北华大学学报（自然科学版）
北華大學學報（自然科學版）
북화대학학보（자연과학판）
JOURNAL OF BEIHUA UNIVERSITY(NATURAL SCIENCE)
2015年
1期
38-42
,共5页
基因沉默%白细胞介素-23%支气管哮喘%卵蛋白%支气管肺泡灌洗液
基因沉默%白細胞介素-23%支氣管哮喘%卵蛋白%支氣管肺泡灌洗液
기인침묵%백세포개소-23%지기관효천%란단백%지기관폐포관세액
gene silencing%interleukin 23%bronchial asthma%ovalbumin protein%bronchoalveolar lavage
目的：观察白细胞介素23(IL-23)基因沉默对肺组织IL-23蛋白表达、哮喘鼠气道炎症和支气管肺泡灌洗液( BALF)中干扰素( IFN-γ)、白细胞介素-5( IL-5)、肿瘤坏死因子-α( TNF-α)、细胞间黏附分子-1( ICAM-1)水平的影响.方法构建siRNA IL-23表达载体并通过脂质体转染获得高浓度的含有siRNA IL-23的浓缩液,RT-PCR和WB法检测IL-23mRNA及IL-23蛋白表达水平,确定转染成功.通过滴鼻方式吸入到卵蛋白( OVA)致敏的小鼠体内,观察其对BALF中炎症细胞的影响；ELISA方法检测支气管肺泡灌洗液( BALF)中细胞因子IFN-γ,IL-5, TNF-α,ICAM-1的变化；HE染色观察肺组织病理学改变；WB法检测肺组织中IL-23蛋白表达.结果与对照组相比,模型组及空质粒组BALF中IL-5,TNF-α,ICAM-1水平升高,IFN-γ水平下降,肺组织中IL-23蛋白表达量升高(P<0.01),沉默组上述各项指标明显下降,IFN-γ水平升高(P<0.01)；病理学检测结果表明：与对照组相比,模型组和空质粒组气道内有大量炎性细胞浸润,沉默组炎症受到抑制(P<0.01).结论 IL-23在哮喘发病中起重要作用,阻断其基因表达可以明显改变哮喘症状.
目的：觀察白細胞介素23(IL-23)基因沉默對肺組織IL-23蛋白錶達、哮喘鼠氣道炎癥和支氣管肺泡灌洗液( BALF)中榦擾素( IFN-γ)、白細胞介素-5( IL-5)、腫瘤壞死因子-α( TNF-α)、細胞間黏附分子-1( ICAM-1)水平的影響.方法構建siRNA IL-23錶達載體併通過脂質體轉染穫得高濃度的含有siRNA IL-23的濃縮液,RT-PCR和WB法檢測IL-23mRNA及IL-23蛋白錶達水平,確定轉染成功.通過滴鼻方式吸入到卵蛋白( OVA)緻敏的小鼠體內,觀察其對BALF中炎癥細胞的影響；ELISA方法檢測支氣管肺泡灌洗液( BALF)中細胞因子IFN-γ,IL-5, TNF-α,ICAM-1的變化；HE染色觀察肺組織病理學改變；WB法檢測肺組織中IL-23蛋白錶達.結果與對照組相比,模型組及空質粒組BALF中IL-5,TNF-α,ICAM-1水平升高,IFN-γ水平下降,肺組織中IL-23蛋白錶達量升高(P<0.01),沉默組上述各項指標明顯下降,IFN-γ水平升高(P<0.01)；病理學檢測結果錶明：與對照組相比,模型組和空質粒組氣道內有大量炎性細胞浸潤,沉默組炎癥受到抑製(P<0.01).結論 IL-23在哮喘髮病中起重要作用,阻斷其基因錶達可以明顯改變哮喘癥狀.
목적：관찰백세포개소23(IL-23)기인침묵대폐조직IL-23단백표체、효천서기도염증화지기관폐포관세액( BALF)중간우소( IFN-γ)、백세포개소-5( IL-5)、종류배사인자-α( TNF-α)、세포간점부분자-1( ICAM-1)수평적영향.방법구건siRNA IL-23표체재체병통과지질체전염획득고농도적함유siRNA IL-23적농축액,RT-PCR화WB법검측IL-23mRNA급IL-23단백표체수평,학정전염성공.통과적비방식흡입도란단백( OVA)치민적소서체내,관찰기대BALF중염증세포적영향；ELISA방법검측지기관폐포관세액( BALF)중세포인자IFN-γ,IL-5, TNF-α,ICAM-1적변화；HE염색관찰폐조직병이학개변；WB법검측폐조직중IL-23단백표체.결과여대조조상비,모형조급공질립조BALF중IL-5,TNF-α,ICAM-1수평승고,IFN-γ수평하강,폐조직중IL-23단백표체량승고(P<0.01),침묵조상술각항지표명현하강,IFN-γ수평승고(P<0.01)；병이학검측결과표명：여대조조상비,모형조화공질립조기도내유대량염성세포침윤,침묵조염증수도억제(P<0.01).결론 IL-23재효천발병중기중요작용,조단기기인표체가이명현개변효천증상.
Objective To observe the effects of gene silencing of interleukin 23 ( IL-23 ) on IL-23 protein expression in lung tissue,airway inflammation in asthmatic rat and the levels of interferon-γ( IFN-γ) ,interleukin 5(IL-5),tumor necrosis factor alpha(TNF-α),intercellular adhesion molecule-1(ICAM-1)in bronchoalveolar lavage fluid( BALF) . Methods siRNA IL-23 expression vector was constructed and high concentration of siRNA containing IL- 23 concentrate was gained by liposome transfection technology;RT-PCR and WB method were used to test the levels of IL-23 mRNA and IL-23 protein expression in order to identify the successful transfections. Mice was sensitized by intranasal ovalbumin( OVA) inhalation to observe its effects on inflammatory cells in BALF. Changes of IFN-γ, IL-5, TNF-α and ICAM-1 were detected via ELISA method. Pathological change was observed by HE staining and IL-23 protein expression was detected by WB in lung tissue. Results Compared with those in the control group,IL-5,TNF-α,ICAM-1 levels of the model group and the empty plasmid group increased,IFN-γ level declined in BALF and IL-23 protein expression in lung tissue rised(P<0. 01). The above-mentioned indicators in silent group significantly decreased while IFN-γ level was enhanced ( P<0 . 01 ) . Pathological results showed that compared with the control group, model group and the empty plasmid group airway contained a large number of inflammatory cells infiltration,while the inflammation response in silent group was inhibited(P<0. 01). Conclusion IL-23 may play an important role in the pathogenesis of asthma,blocking its gene expression could obviously relief the asthmatic symptoms.
