泸州医学院学报
瀘州醫學院學報
로주의학원학보
JOURNAL OF LUZHOU MEDICAL COLLEGE
2015年
1期
7-10
,共4页
哮喘%沉默转导子与转录活化子1%RNA干扰
哮喘%沉默轉導子與轉錄活化子1%RNA榦擾
효천%침묵전도자여전록활화자1%RNA간우
Asthma%Signal transducer and activator of transcription 1%RNA interference
目的：：探讨RNA干扰(RNA interference, RNAi)技术沉默转导子与转录活化子1(signal transducer and activator of transcription 1, STAT1)基因对支气管哮喘小鼠气道炎症的影响。方法：将32只4周龄BALB/c雌鼠随机分为正常组、PBS组、空质粒(随机片段RNAi)组和siRNA (pGSTAT1-2)组,利用RNAi沉默支气管哮喘小鼠中STAT1基因,采用免疫组织化学检测STAT1蛋白的表达；通过单肺灌洗留取肺泡灌洗液(BALF),应用酶联免疫吸附试验法(ELISA)检测气道炎症介质如γ干扰素(IFN-γ)和白细胞介素5(IL-5)的表达水平。结果：siRNA组的IFN-γ浓度高于PBS组、空质粒组(P<0.05),而IL-5的表达水平低于PBS组、空质粒组(P<0.05),二者在PBS组和空质粒组组间均无差异(P跃0.05)；siRNA组BALF中嗜酸性粒细胞(EOS)数量和IL-5浓度较空质粒组和PBS组中均明显减低,而IFN-γ的浓度增高,IFN-γ与EOS呈负相关,IL-5与EOS呈正相关,siRNA组BALF中白细胞(WBC)数量较空质粒组和PBS组中均明显减低；STAT-1蛋白主要表达于气道上皮细胞,siRNA组STAT-1蛋白表达明显低于PBS组与空质粒组,正常组低于siRNA组、PBS组和空质粒组,有统计学意义(P<0.05)。结论：siRNA能有效抑制哮喘小鼠气道上皮细胞STAT1蛋白的表达,减轻哮喘小鼠的气道炎症反应。
目的：：探討RNA榦擾(RNA interference, RNAi)技術沉默轉導子與轉錄活化子1(signal transducer and activator of transcription 1, STAT1)基因對支氣管哮喘小鼠氣道炎癥的影響。方法：將32隻4週齡BALB/c雌鼠隨機分為正常組、PBS組、空質粒(隨機片段RNAi)組和siRNA (pGSTAT1-2)組,利用RNAi沉默支氣管哮喘小鼠中STAT1基因,採用免疫組織化學檢測STAT1蛋白的錶達；通過單肺灌洗留取肺泡灌洗液(BALF),應用酶聯免疫吸附試驗法(ELISA)檢測氣道炎癥介質如γ榦擾素(IFN-γ)和白細胞介素5(IL-5)的錶達水平。結果：siRNA組的IFN-γ濃度高于PBS組、空質粒組(P<0.05),而IL-5的錶達水平低于PBS組、空質粒組(P<0.05),二者在PBS組和空質粒組組間均無差異(P躍0.05)；siRNA組BALF中嗜痠性粒細胞(EOS)數量和IL-5濃度較空質粒組和PBS組中均明顯減低,而IFN-γ的濃度增高,IFN-γ與EOS呈負相關,IL-5與EOS呈正相關,siRNA組BALF中白細胞(WBC)數量較空質粒組和PBS組中均明顯減低；STAT-1蛋白主要錶達于氣道上皮細胞,siRNA組STAT-1蛋白錶達明顯低于PBS組與空質粒組,正常組低于siRNA組、PBS組和空質粒組,有統計學意義(P<0.05)。結論：siRNA能有效抑製哮喘小鼠氣道上皮細胞STAT1蛋白的錶達,減輕哮喘小鼠的氣道炎癥反應。
목적：：탐토RNA간우(RNA interference, RNAi)기술침묵전도자여전록활화자1(signal transducer and activator of transcription 1, STAT1)기인대지기관효천소서기도염증적영향。방법：장32지4주령BALB/c자서수궤분위정상조、PBS조、공질립(수궤편단RNAi)조화siRNA (pGSTAT1-2)조,이용RNAi침묵지기관효천소서중STAT1기인,채용면역조직화학검측STAT1단백적표체；통과단폐관세류취폐포관세액(BALF),응용매련면역흡부시험법(ELISA)검측기도염증개질여γ간우소(IFN-γ)화백세포개소5(IL-5)적표체수평。결과：siRNA조적IFN-γ농도고우PBS조、공질립조(P<0.05),이IL-5적표체수평저우PBS조、공질립조(P<0.05),이자재PBS조화공질립조조간균무차이(P약0.05)；siRNA조BALF중기산성립세포(EOS)수량화IL-5농도교공질립조화PBS조중균명현감저,이IFN-γ적농도증고,IFN-γ여EOS정부상관,IL-5여EOS정정상관,siRNA조BALF중백세포(WBC)수량교공질립조화PBS조중균명현감저；STAT-1단백주요표체우기도상피세포,siRNA조STAT-1단백표체명현저우PBS조여공질립조,정상조저우siRNA조、PBS조화공질립조,유통계학의의(P<0.05)。결론：siRNA능유효억제효천소서기도상피세포STAT1단백적표체,감경효천소서적기도염증반응。
Objective: To discuss the effects of RNA interference (RNAi) silencing the gene of signal transducer and activator of transcription 1(STAT1) on airway inflammation of bronchial asthmatic mice. Methods:4 weeks old female BALB/c from 32 mice were randomly divided into normal group, PBS group, empty plasmid ( random fragment RNAi ) group and siRNA (pGSTAT1-2) group. RNAi was used to silence the gene of STAT1 of bronchial asthmatic mice. S-P method was used to detect the expression of STAT1 and ELISA was used to detect the concentration of airway inflammation medium such as IFN-γ and IL-5 from the bronchoalveolar lavage fluid (BALF) by the single lung lavage. Results: The concentration of IFN-γ of siRNA group was higher than that of PBS group and empty plasmid group (P<0.05), but the concentration of IL-5 was opposite (P<0.05). There was no significant difference between PBS group and empty plasmid group (P 跃 0.05). The count of EOS and the concentration of IL-5 were significantly decreased in empty plasmid group and the PBS group, but the concentration of IFN-γ increased, indicating negative correlation between IFN-γ and EOS, and positively correlation between IL-5 and EOS. The count of WBC in BALF of siRNA group was lower than that of PBS group and empty plasmid group (P< 0.05). STAT-1 was expressed in airway epithelial cells, the expression of STAT-1 of siRNA group was significantly lower than that of PBS group and empty plasmid group, while that of the normal group was the lowest (P< 0.05). Conclusion: SiRNA could effectively inhibit the expression of STAT1 of airway epithelial cell and alleviate airway inflammation in asthmatic mice.
