬆跼部註射聯閤點眼後前2週眼壓無明顯升高,第1週與第2週間眼壓差異無統計學意義(q=0.469,P＞0.05)；第3、4、5週眼壓逐漸升高,與對照組比較差異均有統計學意義(q=10.535、20.353、28.681,P＜0.01),第5週時達(18.87±0.77) mmHg.免疫組織化學法檢測顯示,11β-HSD1蛋白在睫狀體中呈暘性錶達,位于非色素上皮細胞,但模型眼11β-HSD1的暘性染色彊度明顯弱于對照組.RT-PCR法檢測顯示,實驗組兔模型眼睫狀體中MR mRNA、GR mRNA及11β-HSD1 mRNA的相對錶達量(以GADPH為內參)分彆為2.22±0.78、0.64±0.11、0.47±0.16,而對照組分彆為0.94±0.27、1.88±0.74、2.68±1.28,差異均有統計學意義(t=6.070,P=0.004;t5.170,P=0.007；t=5.540,P=0.005).結論 外源性糖皮質激素性高眼壓的髮病可能與睫狀體中MR錶達的上調及11β-HSD1和GR錶達的下調有關.
배경 당피질격소적장기응용가능유발민감개체적안압승고,연구증실11β간류고순탈경매1(11 β-HSD1)급당피질격소수체(GR)、염피질격소수체(MR)가능영향방수적산생,저충궤제시부여당피질격소성고안압적발생유관목전국내외연구심소. 목적 검측11β-HSD1급GR、MR재토당피질격소성고안압모형첩상체조직중적표체변화,탐토기재당피질격소성고안압발병중적작용. 방법 채용수궤수자표법장12～16주령신서란백토13지수궤분위대조조5지화실험조8지.실험조토격일재고정시간급고정지점우좌안구결막하주사질량분수0.5％지새미송린산납주사액5 mg(1 ml),주사차일용질량분수0.5％지새미송적안액점안3차.매차주사후용양불사성적안액점안3차이예방감염,지속2개월；대조조토이동법주사등용적무균생리염수,실험조토안압승고지18 mmHg(1 mmHg=0.133 kPa)이상병능지속1주자위조모성공.실험조주사5주후이과량마취법처사각조실험토,분리모형안첩상체조직,채용면역조직화학법검측화정위11β-HSD1단백재토모형안첩상체중적표체；채용역전록-PCR(RT-PCR)법정량검측11β-HSD1 mRNA、GRmRNA급MR mRNA재실험조토모형안첩상체조직중적표체,병여대조조토안진행비교. 결과 실험조채용지새미송국부주사연합점안후전2주안압무명현승고,제1주여제2주간안압차이무통계학의의(q=0.469,P＞0.05)；제3、4、5주안압축점승고,여대조조비교차이균유통계학의의(q=10.535、20.353、28.681,P＜0.01),제5주시체(18.87±0.77) mmHg.면역조직화학법검측현시,11β-HSD1단백재첩상체중정양성표체,위우비색소상피세포,단모형안11β-HSD1적양성염색강도명현약우대조조.RT-PCR법검측현시,실험조토모형안첩상체중MR mRNA、GR mRNA급11β-HSD1 mRNA적상대표체량(이GADPH위내삼)분별위2.22±0.78、0.64±0.11、0.47±0.16,이대조조분별위0.94±0.27、1.88±0.74、2.68±1.28,차이균유통계학의의(t=6.070,P=0.004;t5.170,P=0.007；t=5.540,P=0.005).결론 외원성당피질격소성고안압적발병가능여첩상체중MR표체적상조급11β-HSD1화GR표체적하조유관.
Background Long-term administration of glucocorticoid drugs induces ocular hypertension in susceptible individuals probably.It has been verified that 1 1β-hydroxysteroid dehydrogenase type 1 (11β-HSD1),glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) can affect the generating of aqueous humor,but how they play the role in glucocorticoid-induced ocular hypertension is unclear.Objective This study was to investigate the relationship of expressions of 11β-HSD1 and steroids receptors in ciliary body and steroid-induced ocular hypertension.Methods Thirteen 12-16 week-old New Zealand albino rabbits were randomized to control group (5 rabbits) and experimental group (8 rabbits).Steroid-induced glaucoma models were induced by administration of subconjunctival injection of 5 mg dexamethasone solution(1 ml) and 0.5％ dexamethasone eye drops on alternate days in the left eyes for consecutive two months in the experimental group,and the equal volume of sterile normal saline solution was used in the same way in the control group.The successful criteria of model eyes was defined as rising of intraocular pressure (IOP) to ≥ 18 mmHg for over one week.Then,the animals were sacrificed by excessive anesthesia and the ciliary tissues were isolated for the assay of expressions of 1 1β-HSD1 protein by immunochemistry,and the expressions of 11β-HSD1 mRNA,GR mRNA and MR mRNA in ciliary body were semi-quantitatively detected by reverse transcription-PCR (RT-PCR).The experimental results were compared between the two groups.Results The IOP was normal in the first two weeks after administration of drugs,and no significant difference was found in IOP between the first week and the second week in the experimental group (q =0.469,P ＞0.05).From 3 through 5 weeks after injection,the IOP was gradually elevated,with the highest value of (18.87±0.77) mmHg in the fifth week.Significant differences were seen between the two groups at mentioned-above time points (q =10.535,20.353,28.681,all at P ＜ 0.01).11β-HSD1 protein was positively expressed in nonpigmented epithelial cells of ciliary tissue of rabbits in both groups,however,the expression intensity was weaker in the experimental group compared with the control group.The relative expressional values of MR mRNA,GR mRNA and 11β-HSD1 mRNA in the ciliary tissue were 2.22±0.78,0.64±0.11 and 0.47±0.16 in the experimental group,and those in the control group were 0.94±0.27,1.88±0.74 and 2.68±1.28,with significant differences between the two groups (t =6.070,P =0.004 ; t =5.170,P =0.007 ; t =5.540,P =0.005).Conclusions Corticosteroidinduced glaucoma probably is associated with the up-regulation of MR level and down-regulations of GR and 11β-HSD1 in ciliary body.
