天津医药
天津醫藥
천진의약
TIANJIN MEDICAL JOURNAL
2014年
3期
217-219
,共3页
糖尿病%视网膜神经节细胞%RNA,小分子干扰%细胞凋亡%丙二醛%半胱氨酸天冬氨酸蛋白酶3%疾病模型,动物%Nogo受体
糖尿病%視網膜神經節細胞%RNA,小分子榦擾%細胞凋亡%丙二醛%半胱氨痠天鼕氨痠蛋白酶3%疾病模型,動物%Nogo受體
당뇨병%시망막신경절세포%RNA,소분자간우%세포조망%병이철%반광안산천동안산단백매3%질병모형,동물%Nogo수체
diabetes mellitus%retinal ganglion cells%RNA,small interfering%apoptosis%malondialdehyde%caspase 3%disease models,animal%Nogo receptor
目的：探讨Nogo受体对糖尿病大鼠视网膜神经节细胞（RGC）凋亡的影响及机制。方法 SD大鼠腹腔注射链脲佐菌素诱导30只糖尿病模型。分为对照组、糖尿病组、siNgR组、siRNA空白组，每组10只。siNgR组糖尿病大鼠玻璃体内注射Nogo受体反义核苷酸序列；siRNA空白组糖尿病大鼠玻璃体内注射阴性核苷酸序列。1个月后，免疫荧光组化检测Nogo受体与RGC标志物Brn3a的共存；以TUNEL染色观察RGC凋亡；试剂盒检测视网膜丙二醛（MDA）含量；Western blot检测视网膜Nogo受体及半胱氨酸天冬氨酸蛋白酶3（caspase-3）表达。结果 Nogo受体大量表达于视网膜RGC内，对照组、糖尿病组、siRNA空白组及siNgR组MDA含量分别为（3.68±0.47）、（8.07±1.24）、（7.54±1.53）及（5.12±0.62）μmol/g蛋白，RGC凋亡率分别为（5.1±0.2）%、（49.3±2.7）%、（45.6±1.8）%及（12.4±0.6）%， Nogo受体相对表达量分别为（0.18±0.07）%、（0.45±0.12）%、（0.40±0.09）%及（0.16±0.09）%，caspase-3相对表达量分别为（0.16±0.05）%、（0.40±0.18）%、（0.42±0.12）%及（0.17±0.08）%。与对照组相比，糖尿病组及siRNA空白组视网膜RGC凋亡明显，Nogo受体及caspase-3表达上调，MDA含量增加（P<0.05）。与糖尿病组相比siNgR组视网膜RGC凋亡减少、Nogo受体及caspase-3表达下调、MDA含量降低（P<0.05）。结论 Nogo受体表达上调参与了糖尿病视网膜RGC凋亡，其机制可能与Nogo受体上调caspase-3表达、诱发视网膜氧化应激有关。
目的：探討Nogo受體對糖尿病大鼠視網膜神經節細胞（RGC）凋亡的影響及機製。方法 SD大鼠腹腔註射鏈脲佐菌素誘導30隻糖尿病模型。分為對照組、糖尿病組、siNgR組、siRNA空白組，每組10隻。siNgR組糖尿病大鼠玻璃體內註射Nogo受體反義覈苷痠序列；siRNA空白組糖尿病大鼠玻璃體內註射陰性覈苷痠序列。1箇月後，免疫熒光組化檢測Nogo受體與RGC標誌物Brn3a的共存；以TUNEL染色觀察RGC凋亡；試劑盒檢測視網膜丙二醛（MDA）含量；Western blot檢測視網膜Nogo受體及半胱氨痠天鼕氨痠蛋白酶3（caspase-3）錶達。結果 Nogo受體大量錶達于視網膜RGC內，對照組、糖尿病組、siRNA空白組及siNgR組MDA含量分彆為（3.68±0.47）、（8.07±1.24）、（7.54±1.53）及（5.12±0.62）μmol/g蛋白，RGC凋亡率分彆為（5.1±0.2）%、（49.3±2.7）%、（45.6±1.8）%及（12.4±0.6）%， Nogo受體相對錶達量分彆為（0.18±0.07）%、（0.45±0.12）%、（0.40±0.09）%及（0.16±0.09）%，caspase-3相對錶達量分彆為（0.16±0.05）%、（0.40±0.18）%、（0.42±0.12）%及（0.17±0.08）%。與對照組相比，糖尿病組及siRNA空白組視網膜RGC凋亡明顯，Nogo受體及caspase-3錶達上調，MDA含量增加（P<0.05）。與糖尿病組相比siNgR組視網膜RGC凋亡減少、Nogo受體及caspase-3錶達下調、MDA含量降低（P<0.05）。結論 Nogo受體錶達上調參與瞭糖尿病視網膜RGC凋亡，其機製可能與Nogo受體上調caspase-3錶達、誘髮視網膜氧化應激有關。
목적：탐토Nogo수체대당뇨병대서시망막신경절세포（RGC）조망적영향급궤제。방법 SD대서복강주사련뇨좌균소유도30지당뇨병모형。분위대조조、당뇨병조、siNgR조、siRNA공백조，매조10지。siNgR조당뇨병대서파리체내주사Nogo수체반의핵감산서렬；siRNA공백조당뇨병대서파리체내주사음성핵감산서렬。1개월후，면역형광조화검측Nogo수체여RGC표지물Brn3a적공존；이TUNEL염색관찰RGC조망；시제합검측시망막병이철（MDA）함량；Western blot검측시망막Nogo수체급반광안산천동안산단백매3（caspase-3）표체。결과 Nogo수체대량표체우시망막RGC내，대조조、당뇨병조、siRNA공백조급siNgR조MDA함량분별위（3.68±0.47）、（8.07±1.24）、（7.54±1.53）급（5.12±0.62）μmol/g단백，RGC조망솔분별위（5.1±0.2）%、（49.3±2.7）%、（45.6±1.8）%급（12.4±0.6）%， Nogo수체상대표체량분별위（0.18±0.07）%、（0.45±0.12）%、（0.40±0.09）%급（0.16±0.09）%，caspase-3상대표체량분별위（0.16±0.05）%、（0.40±0.18）%、（0.42±0.12）%급（0.17±0.08）%。여대조조상비，당뇨병조급siRNA공백조시망막RGC조망명현，Nogo수체급caspase-3표체상조，MDA함량증가（P<0.05）。여당뇨병조상비siNgR조시망막RGC조망감소、Nogo수체급caspase-3표체하조、MDA함량강저（P<0.05）。결론 Nogo수체표체상조삼여료당뇨병시망막RGC조망，기궤제가능여Nogo수체상조caspase-3표체、유발시망막양화응격유관。
