中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2014年
3期
218-222
,共5页
肖志诚%张晶%张俊蒙%王绿娅%杜杰
肖誌誠%張晶%張俊矇%王綠婭%杜傑
초지성%장정%장준몽%왕록아%두걸
心肌梗死%巨噬细胞%趋化因子%吞噬作用
心肌梗死%巨噬細胞%趨化因子%吞噬作用
심기경사%거서세포%추화인자%탄서작용
Myocardial infarction%Macrophage%Chemochine%Phagocytosis
目的 探讨趋化因子配体16(CXCL16)在心肌梗死早期是否升高及其在体外对巨噬细胞吞噬功能的影响.方法 野生雄性C57 BL/6J小鼠40只按随机数字表随机分为心肌梗死组和假手术组,每组各20只;心肌梗死组开胸结扎小鼠冠脉前降支,假手术组只给予开胸处理;术后立即行心电图检查;于3d时每组各处死10只小鼠,ELISA检测外周血清中CXCL16的表达;心脏组织病理切片,HE染色观察心脏组织的结构改变和炎症浸润情况;免疫组化检测CXCL16的表达;其余每组各10只小鼠于28 d时显微超声检测小鼠心脏结构及功能;心脏组织病理切片,Masson三原色法检测心脏胶原沉积;HE染色观察心脏组织的结构改变和炎症浸润情况;免疫组化检测CXCL16的表达;体外实验分离小鼠骨髓单核细胞,刺激分化为巨噬细胞后转染CXCL16腺病毒及绿色荧光蛋白(GFP)对照,并与小鼠心脏细胞碎片共培养,流式细胞仪检测巨噬细胞吞噬功能.结果 与假手术组相比,心肌梗死手术后小鼠即刻出现心律加快及ST段抬高;3d时外周血CXCL16水平上调[(1 079±176)pg/ml比(611 ±37) pg/ml,P=0.032],梗死区心肌细胞坏死,大量炎症细胞浸润;免疫组化染色可见CXCL16表达上调;28 d时心肌梗死组左室射血分数降低(13.29％±2.83％比48.92％±5.46％,P＜0.叭),梗死区胶原沉积,炎症消退,免疫组化见CXCL16表达下降;流式细胞术检测可见巨噬细胞转染CXCL16腺病毒后吞噬功能增强(17.11％ ±0.87％比7.91％±0.71％,P＜0.01).结论 小鼠心肌梗死早期CXCL16表达上调,炎症细胞浸润,体外CXCL16在巨噬细胞过表达能促进其对细胞碎片的吞噬作用.
目的 探討趨化因子配體16(CXCL16)在心肌梗死早期是否升高及其在體外對巨噬細胞吞噬功能的影響.方法 野生雄性C57 BL/6J小鼠40隻按隨機數字錶隨機分為心肌梗死組和假手術組,每組各20隻;心肌梗死組開胸結扎小鼠冠脈前降支,假手術組隻給予開胸處理;術後立即行心電圖檢查;于3d時每組各處死10隻小鼠,ELISA檢測外週血清中CXCL16的錶達;心髒組織病理切片,HE染色觀察心髒組織的結構改變和炎癥浸潤情況;免疫組化檢測CXCL16的錶達;其餘每組各10隻小鼠于28 d時顯微超聲檢測小鼠心髒結構及功能;心髒組織病理切片,Masson三原色法檢測心髒膠原沉積;HE染色觀察心髒組織的結構改變和炎癥浸潤情況;免疫組化檢測CXCL16的錶達;體外實驗分離小鼠骨髓單覈細胞,刺激分化為巨噬細胞後轉染CXCL16腺病毒及綠色熒光蛋白(GFP)對照,併與小鼠心髒細胞碎片共培養,流式細胞儀檢測巨噬細胞吞噬功能.結果 與假手術組相比,心肌梗死手術後小鼠即刻齣現心律加快及ST段抬高;3d時外週血CXCL16水平上調[(1 079±176)pg/ml比(611 ±37) pg/ml,P=0.032],梗死區心肌細胞壞死,大量炎癥細胞浸潤;免疫組化染色可見CXCL16錶達上調;28 d時心肌梗死組左室射血分數降低(13.29％±2.83％比48.92％±5.46％,P＜0.叭),梗死區膠原沉積,炎癥消退,免疫組化見CXCL16錶達下降;流式細胞術檢測可見巨噬細胞轉染CXCL16腺病毒後吞噬功能增彊(17.11％ ±0.87％比7.91％±0.71％,P＜0.01).結論 小鼠心肌梗死早期CXCL16錶達上調,炎癥細胞浸潤,體外CXCL16在巨噬細胞過錶達能促進其對細胞碎片的吞噬作用.
