CAJ | 학술논문

目的研究IL-10对脂多糖(LPS)诱导的Hela细胞IL-15mRNA和IL-6mRNA表达的影响,分析其激活的信号转导通路.方法培养的Hela细胞,经不同浓度的LPS及IL-10单独或联合处理后,提取细胞总RNA和总蛋白,RT-PCR分析IL-15和IL-6转录水平的变化,Western blot分析信号转导通路蛋白变化.结果 RT-PCR分析得知①1 ng～10 μg LPS刺激Hela细胞12 h后,IL-15mRNA和IL-6mRNA水平均明显上调(与对照组比较:P＜0.01),且存在剂量依赖关系,100 ng/mL时达峰值;100 ng/mL LPS刺激Hela细胞0～24 h,IL-15mRNA和IL-6mRNA水平亦明显上调(与对照组比较:P＜0.01),24 h内存在时间依赖关系,12 h时达峰值.②单独IL-10(10 ng/mL)作用于Heh细胞12 h后,IL-15mRNA和IL-6mRNA没有明显变化(与对照组比较:P＞0.05).不同浓度的IL-10(1,10,100 ng/mL)均下调100 ng/mL LPS诱导的Hela细胞IL-15mRNA和IL-6mRNA的表达,且浓度越高IL-10抑制作用越明显.Western blot 分析显示LPS主要通过磷酸化信号蛋白PI3K/AKT和ERK1/2上调IL-15和IL-6转录,IL-10能阻断AKT的磷酸化而对ERK1/2的磷酸化没有影响.结论 IL-10可抑制LPS诱导的炎症细胞因子IL-15和IL-6的转录,这可能与其阻断AKT的磷酸化有关,因而,IL-10可能应用于某些临床感染性疾病的预防和治疗.
목적 연구IL-10대지다당(LPS)유도적Hela세포IL-15mRNA화IL-6mRNA표체적영향,분석기격활적신호전도통로.방법 배양적Hela세포,경불동농도적LPS급IL-10단독혹연합처리후,제취세포총RNA화총단백,RT-PCR분석IL-15화IL-6전록수평적변화,Western blot분석신호전도통로단백변화.결과 RT-PCR분석득지①1 ng～10 μg LPS자격Hela세포12 h후,IL-15mRNA화IL-6mRNA수평균명현상조(여대조조비교:P＜0.01),차존재제량의뢰관계,100 ng/mL시체봉치;100 ng/mL LPS자격Hela세포0～24 h,IL-15mRNA화IL-6mRNA수평역명현상조(여대조조비교:P＜0.01),24 h내존재시간의뢰관계,12 h시체봉치.②단독IL-10(10 ng/mL)작용우Heh세포12 h후,IL-15mRNA화IL-6mRNA몰유명현변화(여대조조비교:P＞0.05).불동농도적IL-10(1,10,100 ng/mL)균하조100 ng/mL LPS유도적Hela세포IL-15mRNA화IL-6mRNA적표체,차농도월고IL-10억제작용월명현.Western blot 분석현시LPS주요통과린산화신호단백PI3K/AKT화ERK1/2상조IL-15화IL-6전록,IL-10능조단AKT적린산화이대ERK1/2적린산화몰유영향.결론 IL-10가억제LPS유도적염증세포인자IL-15화IL-6적전록,저가능여기조단AKT적린산화유관,인이,IL-10가능응용우모사림상감염성질병적예방화치료.
Purpose To investigate the effect of interleukin-10(IL-10)on IL-15mRNA and IL-6mRNA in Hela cells induced by lipopolysaccharide(LPS)and to analyze their activiated signal transduction pathways.Methods Extracted total RNA and total proteins of cultured Hela cells,which were treated with different concentrations of LPS,or IL-10 alone or in combined use,were to analyze the levels of IL-15mRNA and IL-6mRNA using RT-PCR and to analyze the expression of signal transduction pathway proteins using Western blot.Results RT-PCR indicated that expression of IL-15mRNA and IL-6mRNA in Hela cells strikingly increased after 12 h using 1 ng-10μg LPS(P＜0.01 vs control),which had dose-dependence and achieved peak value using 100 ng/mL.During 0-24 h,expression of IL-15mRNA and IL-6mRNA strikingly increased with time changing(P＜0.01 vs control),which had time-dependence and achieved peak value at 12 h.Expression of IL-15mRNA and IL-6mRNA had no conspicuous difference in Hela cells treated with IL-10(10 ng/mL)alone(P＞0.05 vs control).Different concentrations of IL-10 1,10,100 ns/mL)markedly downregnlated expression of IL-15mRNA and IL-6mRNA in Hela cells induced by LPS.Furthermore,the effect of inhibition will be more obvious with dose increasing.Western blot indicated that LPS upregulated expression of IL-15mRNA and IL-6mRNA by phosphorylation of PI3K/AKT and ERK1/2.IL-10 blocked phosphorylation of AKT,but could not affect phosphorylation of ERK1/2.Conclusion IL-10 downregulated the expression of inflammatory cytokines IL-15 and 1L-6 induced by LPS,which may correlate with the fact that phosphorylation of AKT was blcoked by IL-10.Therefore,IL-10 may be used to prevent and treat some clincal infective diseases.