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
274-278
,共5页
姜洪娟%张玄%迟翔宇%王建平%王静%窦建明
薑洪娟%張玄%遲翔宇%王建平%王靜%竇建明
강홍연%장현%지상우%왕건평%왕정%두건명
哮喘%蛋白质组学%糖皮质激素
哮喘%蛋白質組學%糖皮質激素
효천%단백질조학%당피질격소
Asthma%Proteomics%Glucocorticoid
目的 比较分析支气管哮喘(简称哮喘)患者吸入糖皮质激素(ICS)治疗前后及健康人的血清蛋白质组,筛选在哮喘发病机制及ICS治疗中起关键作用的靶点蛋白质.方法 选择2011年6月至2012年9月就诊于山东省立医院的慢性持续期哮喘患者30例(哮喘组)及健康志愿者30例(对照组),哮喘组给予规范的ICS治疗8周,收集哮喘组治疗前后及对照组的血清,应用比较蛋白质组学技术,筛选并鉴定差异蛋白质.对ICS治疗前后差异显著的蛋白质进行Western blot验证及ELISA检测,分析其在各组血清中的浓度变化,并与哮喘患者的IgE、嗜酸粒细胞(EOS)、中性粒细胞百分比(NEUT％)、FEV1占预计值％(FEV1％)行相关分析.结果 哮喘组共鉴定出11个差异表达的蛋白质,其中热休克蛋白70(HSP70)、嗜酸粒细胞趋化蛋白(Eotaxin)、维生素D结合蛋白(VDBP)在ICS治疗前后蛋白丰度差异有统计学意义(均P＜0.05).Western blot验证了HSP70、Eotaxin和VDBP在各组血清中的表达变化与蛋白质组学结果相一致.ELISA数据显示,哮喘组治疗前血清HSP70和Eotaxin水平[分别为(439±103) ng/L和(183±79) ng/L],明显高于对照组[(209±58) ng/L和(91±46)ng/L](t值分别为5.281、4.972,均P＜0.01),哮喘组治疗后血清HSP70和Eotaxin水平[分别为(247±96) ng/L和(105±58)ng/L]明显低于治疗前(t值分别为4.157、3.892,均P＜0.01)];哮喘组治疗前血清VDBP水平[(318±115)mg/L]明显低于对照组[(541±98)mg/L](f=3.878,P＜0.01),哮喘组治疗后血清VDBP水平[(479±132) mg/L]明显高于治疗前(t=3.572,P＜0.01).相关分析显示,HSP70与哮喘患者IgE、NEUT％呈正相关,与FEV1％呈负相关(r值分别为0.568、0.613、-0.516,均P＜0.01);Eotaxin与哮喘患者IgE、EOS呈正相关,与FEV1％呈负相关(r值分别为0.752、0.826、-0.618,均P＜0.01);VDBP与哮喘患者NEUT％呈负相关,与FEV1％呈正相关(r值分别为-0.537、0.426,均P＜0.05).结论 HSP70、Eotaxin、VDBP不仅参与哮喘的发病机制,而且有可能成为ICS治疗的新靶点.
目的 比較分析支氣管哮喘(簡稱哮喘)患者吸入糖皮質激素(ICS)治療前後及健康人的血清蛋白質組,篩選在哮喘髮病機製及ICS治療中起關鍵作用的靶點蛋白質.方法 選擇2011年6月至2012年9月就診于山東省立醫院的慢性持續期哮喘患者30例(哮喘組)及健康誌願者30例(對照組),哮喘組給予規範的ICS治療8週,收集哮喘組治療前後及對照組的血清,應用比較蛋白質組學技術,篩選併鑒定差異蛋白質.對ICS治療前後差異顯著的蛋白質進行Western blot驗證及ELISA檢測,分析其在各組血清中的濃度變化,併與哮喘患者的IgE、嗜痠粒細胞(EOS)、中性粒細胞百分比(NEUT％)、FEV1佔預計值％(FEV1％)行相關分析.結果 哮喘組共鑒定齣11箇差異錶達的蛋白質,其中熱休剋蛋白70(HSP70)、嗜痠粒細胞趨化蛋白(Eotaxin)、維生素D結閤蛋白(VDBP)在ICS治療前後蛋白豐度差異有統計學意義(均P＜0.05).Western blot驗證瞭HSP70、Eotaxin和VDBP在各組血清中的錶達變化與蛋白質組學結果相一緻.ELISA數據顯示,哮喘組治療前血清HSP70和Eotaxin水平[分彆為(439±103) ng/L和(183±79) ng/L],明顯高于對照組[(209±58) ng/L和(91±46)ng/L](t值分彆為5.281、4.972,均P＜0.01),哮喘組治療後血清HSP70和Eotaxin水平[分彆為(247±96) ng/L和(105±58)ng/L]明顯低于治療前(t值分彆為4.157、3.892,均P＜0.01)];哮喘組治療前血清VDBP水平[(318±115)mg/L]明顯低于對照組[(541±98)mg/L](f=3.878,P＜0.01),哮喘組治療後血清VDBP水平[(479±132) mg/L]明顯高于治療前(t=3.572,P＜0.01).相關分析顯示,HSP70與哮喘患者IgE、NEUT％呈正相關,與FEV1％呈負相關(r值分彆為0.568、0.613、-0.516,均P＜0.01);Eotaxin與哮喘患者IgE、EOS呈正相關,與FEV1％呈負相關(r值分彆為0.752、0.826、-0.618,均P＜0.01);VDBP與哮喘患者NEUT％呈負相關,與FEV1％呈正相關(r值分彆為-0.537、0.426,均P＜0.05).結論 HSP70、Eotaxin、VDBP不僅參與哮喘的髮病機製,而且有可能成為ICS治療的新靶點.