목적 탐토추화인자배체16(CXCL16)재심기경사조기시부승고급기재체외대거서세포탄서공능적영향.방법 야생웅성C57 BL/6J소서40지안수궤수자표수궤분위심기경사조화가수술조,매조각20지;심기경사조개흉결찰소서관맥전강지,가수술조지급여개흉처리;술후립즉행심전도검사;우3d시매조각처사10지소서,ELISA검측외주혈청중CXCL16적표체;심장조직병리절편,HE염색관찰심장조직적결구개변화염증침윤정황;면역조화검측CXCL16적표체;기여매조각10지소서우28 d시현미초성검측소서심장결구급공능;심장조직병리절편,Masson삼원색법검측심장효원침적;HE염색관찰심장조직적결구개변화염증침윤정황;면역조화검측CXCL16적표체;체외실험분리소서골수단핵세포,자격분화위거서세포후전염CXCL16선병독급록색형광단백(GFP)대조,병여소서심장세포쇄편공배양,류식세포의검측거서세포탄서공능.결과 여가수술조상비,심기경사수술후소서즉각출현심률가쾌급ST단태고;3d시외주혈CXCL16수평상조[(1 079±176)pg/ml비(611 ±37) pg/ml,P=0.032],경사구심기세포배사,대량염증세포침윤;면역조화염색가견CXCL16표체상조;28 d시심기경사조좌실사혈분수강저(13.29％±2.83％비48.92％±5.46％,P＜0.팔),경사구효원침적,염증소퇴,면역조화견CXCL16표체하강;류식세포술검측가견거서세포전염CXCL16선병독후탄서공능증강(17.11％ ±0.87％비7.91％±0.71％,P＜0.01).결론 소서심기경사조기CXCL16표체상조,염증세포침윤,체외CXCL16재거서세포과표체능촉진기대세포쇄편적탄서작용.
Objective To explore whether chemokine CXCL16 is up-regulated after myocardial infarction and promotes the phagocytic activity of macrophage in vitro.Methods Forty wild-type mice were randomly separated into 2 groups (n =20 each).Group A had the ligation of left anterior descending coronary artery while group B underwent a sham operation.Electrocardiogram was used to assess whether the operation was successful or not.Three days after surgery,10 animals of each group were sacrificed and the serum level of CXCL16 was detected by enzyme-linked immunosorbent assay (ELISA).Twenty-eight days after surgery,cardiac function of the remaining mice was measured by small animal ultrasound.Then the animals were sacrificed.Hematoxylin and eosin (HE) staining and immunohistochemical staining of cardiac paraffin section were used to observe the inflammation and detect the expression of CXCL16 in cardiac tissue after myocardial infarction.To explore the function of CXCL16 in vitro,primary murine monocytes were separated from bone marrow,cultured to differentiate into macrophages and transfected with adenovirus vectors over-expressing CXCL16 or control adenovirus vectors.After stimulation by debris of cardiac cells,the phagocytic uptake by macrophages was evaluated by flow cytometry.Results The model of myocardial infarction was successfully established.ELISA showed that the serum level of CXCL16 was elevated 3 days after myocardial infarction [(1 079 ± 176) vs (611 ± 37) pg/ml,P =0.032].HE and immunohistochemical staining demonstrated that the infiltration of macrophages increased during an early stage of myocardial infarction and decreased at the late stage.The CXCL16 level was up-regulated 3 days after myocardial infarction and returned to normal level at Day 28.Furthermore,macrophages transfected with adenovirns over-expressing CXCL16 showed stronger phagocytic activity compared with control (17.11％±0.87％ vs 7.91％ ±0.71％,P＜0.01).Conclusion CXCL16 is up-regulated after myocardial infarction in mice.And an in vitro over-expression of CXCL16 promotes the macrophage phagocytosis of cardiac debris.