Objective To investigate the effect of Nogo receptor on the apoptosis of retinal ganglion cell (RGC) in di-abetic rats, and the potential mechanism thereof. Methods Thirty diabetic model rats were induced by intraperitoneal ad-ministration of streptozotocin. Model rats were randomly divided into control group, diabetes mellitus (DM) group, siNgR group and siRNA control group (n=10 for each group). Diabetic rats in siNgR group were intravitreally administrated with No-go receptor antisense nucleotide. Diabetic rats in siRNA control group were intravitreally administrated with negative nucleo-tide. One month after diabetes onset, colocalization of Nogo receptor and Brn3a (marker of RGC) was observed by immunohis-tochemistry. The apoptosis of RGC was detected by TUNEL staining. The level of retinal malondialdehyde (MDA) was ob-served with kit, and the expressions of Nogo receptor and caspase-3 were detected with Western blot assay. Results It was found that the Nogo receptor was highly expressed in RGC. The levels of retinal MDA were (3.68±0.47), (8.07±1.24), (7.54± 1.53) and (5.12 ± 0.62) μmol/g protein for control group, DM group, siRNA control group and siNgR group. The apoptotic rates of RGC were (5.1 ± 0.2)%, (49.3 ± 2.7)%, (45.6 ± 1.8)%and (12.4 ± 0.6)%respectively. The expressions of Nogo receptor were (0.18 ± 0.07)%, (0.45 ± 0.12)%, (0.40 ± 0.09)%and (0.16 ± 0.09)%. The expressions of caspase-3 were (0.16 ± 0.05)%, (0.40±0.18)%, (0.42±0.12)%and (0.17±0.08)%. Compared with control group, there was significant increase in apoptosis of RGC, significantly up-regulated expressions in Nogo receptor and caspase-3, and significantly increased level of MDA in DM group and siRNA control group(P<0.05). Compared with DM group, there were decreased apoptotic rate of RGC, de-creased expressions of Nogo receptor and caspase-3, and decreased level of retinal MDA in siNgR group (P<0.05). Conclu-sion The increased level of Nogo receptor induces oxidative stress and up-regulation of caspase-3 in diabetic retina, play-ing an important role in the apoptosis of RGC.
