中华结核和呼吸杂志
中華結覈和呼吸雜誌
중화결핵화호흡잡지
Chinese Journal of Tuberculosis and Respiratory Diseases
2014年
4期
250-254
,共5页
曾晓丽%刘晓菊%包海荣%张艺%王小虎%施凯%庞琪
曾曉麗%劉曉菊%包海榮%張藝%王小虎%施凱%龐琪
증효려%류효국%포해영%장예%왕소호%시개%방기
肺疾病,慢性阻塞性%Toll样受体4%髓样分化因子88%巨噬细胞
肺疾病,慢性阻塞性%Toll樣受體4%髓樣分化因子88%巨噬細胞
폐질병,만성조새성%Toll양수체4%수양분화인자88%거서세포
Pulmonary disease,chronic obstructive%Toll-like receptor 4%Myeloid differentiation factor 88%Macrophages
目的 探讨萝卜硫素对慢性阻塞性肺疾病(简称慢阻肺)患者Toll样受体4(TLR4)、髓样分化因子88(MyD88)和下游炎性因子的影响.方法 选择2012年1月至2013年3月兰州大学第一医院慢阻肺稳定期患者32例为慢阻肺组,同期健康体检者30名为非慢阻肺组,分离外周血单核细胞,诱导形成单核细胞源性巨噬细胞(MDM).将慢阻肺组MDM分为空白对照组、脂多糖组、萝卜硫素组和萝卜硫素联合脂多糖组(简称联合组),非慢阻肺组不用任何药物干预,每组3×106个细胞.实时荧光定量PCR和免疫印迹法检测MDM中TLR4、MyD88 mRNA和蛋白表达,酶联免疫吸附试验检测细胞培养上清液中肿瘤坏死因子-α(TNF-α)和IL-6含量.多组间比较采用单因素方差分析和LSD-t检验.结果 空白对照组TLR4、MyD88 mRNA和蛋白表达(3.7±0.5、1.9±0.4和0.45±0.18、1.11 ±0.65)及细胞培养上清液中TNF-α和IL-6浓度[(31±4)和(43 ±5) μg/L]显著高于非慢阻肺组[(1.00、1.00和0.26 ±0.14、0.58 ±0.40、(19±2)和(29±4)μg/L],差异均有统计学意义(t值为2.19～12.11,P＜0.05或P＜0.01);脂多糖组上述指标[5.5±1.1、3.4±1.6和0.65±0.20、1.66±0.64、(47±4)和(54 ±5)μg/L]显著高于空白对照组,差异均有统计学意义(t值为2.39～11.9,P＜0.05或P＜0.01);萝卜硫素组上述指标[2.2±0.4、1.0±0.6和0.25±0.09、0.62±0.34、(20±3)和(27±4) μg/L]显著低于空白对照组,差异均有统计学意义(t值为2.13 ～8.46,P＜0.05或P＜0.01);联合组上述指标[3.2±0.5、1.5±0.8和0.33 ±0.11、0.77 ±0.25、(31±3)和(33±4) μg/L]显著低于脂多糖组,差异均有统计学意义(t值为3.87 ～ 12.24,均P＜0.01).结论 慢阻肺患者的巨噬细胞TLR4/MyD88信号通路活化,下游炎性因子分泌增加.萝卜硫素可抑制TLR4/MyD88通路,减少下游炎性因子的释放,可能对慢阻肺患者有抗炎作用.
目的 探討蘿蔔硫素對慢性阻塞性肺疾病(簡稱慢阻肺)患者Toll樣受體4(TLR4)、髓樣分化因子88(MyD88)和下遊炎性因子的影響.方法 選擇2012年1月至2013年3月蘭州大學第一醫院慢阻肺穩定期患者32例為慢阻肺組,同期健康體檢者30名為非慢阻肺組,分離外週血單覈細胞,誘導形成單覈細胞源性巨噬細胞(MDM).將慢阻肺組MDM分為空白對照組、脂多糖組、蘿蔔硫素組和蘿蔔硫素聯閤脂多糖組(簡稱聯閤組),非慢阻肺組不用任何藥物榦預,每組3×106箇細胞.實時熒光定量PCR和免疫印跡法檢測MDM中TLR4、MyD88 mRNA和蛋白錶達,酶聯免疫吸附試驗檢測細胞培養上清液中腫瘤壞死因子-α(TNF-α)和IL-6含量.多組間比較採用單因素方差分析和LSD-t檢驗.結果 空白對照組TLR4、MyD88 mRNA和蛋白錶達(3.7±0.5、1.9±0.4和0.45±0.18、1.11 ±0.65)及細胞培養上清液中TNF-α和IL-6濃度[(31±4)和(43 ±5) μg/L]顯著高于非慢阻肺組[(1.00、1.00和0.26 ±0.14、0.58 ±0.40、(19±2)和(29±4)μg/L],差異均有統計學意義(t值為2.19～12.11,P＜0.05或P＜0.01);脂多糖組上述指標[5.5±1.1、3.4±1.6和0.65±0.20、1.66±0.64、(47±4)和(54 ±5)μg/L]顯著高于空白對照組,差異均有統計學意義(t值為2.39～11.9,P＜0.05或P＜0.01);蘿蔔硫素組上述指標[2.2±0.4、1.0±0.6和0.25±0.09、0.62±0.34、(20±3)和(27±4) μg/L]顯著低于空白對照組,差異均有統計學意義(t值為2.13 ～8.46,P＜0.05或P＜0.01);聯閤組上述指標[3.2±0.5、1.5±0.8和0.33 ±0.11、0.77 ±0.25、(31±3)和(33±4) μg/L]顯著低于脂多糖組,差異均有統計學意義(t值為3.87 ～ 12.24,均P＜0.01).結論 慢阻肺患者的巨噬細胞TLR4/MyD88信號通路活化,下遊炎性因子分泌增加.蘿蔔硫素可抑製TLR4/MyD88通路,減少下遊炎性因子的釋放,可能對慢阻肺患者有抗炎作用.
