CAJ | 학술논문

目的：研究 NgR-Rhoa-Rock 信号通路在糖尿病(diabetes mellitus,DM)大鼠视网膜神经节细胞凋亡中的作用及机制。 　　方法：健康 SD 大鼠,以50mg/ kg 体质量单次腹腔注射1%链脲佐菌素的方法处理,以血糖值大于16.7mmol / L 定为糖尿病模型；然后随机分成3组,每组10只。另取10只健康 SD 大鼠作对照组。再依次对4组大鼠进行双侧眼球玻璃体腔内注射 NgR 小干扰病毒10μL( siRNA 组)、NgR小干扰病毒稀释液10μL(DM 组)、NgR 小干扰病毒阴性对照液10μL( siRNA 空白组)、NgR 小干扰病毒稀释液10μL(正常对照组),正常喂养。期间对血糖、体质量等一些生命指标进行观查、测量并记录。12wk 后,行视网膜Western blot 检测 NgR、Rhoa 的表达,HE 染色、TUNEL 染色。 　　结果：Western 检测：与正常对照组相比,DM 组和 siRNA空白组中 NgR、Rhoa 表达明显增加(P<0.01),而 siRNA组无明显变化(P>0.05)；与 DM 组和 siRNA 空白组相比, siRNA 组 NgR、Rhoa 表达明显下调。 HE 染色：与正常对照组相比,三组模型鼠均有不同程度节细胞排列紊乱、形态不规则、间距不一致；与 DM 组和 siRNA 空白组相比, siRNA 组节细胞在排列顺序、外形大小等有一定程度改善。 TUNEL 染色：与正常对照组相比,三组模型鼠均有节细胞凋亡；与 DM 组和 siRNA 空白组相比,siRNA 组凋亡细胞数量较少,细胞外形有明显改善。 　　结论：DM 时大鼠视网膜 NgR、Rhoa 表达上调,抑制 NgR-Rhoa-Rock 可以改善糖尿病视网膜神经节细胞的凋亡。
목적：연구 NgR-Rhoa-Rock 신호통로재당뇨병(diabetes mellitus,DM)대서시망막신경절세포조망중적작용급궤제。 　　방법：건강 SD 대서,이50mg/ kg 체질량단차복강주사1%련뇨좌균소적방법처리,이혈당치대우16.7mmol / L 정위당뇨병모형；연후수궤분성3조,매조10지。령취10지건강 SD 대서작대조조。재의차대4조대서진행쌍측안구파리체강내주사 NgR 소간우병독10μL( siRNA 조)、NgR소간우병독희석액10μL(DM 조)、NgR 소간우병독음성대조액10μL( siRNA 공백조)、NgR 소간우병독희석액10μL(정상대조조),정상위양。기간대혈당、체질량등일사생명지표진행관사、측량병기록。12wk 후,행시망막Western blot 검측 NgR、Rhoa 적표체,HE 염색、TUNEL 염색。 　　결과：Western 검측：여정상대조조상비,DM 조화 siRNA공백조중 NgR、Rhoa 표체명현증가(P<0.01),이 siRNA조무명현변화(P>0.05)；여 DM 조화 siRNA 공백조상비, siRNA 조 NgR、Rhoa 표체명현하조。 HE 염색：여정상대조조상비,삼조모형서균유불동정도절세포배렬문란、형태불규칙、간거불일치；여 DM 조화 siRNA 공백조상비, siRNA 조절세포재배렬순서、외형대소등유일정정도개선。 TUNEL 염색：여정상대조조상비,삼조모형서균유절세포조망；여 DM 조화 siRNA 공백조상비,siRNA 조조망세포수량교소,세포외형유명현개선。 　　결론：DM 시대서시망막 NgR、Rhoa 표체상조,억제 NgR-Rhoa-Rock 가이개선당뇨병시망막신경절세포적조망。
To study the function and mechanism of the NgR-Rhoa- Rock signal pathways which exists in the retinal ganglion cells apoptosis in diabetes mellitus (DM) rats. ● METHODS: Some healthy SD rats were operated by means of single intraperitoneal injection of 1%streptozotocin based on the standard of 50mg/ kg wight, after that the blood sugar value was greater than 16. 7mmol/ L as DM model, then randomly divided into 3 groups, each group was 10 rats. ln addition to take 10 healthy SD rats as control group. Four groups of rats were bilaterally eyeball intravitreal injection in turn with NgR-siRNA virus 10μ L (siRNA group), NgR-siRNA virus diluted 10μ L ( DM group), NgR - siRNA virus - negative -control solution 10μ L (siRNA blank group), NgR- siRNA virus diluted 10μ L ( normal control group ), and fed normally. During that time, some life indexes like blood glucose, body mass, etc. were measured and recorded. After 12wk, the expression of NgR and Rhoa, HE staining, and TUNNEL staining were detected by Western blot analysis. ● RESULTS: Western blot analysis: compared with normal control group, the expression of NgR and Rhoa in DM group and siRNA blank group increased significantly (P<0. 01), while siRNA group was no significant change (P > 0. 05); compared with DM group and siRNA blank group, the expression of those proteins significantly lowered in siRNA group. HE staining: compared with normal control group, some extent ganglion cells arranged disorder, irregular shape, spacing not consistent were all found in three groups of model rats;compared with DM group and siRNA blank group, there was some improvement in siRNA group of ganglion cells about the order and shape size. TUNEL staining:compared with normal control group, there were retinal ganglion cells apoptosis in all of three groups of model rats. Compared with DM group and siRNA blank group, the number of retinal ganglion cells apoptotic cells was less, and the shape of cells had improved significantly in siRNA group. ●CONCLUSlON: ln the DM phase, the expression of NgR and Rhoa were up - regulation, the condition of diabetic retinal ganglion cell apoptosis was improved after that the NgR-Rhoa-Rock signal pathways had been inhibited.