목적 비교분석지기관효천(간칭효천)환자흡입당피질격소(ICS)치료전후급건강인적혈청단백질조,사선재효천발병궤제급ICS치료중기관건작용적파점단백질.방법 선택2011년6월지2012년9월취진우산동성립의원적만성지속기효천환자30례(효천조)급건강지원자30례(대조조),효천조급여규범적ICS치료8주,수집효천조치료전후급대조조적혈청,응용비교단백질조학기술,사선병감정차이단백질.대ICS치료전후차이현저적단백질진행Western blot험증급ELISA검측,분석기재각조혈청중적농도변화,병여효천환자적IgE、기산립세포(EOS)、중성립세포백분비(NEUT％)、FEV1점예계치％(FEV1％)행상관분석.결과 효천조공감정출11개차이표체적단백질,기중열휴극단백70(HSP70)、기산립세포추화단백(Eotaxin)、유생소D결합단백(VDBP)재ICS치료전후단백봉도차이유통계학의의(균P＜0.05).Western blot험증료HSP70、Eotaxin화VDBP재각조혈청중적표체변화여단백질조학결과상일치.ELISA수거현시,효천조치료전혈청HSP70화Eotaxin수평[분별위(439±103) ng/L화(183±79) ng/L],명현고우대조조[(209±58) ng/L화(91±46)ng/L](t치분별위5.281、4.972,균P＜0.01),효천조치료후혈청HSP70화Eotaxin수평[분별위(247±96) ng/L화(105±58)ng/L]명현저우치료전(t치분별위4.157、3.892,균P＜0.01)];효천조치료전혈청VDBP수평[(318±115)mg/L]명현저우대조조[(541±98)mg/L](f=3.878,P＜0.01),효천조치료후혈청VDBP수평[(479±132) mg/L]명현고우치료전(t=3.572,P＜0.01).상관분석현시,HSP70여효천환자IgE、NEUT％정정상관,여FEV1％정부상관(r치분별위0.568、0.613、-0.516,균P＜0.01);Eotaxin여효천환자IgE、EOS정정상관,여FEV1％정부상관(r치분별위0.752、0.826、-0.618,균P＜0.01);VDBP여효천환자NEUT％정부상관,여FEV1％정정상관(r치분별위-0.537、0.426,균P＜0.05).결론 HSP70、Eotaxin、VDBP불부삼여효천적발병궤제,이차유가능성위ICS치료적신파점.
Objective To screen the biomarkers which may play important roles in the pathogenesis and therapy of asthma by using serum comparative proteomics.Methods From June 2011 to September 2012,30 chronic persistent asthmatic patients (asthma group) and 30 healthy controls (control group) were selected for study in our hospital.All the asthmatic patients were given 8 week-treatment with inhaled glucocorticoids(ICS).Then comparative proteomics were employed to identify differential proteins in serum samples from the control group,the asthma pre-treatment group and the asthma post-treatment group.The differential proteins which had significant differences before and after ICS therapy were selected for Western blot analysis and ELISA detection.Besides,the correlation between the differential proteins and IgE,eosinophils (EOS),neutrophil percentage (NEUT％),and FEV1 ％ were analyzed.Results Eleven differential proteins were identified.Among them,heat shock protein 70 (HSP70),eosinophil chemotactic protein(Eotaxin) and vitamin D binding protein(VDBP) had significant differences in protein abundance before and after treatment.The differential expression of the 3 proteins in each group was confirmed by Western blot.ELISA data showed that the serum levels of HSP70 and Eotaxin were significantly higher in the asthma pre-treatment group[(439 ± 103)ng/L,(183 ±79)ng/L] than those in the control group [(209 ± 58)ng/L,(91 ± 46) ng/L] (t =5.281,4.972,all P ＜ 0.01),but significantly lower in the asthma post-treatment group[(247 ± 96)ng/L,(105 ± 58)ng/L] than those in the pre-treatment group(t =4.157,3.892,all P ＜ 0.01).However,the serum level of VDBP was significantly lower in the asthmatics pre-treatment group [(318 ± 115)mg/L] than that in the control group[(541 ± 98)mg/L] (t =3.878,P ＜ 0.01),but significantly higher in the asthma post-treatment group [(479 ± 132)mg/L] than that in the pre-treatment group(t =3.572,P ＜ 0.01).The correlation analysis showed that HSP70 was correlated positively with IgE and NEUT％,but negatively with FEV1 ％ (r =0.568,0.613,-0.516,all P ＜ 0.01).Eotaxin was correlated positively with IgE and EOS,but negatively with FEV1％ (r =0.752,0.826,-0.618,all P ＜0.01).VDBP was correlated negatively with NEUT％,but positively with FEV1 ％ (r =-0.537,0.426,all P ＜ 0.05).Conclusions HSP70,Eotaxin,and VDBP may participate in the pathogenesis of asthma,and may become the potential targets for ICS therapy.
