中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
4期
784-786
,共3页
何筱天%吴多光%姜明%王稳健%王铭辉%张惠忠
何篠天%吳多光%薑明%王穩健%王銘輝%張惠忠
하소천%오다광%강명%왕은건%왕명휘%장혜충
闭塞性细支气管炎%气管移植%核因子-κB%小干扰RNA
閉塞性細支氣管炎%氣管移植%覈因子-κB%小榦擾RNA
폐새성세지기관염%기관이식%핵인자-κB%소간우RNA
Bronchiolitis obliterans%Trachea transplantation%Nuclear factor-κB%Small interfering RNA
目的 观察抑制气管移植物核因子-κB (NF-κB)蛋白表达对同种异体气管移植急性排斥反应的影响.方法 取50只BALB/c和C57BL/6小鼠分为3组:同基因移植组(A组)、脂质体对照组(B组)和小干扰RNA (siRNA)干扰组(C组).体外建立siRNA干扰NF-κB蛋白低表达支气管移植物以及气管移植动物模型,最后比较各组移植后第7、14和28天排斥反应的程度.反转录-聚合酶链反应(RT-PCR)和Western blot检测NF-κB基因和蛋白的表达水平,苏木素-伊红(HE)染色检测炎性细胞浸润、气管通畅度和上皮细胞存活率.结果 与A组比较,气管移植后第14天,B组支气管大量炎性细胞浸润,管腔出现纤维阻塞,通畅度为(62.0±16.7)％,支气管上皮细胞存活率为(11.0±24.6)％;而C组少量炎性细胞浸润,支气管管腔通畅度为100％ (P ＜0.05),C组支气管上皮细胞存活率为(78.80±4.41)％(P＜0.05);气管移植后第21天,B、C两组管腔通畅度及上皮细胞存活率均为0.结论 siRNA沉默支气管上皮细胞NF-κB蛋白表达可通过抑制急性排斥反应的水平,延长气管管腔的通畅时间和上皮细胞存活时间,从而延缓闭塞性细支气管炎的发展.
目的 觀察抑製氣管移植物覈因子-κB (NF-κB)蛋白錶達對同種異體氣管移植急性排斥反應的影響.方法 取50隻BALB/c和C57BL/6小鼠分為3組:同基因移植組(A組)、脂質體對照組(B組)和小榦擾RNA (siRNA)榦擾組(C組).體外建立siRNA榦擾NF-κB蛋白低錶達支氣管移植物以及氣管移植動物模型,最後比較各組移植後第7、14和28天排斥反應的程度.反轉錄-聚閤酶鏈反應(RT-PCR)和Western blot檢測NF-κB基因和蛋白的錶達水平,囌木素-伊紅(HE)染色檢測炎性細胞浸潤、氣管通暢度和上皮細胞存活率.結果 與A組比較,氣管移植後第14天,B組支氣管大量炎性細胞浸潤,管腔齣現纖維阻塞,通暢度為(62.0±16.7)％,支氣管上皮細胞存活率為(11.0±24.6)％;而C組少量炎性細胞浸潤,支氣管管腔通暢度為100％ (P ＜0.05),C組支氣管上皮細胞存活率為(78.80±4.41)％(P＜0.05);氣管移植後第21天,B、C兩組管腔通暢度及上皮細胞存活率均為0.結論 siRNA沉默支氣管上皮細胞NF-κB蛋白錶達可通過抑製急性排斥反應的水平,延長氣管管腔的通暢時間和上皮細胞存活時間,從而延緩閉塞性細支氣管炎的髮展.
목적 관찰억제기관이식물핵인자-κB (NF-κB)단백표체대동충이체기관이식급성배척반응적영향.방법 취50지BALB/c화C57BL/6소서분위3조:동기인이식조(A조)、지질체대조조(B조)화소간우RNA (siRNA)간우조(C조).체외건립siRNA간우NF-κB단백저표체지기관이식물이급기관이식동물모형,최후비교각조이식후제7、14화28천배척반응적정도.반전록-취합매련반응(RT-PCR)화Western blot검측NF-κB기인화단백적표체수평,소목소-이홍(HE)염색검측염성세포침윤、기관통창도화상피세포존활솔.결과 여A조비교,기관이식후제14천,B조지기관대량염성세포침윤,관강출현섬유조새,통창도위(62.0±16.7)％,지기관상피세포존활솔위(11.0±24.6)％;이C조소량염성세포침윤,지기관관강통창도위100％ (P ＜0.05),C조지기관상피세포존활솔위(78.80±4.41)％(P＜0.05);기관이식후제21천,B、C량조관강통창도급상피세포존활솔균위0.결론 siRNA침묵지기관상피세포NF-κB단백표체가통과억제급성배척반응적수평,연장기관관강적통창시간화상피세포존활시간,종이연완폐새성세지기관염적발전.
Objective To investigate the effects of nuclear factor-κB (NF-κB) on chronic allograft rejection after trachea transplantation.Methods Fifty BALB/c and C57BL/6 nude mice were divided into 3 groups:transplantation group,lipidosome negative control group,and small interfering RNA (siRNA) group.With the establishment of low-expressed NF-κB transplants,degrees of rejection reaction were compared among these groups at 7,14 and 28 days after transplantation respectively.Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting were used to detect the expression of NF-κB transcriptionally and post-transcriptionally.The tracheal epithelium survival rate and inflammatory cell infiltration rate were quantified by H&E staining.Results At 14th day after trachea transplantation,negative control group showed severe infiltration of inflammatory cells,the patency rate was (62.0 ± 16.7) ％,and survival rate of tracheal epithelia was (11.0 ± 24.6) ％,and siRNA-treated group showed intact trachea epithelium,mild infiltration of inflammatory cells,the patency rate was 100％ (P ＜ 0.05),and survival rate of tracheal epithelia was (78.80 ± 4.41) ％ (P ＜ 0.05).However,at 21 st day after transplantation,both negative control group and siRNA-treated group demonstrated 0％ in both patency rate and survival rate.Conclusion The knock-down of NF-κB expression in the tracheal epithelia can inhibit acute rejection reaction and can alleviate the development of occlusive bronchiolitis.
