中华实用儿科临床杂志
中華實用兒科臨床雜誌
중화실용인과림상잡지
Journal of Applied Clinical Pediatrics
2015年
7期
490-493
,共4页
李晶%周伟%袁伟明%黄龙光%郑少伟%唐娟
李晶%週偉%袁偉明%黃龍光%鄭少偉%唐娟
리정%주위%원위명%황룡광%정소위%당연
肠上皮细胞%大鼠%脂多糖%白细胞介素-6%白细胞介素-1β%肿瘤坏死因子-α%细胞增殖
腸上皮細胞%大鼠%脂多糖%白細胞介素-6%白細胞介素-1β%腫瘤壞死因子-α%細胞增殖
장상피세포%대서%지다당%백세포개소-6%백세포개소-1β%종류배사인자-α%세포증식
Intestinal epithelial cell%Rat%Lipopolysaccharide%Interleukin-6%Interleukin-1 β%Tumor necrosis factor-α%Cell proliferation
目的 探讨不同浓度脂多糖(LPS)对大鼠肠上皮细胞(IEC-6)增殖和白细胞介素(IL)-6、IL-1β及肿瘤坏死因子-α(TNF-α)分泌的影响.方法 体外培养的IEC-6分为正常对照组(0 mg/L,A组)、0.1 mg/L组(B组)、0.5 mg/L组(C组)、1.0 mg/L组(D组)、5.0 mg/L组(E组)和10.0 mg/L组(F组),分别用对应浓度的LPS与细胞共培养3h、5h、7h后,噻唑蓝(MTT)比色法检测细胞增殖率,酶联免疫吸附试验(ELISA)检测细胞培养上清中IL-6、IL-1β及TNF-α的质量浓度.结果 LPS与IEC-6共培养3h、5h、7h时,随着LPS浓度的增加,IEC-6的增殖率逐渐降低.共培养3h和5h时,B组、C组、D组、E组和F组增殖率分别与A组比较,差异均无统计学意义(P均>0.05);共培养7h时,B组、C组、D组增殖率与A组比较,差异均无统计学意义(P均>0.05),E组、F组增殖率与A组相比,差异有统计学意义(t=4.216,P=0.014;t=14.991,P=0.000).E组和F组共培养7h时的细胞增殖率较共培养5h时降低,差异均有统计学意义(t=2.576,P=0.033;t2.975,P=0.018);而共培养5h时E组与A组比较,差异无统计学意义(P>0.05).LPS与IEC-6共培养3h、5h和7h时,B组、C组、D组、E组和F组共培养上清中IL-6水平与A组比较均有升高,差异均有统计学意义(P均<0.01),其中各时间点E组IL-6水平均最高.E组LPS与IEC-6共培养5h时IL-6水平出现峰值.LPS与IEC-6共培养3h、5h和7h时,B组、C组、D组、E组、F组共培养上清中IL-1β、TNF-α水平与A组比较,差异均无统计学意义(P均>0.05).结论 在一定浓度、一定培养时间范围内,随LPS浓度的增加,IEC-6增殖率呈下降趋势.0~10.0 mg/L的LPS与IEC-6共培养,可刺激IEC-6分泌IL-6.5.0 mg/L的LPS与IEC-6共培养5h,其培养上清中IL-6水平最高.LPS对IEC-6培养上清中TNF-α和IL-1β水平无影响.5.0 mg/L的LPS与IEC-6共培养5h可制作较好的IEC-6炎性模型.
目的 探討不同濃度脂多糖(LPS)對大鼠腸上皮細胞(IEC-6)增殖和白細胞介素(IL)-6、IL-1β及腫瘤壞死因子-α(TNF-α)分泌的影響.方法 體外培養的IEC-6分為正常對照組(0 mg/L,A組)、0.1 mg/L組(B組)、0.5 mg/L組(C組)、1.0 mg/L組(D組)、5.0 mg/L組(E組)和10.0 mg/L組(F組),分彆用對應濃度的LPS與細胞共培養3h、5h、7h後,噻唑藍(MTT)比色法檢測細胞增殖率,酶聯免疫吸附試驗(ELISA)檢測細胞培養上清中IL-6、IL-1β及TNF-α的質量濃度.結果 LPS與IEC-6共培養3h、5h、7h時,隨著LPS濃度的增加,IEC-6的增殖率逐漸降低.共培養3h和5h時,B組、C組、D組、E組和F組增殖率分彆與A組比較,差異均無統計學意義(P均>0.05);共培養7h時,B組、C組、D組增殖率與A組比較,差異均無統計學意義(P均>0.05),E組、F組增殖率與A組相比,差異有統計學意義(t=4.216,P=0.014;t=14.991,P=0.000).E組和F組共培養7h時的細胞增殖率較共培養5h時降低,差異均有統計學意義(t=2.576,P=0.033;t2.975,P=0.018);而共培養5h時E組與A組比較,差異無統計學意義(P>0.05).LPS與IEC-6共培養3h、5h和7h時,B組、C組、D組、E組和F組共培養上清中IL-6水平與A組比較均有升高,差異均有統計學意義(P均<0.01),其中各時間點E組IL-6水平均最高.E組LPS與IEC-6共培養5h時IL-6水平齣現峰值.LPS與IEC-6共培養3h、5h和7h時,B組、C組、D組、E組、F組共培養上清中IL-1β、TNF-α水平與A組比較,差異均無統計學意義(P均>0.05).結論 在一定濃度、一定培養時間範圍內,隨LPS濃度的增加,IEC-6增殖率呈下降趨勢.0~10.0 mg/L的LPS與IEC-6共培養,可刺激IEC-6分泌IL-6.5.0 mg/L的LPS與IEC-6共培養5h,其培養上清中IL-6水平最高.LPS對IEC-6培養上清中TNF-α和IL-1β水平無影響.5.0 mg/L的LPS與IEC-6共培養5h可製作較好的IEC-6炎性模型.