목적 탐토라복류소대만성조새성폐질병(간칭만조폐)환자Toll양수체4(TLR4)、수양분화인자88(MyD88)화하유염성인자적영향.방법 선택2012년1월지2013년3월란주대학제일의원만조폐은정기환자32례위만조폐조,동기건강체검자30명위비만조폐조,분리외주혈단핵세포,유도형성단핵세포원성거서세포(MDM).장만조폐조MDM분위공백대조조、지다당조、라복류소조화라복류소연합지다당조(간칭연합조),비만조폐조불용임하약물간예,매조3×106개세포.실시형광정량PCR화면역인적법검측MDM중TLR4、MyD88 mRNA화단백표체,매련면역흡부시험검측세포배양상청액중종류배사인자-α(TNF-α)화IL-6함량.다조간비교채용단인소방차분석화LSD-t검험.결과 공백대조조TLR4、MyD88 mRNA화단백표체(3.7±0.5、1.9±0.4화0.45±0.18、1.11 ±0.65)급세포배양상청액중TNF-α화IL-6농도[(31±4)화(43 ±5) μg/L]현저고우비만조폐조[(1.00、1.00화0.26 ±0.14、0.58 ±0.40、(19±2)화(29±4)μg/L],차이균유통계학의의(t치위2.19～12.11,P＜0.05혹P＜0.01);지다당조상술지표[5.5±1.1、3.4±1.6화0.65±0.20、1.66±0.64、(47±4)화(54 ±5)μg/L]현저고우공백대조조,차이균유통계학의의(t치위2.39～11.9,P＜0.05혹P＜0.01);라복류소조상술지표[2.2±0.4、1.0±0.6화0.25±0.09、0.62±0.34、(20±3)화(27±4) μg/L]현저저우공백대조조,차이균유통계학의의(t치위2.13 ～8.46,P＜0.05혹P＜0.01);연합조상술지표[3.2±0.5、1.5±0.8화0.33 ±0.11、0.77 ±0.25、(31±3)화(33±4) μg/L]현저저우지다당조,차이균유통계학의의(t치위3.87 ～ 12.24,균P＜0.01).결론 만조폐환자적거서세포TLR4/MyD88신호통로활화,하유염성인자분비증가.라복류소가억제TLR4/MyD88통로,감소하유염성인자적석방,가능대만조폐환자유항염작용.
Objective To explore the effects of sulforaphane on Toll-like receptor 4 (TLR4)/ myeloid differentiation factor 88 (MyD88) pathway and its downstream inflammatory cytokines in patients with chronic obstructive pulmonary disease (COPD).Methods From Jan.2012 to Mar.2013,thirty-two stable COPD patients and thirty healthy donors (non-COPD group) from the First Hospital of Lanzhou University were recruited.The peripheral blood monocytes were isolated and induced to macrophages (monocyte-derived macrophages,MDMs).The MDMs of COPD patients were divided into a blank control group,a LPS group,a sulforaphane group,a sulforaphane and LPS group (combined group),while the MDMs from the non-COPD group received no drug intervention.The number of cells in each group was 3 ×106.The mRNA and protein expression of TLR4 and MyD88 were measured with real-time PCR and Western blot.The TNF-α and IL-6 levels in the culture supernatant were measured with ELISA.Oneway ANOVA and LSD-t test were used for statistical analysis.Results The levels of mRNA and protein of TLR4 and MyD88 and the contents of TNF-α and IL-6 in the culture supernatant were higher in the blank control group [3.7 ±0.5,1.9±0.4,0.45 ±0.18,1.11 ±0.65,(31 ±4) and (43 ±5) μg/L] than those in the nonCOPDgroup [1.00,1.00,0.26±0.14,0.58±0.40,(19±2) and (29±4) μg/L] (t=2.19-12.11,P ＜0.05 or P ＜0.01).After LPS treatment (LPS group),the above parameters [5.5 ± 1.1,3.4 ± 1.6,0.65 ± 0.20,1.66 ± 0.64,(47 ± 4) and (54 ± 5) μg/L] were increased as compared to those in the blank control group (t =2.39-11.9,P ＜ 0.05 or P ＜ 0.01),but after sulforaphane treatment (Sulforaphane group),these parameters [2.2 ± 0.4,1.0 ± 0.6,0.25 ± 0.09,0.62 ± 0.34,(20 ± 3) and (27 ±4) μg/L] were decreased as compared to those in the blank control group (t =2.13-8.46,P ＜ 0.05 or P ＜ 0.01).Similarly,these parameters in the combined group [3.2 ± 0.5,1.5 ± 0.8,0.33 ± 0.11,0.77 ±0.25,(31 ±3) and (33 ±4) μg/L] were also remarkably decreased as compared to those in the LPS group (t =3.87-12.24,all P＜0.01).Conclusions The TLR4/MyD88 pathway was activated and its downstream inflammatory cytokines were increased in macrophages from COPD patients.Sulforaphane could inhibit the TLR4/MyD88 pathway and reduce the releasing of downstream inflammatory cytokines,suggesting that sulforaphane may have an anti-inflammatory effect in COPD.
