CAJ | 학술논문

目的：探讨洛美利嗪（lomerizine，LOM）对糖尿病早期大鼠视网膜神经节细胞（ retinal ganglion cells ，RGCs）凋亡的保护作用及其机制。 　　方法：SD大鼠随机分为对照组（ CON ）、糖尿病组（ DM ）及洛美利嗪组（ LOM），每组40只大鼠。 DM和LOM组链脲佐菌素（STZ）按60mg／kg一次性腹腔注射诱导糖尿病模型，CON 组给予等量无菌柠檬酸钠溶液腹腔注射。LOM组于糖尿病鼠模型成模后，每日予LOM 60 mg／kg灌胃，CON组和DM组采用等量生理盐水灌胃。于第4，8，12 wk分别行HE、透射电镜、TUNEL检测RGCs的凋亡情况，同时采用激光共聚焦显微镜检测RGCs内钙离子的浓度。 　　结果：（1）形态学观察：随病程延长，逐渐出现RGCs数量减少、细胞排列紊乱的病理改变。透射电镜下可见DM组RGCs出现不同阶段的凋亡征象，且随病程延长逐渐加重。 LOM组与同期DM组对比，在第8，12 wk时RGCs凋亡征象减弱。（2） TUNEL检测：CON组大鼠视网膜神经节细胞层未见凋亡细胞。 DM组4 wk时视网膜神经节细胞层偶见TUNEL 阳性细胞，并随病程延长逐渐增多，凋亡指数与同期CON组比较明显升高，差异有统计学意义（ P＜0．01）。 LOM组8，12 wk时，与同期DM组比较，染色阳性的RGCs明显减少，凋亡指数明显下降，差异显著（P＜0．01）。（3）激光共聚焦显微镜钙离子浓度检测：DM组与同期CON组比较：DM组8，12 wk 的RGCs内钙离子荧光染色强度明显升高，有显著差异（ P＜0．01）。LOM组与同期DM组比较：LOM组8，12 wk的RGCs内钙离子荧光染色强度明显下降，差异有统计学意义（P＜0．01）。 　　结论：LOM对糖尿病大鼠早期 RGCs 的凋亡具有保护作用。
목적：탐토락미리진（lomerizine，LOM）대당뇨병조기대서시망막신경절세포（ retinal ganglion cells ，RGCs）조망적보호작용급기궤제。 　　방법：SD대서수궤분위대조조（ CON ）、당뇨병조（ DM ）급락미리진조（ LOM），매조40지대서。 DM화LOM조련뇨좌균소（STZ）안60mg／kg일차성복강주사유도당뇨병모형，CON 조급여등량무균저몽산납용액복강주사。LOM조우당뇨병서모형성모후，매일여LOM 60 mg／kg관위，CON조화DM조채용등량생리염수관위。우제4，8，12 wk분별행HE、투사전경、TUNEL검측RGCs적조망정황，동시채용격광공취초현미경검측RGCs내개리자적농도。 　　결과：（1）형태학관찰：수병정연장，축점출현RGCs수량감소、세포배렬문란적병리개변。투사전경하가견DM조RGCs출현불동계단적조망정상，차수병정연장축점가중。 LOM조여동기DM조대비，재제8，12 wk시RGCs조망정상감약。（2） TUNEL검측：CON조대서시망막신경절세포층미견조망세포。 DM조4 wk시시망막신경절세포층우견TUNEL 양성세포，병수병정연장축점증다，조망지수여동기CON조비교명현승고，차이유통계학의의（ P＜0．01）。 LOM조8，12 wk시，여동기DM조비교，염색양성적RGCs명현감소，조망지수명현하강，차이현저（P＜0．01）。（3）격광공취초현미경개리자농도검측：DM조여동기CON조비교：DM조8，12 wk 적RGCs내개리자형광염색강도명현승고，유현저차이（ P＜0．01）。LOM조여동기DM조비교：LOM조8，12 wk적RGCs내개리자형광염색강도명현하강，차이유통계학의의（P＜0．01）。 　　결론：LOM대당뇨병대서조기 RGCs 적조망구유보호작용。
AIM: To investi gate the neuroprotection of lomerizine (LOM) on retinal ganglion cells(RGCs) in early diabetic rats and their mechanism . METHODS: Adult male Spraque -Dawley ( SD ) rats were randomly divided into control ( CON ) , diabetes mellitus ( DM ) , lomeri zine ( LOM ) group, and each group had 40 rats.Diabetes rat model was induced by Streptozotocin ( STZ ) of intraperitoneal in jection of 60 mg/kg of disposable.In LOM group, after model was established, rats were lavaged LOM by the dosage of 60mg/k g daily, the CON group and DM group were given the same dosage sodium chloride.In 4 th, 8 th, 12 th wk, RGCs'apoptosis were det ected by HE, TUNEL, transmission electron microscope, and TUNEL and laser confocal microscope detection was used to test the calcium ion concentration. RESULTS: Morphological observation: with the extension of the DM, RGCs decreased gradually and appeared disordered arrangement of cells.In DM group, different stages of apoptosis were observed by transmission electron microscope and got worse gradually with its extension.In LOM group, compared with DM group in the same period, RGCs apoptosis signs diminished gradually at 8th and 12th wk.TUNEL detection: no apoptotic RGCs was observed in CON group.In DM group, few TUNEL positive RGCs were seen at 4 th wk, and became more and more gradually. The apoptosis index was significantly higher in DM group compared with CON group in same time and there was statistical significance (P<0.01).In LOM group at 8th and 12th wk, compared with DM group in the same period, the numbers of TUNEL positive RGCs decreased.The apoptosis index was significantly lower in LOM group compared with DM group in the same period (P<0.01). Calcium ion concentration detection by laser confocal microscope: compared with CON group in same time points, at 8th and 12th week, DM group's calcium fluorescent staining intensity of RGCs markedly elevated and had significant differences.( P <0.01 ). In LOM group, at 8th and 12th wk, calcium fluorescent staining intensity of RGCs markedly decreased and had statistical significance(P<0.01). CONCLUSION: The LOM played a protective role for RGCs in early stage of diabetic rats.