中华老年医学杂志
中華老年醫學雜誌
중화노년의학잡지
Chinese Journal of Geriatrics
2013年
9期
996-1000
,共5页
刘志江%石蓓%许官学%赵然尊%沈长银%陈攀科
劉誌江%石蓓%許官學%趙然尊%瀋長銀%陳攀科
류지강%석배%허관학%조연존%침장은%진반과
受体,CXCR4%间质干细胞%颈动脉损伤
受體,CXCR4%間質榦細胞%頸動脈損傷
수체,CXCR4%간질간세포%경동맥손상
Receptors,CXCR4%Mesenchymal stem cells%Carotid artery injuries
目的 探讨基质细胞衍生因子受体(CXCR4)基因修饰的骨髓间充质干细胞(BMSCs)移植对大鼠颈动脉损伤后修复的影响. 方法 体外培养和扩增BMSCs,用携带CXCR4的慢病毒质粒(Lenti vector)转染BMSCs,获得CXCR4修饰的BMSCs(CXCR4-BMSCs),免疫印迹(Westernblot)法检测CXCR4的表达情况.建立大鼠颈动脉球囊损伤模型,随机分成CXCR4转染BMSCs移植组(CXCR4-BMSCs,12只),空病毒转染BMSCs组(BMSCs组,12只)及磷酸盐缓冲液(PBS)移植对照组(对照组,12只).细胞移植后2周取血管组织分别行绿色荧光蛋白(GFP)荧光检测BMSCs归巢情况,免疫荧光染色检测血管内膜血小板-内皮细胞黏附分子(CD31)的表达,细胞移植后4周免疫组织化学染色检测增殖细胞核抗原(PCNA)的表达及苏木精-伊红(HE)染色检测血管形态学变化. 结果 BMSCs转染Lenti-CXCR4后CXCR4蛋白水平与转染Lenti-vector和未转染的BMSCs比较呈高表达(P＜0.05).CXCR4-BMSCs组和BMSCs组血管内膜均有GFP阳性细胞归巢,CXCR4-BMSCs组(58.8±4.4)％较BMSCs组(36.2±5.0)％增加(P＜0.05).CXCR4-BMSCs组CD31表达(58.8±4.3)％高于BMSCs组(28.8±4.2)％(P＜0.05).PCNA的表达CXCR4-BMSCs组(21.0±4.2)％与BMSCs组(36.5±4.9)％较对照组(78.3±3.5)％均降低(P＜0.05),CXCR4-BMSCs较BMSCs组降低(P＜0.05).CXCR4-BMSCs组与BMSCs组新生内膜面积[分别为(0.205±0.018)mm2、(0.323±0.071) mm2]、新生内膜/中膜面积[分别为(1.039±0.123)、(1.660±0.404)]均较对照组减轻[(0.536±0.054) mm2、(2.460±0.328),P＜0.05]；CXCR4-BMSCs较BMSCs组也降低(P＜0.05). 结论 基因修饰可提高BMSCs的CXCR4表达,CXCR4-BMSCs较单纯BMSCs移植更能增加BMSCs归巢及促进颈动脉球囊损伤后早期再内皮化并减轻血管再狭窄.
目的 探討基質細胞衍生因子受體(CXCR4)基因脩飾的骨髓間充質榦細胞(BMSCs)移植對大鼠頸動脈損傷後脩複的影響. 方法 體外培養和擴增BMSCs,用攜帶CXCR4的慢病毒質粒(Lenti vector)轉染BMSCs,穫得CXCR4脩飾的BMSCs(CXCR4-BMSCs),免疫印跡(Westernblot)法檢測CXCR4的錶達情況.建立大鼠頸動脈毬囊損傷模型,隨機分成CXCR4轉染BMSCs移植組(CXCR4-BMSCs,12隻),空病毒轉染BMSCs組(BMSCs組,12隻)及燐痠鹽緩遲液(PBS)移植對照組(對照組,12隻).細胞移植後2週取血管組織分彆行綠色熒光蛋白(GFP)熒光檢測BMSCs歸巢情況,免疫熒光染色檢測血管內膜血小闆-內皮細胞黏附分子(CD31)的錶達,細胞移植後4週免疫組織化學染色檢測增殖細胞覈抗原(PCNA)的錶達及囌木精-伊紅(HE)染色檢測血管形態學變化. 結果 BMSCs轉染Lenti-CXCR4後CXCR4蛋白水平與轉染Lenti-vector和未轉染的BMSCs比較呈高錶達(P＜0.05).CXCR4-BMSCs組和BMSCs組血管內膜均有GFP暘性細胞歸巢,CXCR4-BMSCs組(58.8±4.4)％較BMSCs組(36.2±5.0)％增加(P＜0.05).CXCR4-BMSCs組CD31錶達(58.8±4.3)％高于BMSCs組(28.8±4.2)％(P＜0.05).PCNA的錶達CXCR4-BMSCs組(21.0±4.2)％與BMSCs組(36.5±4.9)％較對照組(78.3±3.5)％均降低(P＜0.05),CXCR4-BMSCs較BMSCs組降低(P＜0.05).CXCR4-BMSCs組與BMSCs組新生內膜麵積[分彆為(0.205±0.018)mm2、(0.323±0.071) mm2]、新生內膜/中膜麵積[分彆為(1.039±0.123)、(1.660±0.404)]均較對照組減輕[(0.536±0.054) mm2、(2.460±0.328),P＜0.05]；CXCR4-BMSCs較BMSCs組也降低(P＜0.05). 結論 基因脩飾可提高BMSCs的CXCR4錶達,CXCR4-BMSCs較單純BMSCs移植更能增加BMSCs歸巢及促進頸動脈毬囊損傷後早期再內皮化併減輕血管再狹窄.
