中华创伤杂志
中華創傷雜誌
중화창상잡지
Chinese Journal of Traumatology
2014年
6期
616-620
,共5页
宋平%袁容娣%陈春林%韩非%孙敏%叶剑
宋平%袁容娣%陳春林%韓非%孫敏%葉劍
송평%원용제%진춘림%한비%손민%협검
视神经%视网膜神经节细胞%免疫球蛋白类
視神經%視網膜神經節細胞%免疫毬蛋白類
시신경%시망막신경절세포%면역구단백류
Optic nerve%Retinal ganglion cells%Immunoglobulins
目的 研究配对免疫球蛋白样受体B(paired immunoglobulin-like receptor B,PirB)对体外视网膜神经节细胞(retinal ganglion cells,RGCs)生长的影响. 方法 分离野生型C57B J/6小鼠的RGCs,免疫荧光检测RGCs中PirB的表达情况,荧光定量PCR和Western blot方法检测培养1,3,5,7,9d时PirB在RGCs中的表达水平,并将培养的原代RGCs分为对照组(对RGCs不加处理)、慢病毒转染干扰PirB组(A组)、慢病毒转染干扰NgR组(B组)、慢病毒转染干扰PirB且联用睫状神经营养因子(ciliary neurotrophic factor,CNTF)组(C组)、慢病毒转染干扰NgR且联用CNTF组(D组)和仅CNTF处理组(E组).使用MTT法及从形态学方面观察RGCs的生长发育情况. 结果 免疫荧光确定PirB在RGCs中表达,其mRNA和蛋白水平随时间呈现先升高后下降的变化,分别于培养5d和7d时达最高水平.A组和B组PirB或NgR的表达降低,A组RGCs的生长状态较对照组旺盛:3d时为(40.2±5.3)μm:(29.1±3.8)μm,5d时为(72.2±4.2)μm:(52.3±8.2)μm(P ＜0.05);B组、D组和E组与对照组比较、C组与A组比较,RGCs的生长状态差异均无统计学意义(P均＞0.05). 结论 阻断PirB可明显改善RGCs的生长状态,为临床治疗视神经损伤提供了新的思路.
目的 研究配對免疫毬蛋白樣受體B(paired immunoglobulin-like receptor B,PirB)對體外視網膜神經節細胞(retinal ganglion cells,RGCs)生長的影響. 方法 分離野生型C57B J/6小鼠的RGCs,免疫熒光檢測RGCs中PirB的錶達情況,熒光定量PCR和Western blot方法檢測培養1,3,5,7,9d時PirB在RGCs中的錶達水平,併將培養的原代RGCs分為對照組(對RGCs不加處理)、慢病毒轉染榦擾PirB組(A組)、慢病毒轉染榦擾NgR組(B組)、慢病毒轉染榦擾PirB且聯用睫狀神經營養因子(ciliary neurotrophic factor,CNTF)組(C組)、慢病毒轉染榦擾NgR且聯用CNTF組(D組)和僅CNTF處理組(E組).使用MTT法及從形態學方麵觀察RGCs的生長髮育情況. 結果 免疫熒光確定PirB在RGCs中錶達,其mRNA和蛋白水平隨時間呈現先升高後下降的變化,分彆于培養5d和7d時達最高水平.A組和B組PirB或NgR的錶達降低,A組RGCs的生長狀態較對照組旺盛:3d時為(40.2±5.3)μm:(29.1±3.8)μm,5d時為(72.2±4.2)μm:(52.3±8.2)μm(P ＜0.05);B組、D組和E組與對照組比較、C組與A組比較,RGCs的生長狀態差異均無統計學意義(P均＞0.05). 結論 阻斷PirB可明顯改善RGCs的生長狀態,為臨床治療視神經損傷提供瞭新的思路.
목적 연구배대면역구단백양수체B(paired immunoglobulin-like receptor B,PirB)대체외시망막신경절세포(retinal ganglion cells,RGCs)생장적영향. 방법 분리야생형C57B J/6소서적RGCs,면역형광검측RGCs중PirB적표체정황,형광정량PCR화Western blot방법검측배양1,3,5,7,9d시PirB재RGCs중적표체수평,병장배양적원대RGCs분위대조조(대RGCs불가처리)、만병독전염간우PirB조(A조)、만병독전염간우NgR조(B조)、만병독전염간우PirB차련용첩상신경영양인자(ciliary neurotrophic factor,CNTF)조(C조)、만병독전염간우NgR차련용CNTF조(D조)화부CNTF처리조(E조).사용MTT법급종형태학방면관찰RGCs적생장발육정황. 결과 면역형광학정PirB재RGCs중표체,기mRNA화단백수평수시간정현선승고후하강적변화,분별우배양5d화7d시체최고수평.A조화B조PirB혹NgR적표체강저,A조RGCs적생장상태교대조조왕성:3d시위(40.2±5.3)μm:(29.1±3.8)μm,5d시위(72.2±4.2)μm:(52.3±8.2)μm(P ＜0.05);B조、D조화E조여대조조비교、C조여A조비교,RGCs적생장상태차이균무통계학의의(P균＞0.05). 결론 조단PirB가명현개선RGCs적생장상태,위림상치료시신경손상제공료신적사로.
Objective To determine the effect of paired immunoglobulin-like receptor B (PirB) on growth of retinal ganglion cells (RGCs) in vitro.Methods Expression of PirB in RGCs isolated from wild-type C57 BJ/6 mice was measured by immunofluorescence.Real-time PCR and Western blot were performed to detect PirB expressions in RGCs cultured for 1,3,5,7 and 9 days.Primary cultured RGCs were left untreated as controls,transfected with lentiviral-delivered PirB RNAi as Group A,lentiviral-delivered NgR RNAi as Group B,lentiviral-delivered PirB RNAi plus ciliary neurotrophic factor (CNTF) as Group C,lentiviral-delivered NgR plus CNTF as Group D,and only CNTF as Group E.Growth and development of RGCs were evaluated by MTT assay and morphological analysis.Results Immunofluorescence confirmed expression of PirB in RGCs,with its mRNA and protein levels changing from increase at first to decrease later with time,up to peak at days 5 and 7.PirB and NgR were respectively down-regulated in Groups A and B.Viability of RGCs was improved in Group A compared with control group [(40.2 ± 5.3) μm vs (29.1 ± 3.8) μm at day 3 ; (72.2 ± 4.2) μm vs (52.3 ± 8.2) μm at day 5,both P ＜ 0.05].By contrast,no significant difference was found in viability of RGCs among Groups B,D and E when compared to control group (P ＞ 0.05).Growth of RGCs between Groups A and C revealed insignificant difference (P ＞ 0.05).Conclusion Knockdown of PirB is beneficial for the growth of RGCs,suggesting a novel method to treat optic nerve injury.