목적 탐토불동농도지다당(LPS)대대서장상피세포(IEC-6)증식화백세포개소(IL)-6、IL-1β급종류배사인자-α(TNF-α)분비적영향.방법 체외배양적IEC-6분위정상대조조(0 mg/L,A조)、0.1 mg/L조(B조)、0.5 mg/L조(C조)、1.0 mg/L조(D조)、5.0 mg/L조(E조)화10.0 mg/L조(F조),분별용대응농도적LPS여세포공배양3h、5h、7h후,새서람(MTT)비색법검측세포증식솔,매련면역흡부시험(ELISA)검측세포배양상청중IL-6、IL-1β급TNF-α적질량농도.결과 LPS여IEC-6공배양3h、5h、7h시,수착LPS농도적증가,IEC-6적증식솔축점강저.공배양3h화5h시,B조、C조、D조、E조화F조증식솔분별여A조비교,차이균무통계학의의(P균>0.05);공배양7h시,B조、C조、D조증식솔여A조비교,차이균무통계학의의(P균>0.05),E조、F조증식솔여A조상비,차이유통계학의의(t=4.216,P=0.014;t=14.991,P=0.000).E조화F조공배양7h시적세포증식솔교공배양5h시강저,차이균유통계학의의(t=2.576,P=0.033;t2.975,P=0.018);이공배양5h시E조여A조비교,차이무통계학의의(P>0.05).LPS여IEC-6공배양3h、5h화7h시,B조、C조、D조、E조화F조공배양상청중IL-6수평여A조비교균유승고,차이균유통계학의의(P균<0.01),기중각시간점E조IL-6수평균최고.E조LPS여IEC-6공배양5h시IL-6수평출현봉치.LPS여IEC-6공배양3h、5h화7h시,B조、C조、D조、E조、F조공배양상청중IL-1β、TNF-α수평여A조비교,차이균무통계학의의(P균>0.05).결론 재일정농도、일정배양시간범위내,수LPS농도적증가,IEC-6증식솔정하강추세.0~10.0 mg/L적LPS여IEC-6공배양,가자격IEC-6분비IL-6.5.0 mg/L적LPS여IEC-6공배양5h,기배양상청중IL-6수평최고.LPS대IEC-6배양상청중TNF-α화IL-1β수평무영향.5.0 mg/L적LPS여IEC-6공배양5h가제작교호적IEC-6염성모형.
Objective To investigate the effects of lipopolysaccharides (LPS) in different concentrations on the proliferation and interleukin(IL)-6,IL-1 β and tumor necrosis factor-α(TNF-o) secretion of intestinal epithelial cells (IEC-6) of rats in vitro.Methods IEC-6 of rats were divided into normal group (0 mg/L,group A),0.1 mg/L group (group B),0.5 mg/L group (group C),1.0 mg/L group (group D),5.0 mg/L group (group E) and 10.0 mg/L group(group F).Different groups cells were exposed to LPS with different concentrations for 3 h,5 h and 7 h.Thiazolyl blue(MTT) was performed to investigate the proliferation of IEC-6.The concentrations of IL-6,IL-1 β and TNF-α in culture supernatant were detected by enzyme linked immunosorbent assay(ELISA).Results The proliferation rate of IEC-6 was gradually lower while the concentration of LPS increased.After co-culture with LPS 3h and 5 h,the proliferation rates of group B,group C,group D,group E and group F had no significant difference with those of group A (all P > 0.05);after co-culture with LPS 7 h,the proliferation rates of group B,group C,and group D had no significant difference with those of group A (all P > 0.05);the proliferation rates of group E and group F had significant difference with those of group A(t =4.216,P =0.014;t =14.991,P =0.000).The proliferation rates of group E and group F were lower after co-culture with LPS 5 h than 7 h,and there were significant differences (t =2.576,P =0.033;t =2.975,P =0.018);but there was no significant differences between group E and group A after co-culture with LPS 5 h (P > 0.05).Group B,group C,group D,group E and group F all had a significant higher level of IL-6 than group A after co-culture with LPS 3 h,5 h and 7 h(all P <0.01).In addition,group E had the highest level of IL-6 at all time points.And the peak level of IL-6 rose after co-culture with LPS 5 h.The TNF-α level and IL-1 β level of group B,group C,group D,group E and group F all had no significant differences than that of group A after co-culture with LPS 3 h,5 h and 7 h (all P > 0.05).Conclusions In a certain concentration,incubation time range,the proliferation rates of IEC-6 cells were gradually lower while the concentration of LPS increased.Co-cultured IEC-6 cells with LPS(0-10.0 mg/L) can stimulate them secrete to IL-6.The highest level of IL-6 was of group E after 5 h co-culture.LPS had no effects on TNF-α and IL-1 β level of IEC-6 cells cultural supernatant.So 5.0 mg/L concentration of LPS stimulating IEC-6 cells for 5 h can be chosen to build the IEC-6 inflammatory models.