목적 탐토기질세포연생인자수체(CXCR4)기인수식적골수간충질간세포(BMSCs)이식대대서경동맥손상후수복적영향. 방법 체외배양화확증BMSCs,용휴대CXCR4적만병독질립(Lenti vector)전염BMSCs,획득CXCR4수식적BMSCs(CXCR4-BMSCs),면역인적(Westernblot)법검측CXCR4적표체정황.건립대서경동맥구낭손상모형,수궤분성CXCR4전염BMSCs이식조(CXCR4-BMSCs,12지),공병독전염BMSCs조(BMSCs조,12지)급린산염완충액(PBS)이식대조조(대조조,12지).세포이식후2주취혈관조직분별행록색형광단백(GFP)형광검측BMSCs귀소정황,면역형광염색검측혈관내막혈소판-내피세포점부분자(CD31)적표체,세포이식후4주면역조직화학염색검측증식세포핵항원(PCNA)적표체급소목정-이홍(HE)염색검측혈관형태학변화. 결과 BMSCs전염Lenti-CXCR4후CXCR4단백수평여전염Lenti-vector화미전염적BMSCs비교정고표체(P＜0.05).CXCR4-BMSCs조화BMSCs조혈관내막균유GFP양성세포귀소,CXCR4-BMSCs조(58.8±4.4)％교BMSCs조(36.2±5.0)％증가(P＜0.05).CXCR4-BMSCs조CD31표체(58.8±4.3)％고우BMSCs조(28.8±4.2)％(P＜0.05).PCNA적표체CXCR4-BMSCs조(21.0±4.2)％여BMSCs조(36.5±4.9)％교대조조(78.3±3.5)％균강저(P＜0.05),CXCR4-BMSCs교BMSCs조강저(P＜0.05).CXCR4-BMSCs조여BMSCs조신생내막면적[분별위(0.205±0.018)mm2、(0.323±0.071) mm2]、신생내막/중막면적[분별위(1.039±0.123)、(1.660±0.404)]균교대조조감경[(0.536±0.054) mm2、(2.460±0.328),P＜0.05]；CXCR4-BMSCs교BMSCs조야강저(P＜0.05). 결론 기인수식가제고BMSCs적CXCR4표체,CXCR4-BMSCs교단순BMSCs이식경능증가BMSCs귀소급촉진경동맥구낭손상후조기재내피화병감경혈관재협착.
Objective To investigate the effect of transplantation of CXC receptor 4 (CXCR4)gene-modified bone marrow mesenchymal stem cells (BMSCs) on repairment of carotid injure in rats.Methods BMSCs were cultured and transfected with lentivirus vector carrying CXCR4 gene to generate CXCR4 gene-modified BMSCs (CXCR4-BMSCs).CXCR4 expression was detected by Western blot.Rat model of carotid artery balloon injury was established.Rats were randomly divided into the PBS control group (n=12),CXCR4-BMSCs group (n=12) and BMSCs group (n=12).Two weeks after transplantation,the injured arteries were obtained.The homing of BMSCs was detected by immunofluorescence with green fluorescent protein (GFP).Platelet endothelial cell adhesion molecule (CD31) expression was detected by immunofluorescence staining.At 4 weeks after transplantation,proliferating cell nuclear antigen (PCNA) expression was determined by immunohistochemical staining,and the vascular morphological changes were observed by hematoxylineosin staining (HE).Results Compared with the control and BMSCs groups,the protein level of CXCR4 was increased in CXCR4-BMSCs group (both P＜0.05).The percentage of GFP-positive cells homing were much more in CXCR4-BMSCs group than in BMSCs group [(58.8±4.4)％ vs.(36.2±5.0) ％,P＜0.05].The CD31 expression were higher in CXCR4-BMSCs group than in BMSCs group [(58.8±4.3)％ vs.(28.8±4.2)％,P＜0.05].Compared to the control group,the PCNA expression was decreased in CXCR4-BMSCs and BMSCs groups [(21.0±4.2) ％,(36.5±4.9) ％vs.(78.3±3.5) ％,both P＜0.05].There was a significant difference in PCNA expression between the CXCR4-BMSCsgroupandBMSCs group [(21.0±4.2)％vs.(36.5±4.9)％,P＜0.05].The neointimal area and the ratio of neointimal/medial area were decreased in CXCR4 BMSCs and BMSCs group as compared with the control group [(0.205±0.018) mm2,(0.323±0.071) mm2 vs.(0.536 ± ±0.054) mm2; (1.039±0.123),(1.660±0.404) vs.(2.460±0.328); all P＜0.05],and there were significant differences in neointimal area and the ratio of neointimal/medial area in CXCR4-BMSCs group and BMSCs group [[(0.205±0.018) mm2 vs.(0.323±0.071) mm2,(1.039±0.123)vs.(1.660±0.404),both P＜0.05].Conclusions CXCR4 gene-modified BMSCs may increase the CXCR4 expression in BMSCs.CXCR4-BMSCs transplantation is more effective than BMSCs transplantation in increasing BMSCs homing capacity,reducing the reendothelialization and vascular restenosis.
