CAJ | 학술논문

目的观察移植过表达缝隙连接蛋白43(Cx43)的骨髓间充质干细胞(BMSC)对心肌梗死后心力衰竭大鼠的治疗作用.方法将60只SD大鼠分为4组,每组15只.假手术组;DMEM/F12组:注射DMEM/F12培养基;EGFP(增强的绿荧光蛋白)组:移植转染EGFP 基因的BMSC;Cx43组:移植转染Cx43基因的BMSC.采用结扎前降支的方法,建立心肌梗死模型,于梗死后30 min,行细胞移植.移植后4周,超声心动图检测心脏功能并取心脏组织.Masson染色检测梗死面积及胶原纤维含量.激光共聚焦显微镜观察EGFP基因及Cx43的表达.RT-PCR检测移植区心肌组织Cx43 mRNA含量.结果 EGFP组和Cx43组心功能明显改善[左室射血分数分别为(76.0±6.9)%、(82.9±5.0)%,P＜0.05],心肌梗死面积缩小,胶原纤维含量降低[分别为(13.6±4.7)%、(15.9±3.2)%,P＞0.05],Cx43组心脏重构指标改善更明显.激光共聚焦显示EGFP组和Cx43组均有BMSC存活,并与宿主心肌组织间形成缝隙连接.DMEM/F12 组Cx43 mRNA水平较假手术组显著降低(0.18±0.05比0.78±0.14,P<0.01),EGFP组Cx43 mRNA的含量与DMEM/F12 组相比差异无统计学意义(0.20±0.09 比 0.18±0.05,P>0.05).Cx43组Cx43 mRNA水平较DMEM/F12 组及EGFP组明显增加(0.39±0.14 比 0.18±0.05,P<0.01;0.39±0.14比0.20±0.09,P<0.05).结论过表达Cx43 的BMSC移植改善心力衰竭的效果更为明显,可能与BMSC改善了移植区缝隙连接表达有关.
목적 관찰이식과표체봉극련접단백43(Cx43)적골수간충질간세포(BMSC)대심기경사후심력쇠갈대서적치료작용.방법 장60지SD대서분위4조,매조15지.가수술조;DMEM/F12조:주사DMEM/F12배양기;EGFP(증강적록형광단백)조:이식전염EGFP 기인적BMSC;Cx43조:이식전염Cx43기인적BMSC.채용결찰전강지적방법,건립심기경사모형,우경사후30 min,행세포이식.이식후4주,초성심동도검측심장공능병취심장조직.Masson염색검측경사면적급효원섬유함량.격광공취초현미경관찰EGFP기인급Cx43적표체.RT-PCR검측이식구심기조직Cx43 mRNA함량.결과 EGFP조화Cx43조심공능명현개선[좌실사혈분수분별위(76.0±6.9)%、(82.9±5.0)%,P＜0.05],심기경사면적축소,효원섬유함량강저[분별위(13.6±4.7)%、(15.9±3.2)%,P＞0.05],Cx43조심장중구지표개선경명현.격광공취초현시EGFP조화Cx43조균유BMSC존활,병여숙주심기조직간형성봉극련접.DMEM/F12 조Cx43 mRNA수평교가수술조현저강저(0.18±0.05비0.78±0.14,P<0.01),EGFP조Cx43 mRNA적함량여DMEM/F12 조상비차이무통계학의의(0.20±0.09 비 0.18±0.05,P>0.05).Cx43조Cx43 mRNA수평교DMEM/F12 조급EGFP조명현증가(0.39±0.14 비 0.18±0.05,P<0.01;0.39±0.14비0.20±0.09,P<0.05).결론 과표체Cx43 적BMSC이식개선심력쇠갈적효과경위명현,가능여BMSC개선료이식구봉극련접표체유관.
Objective To explore the therapeutic effect of transplantation of bone marrow mesenchymal stem cells (BMSCs) over-expressing Cx43 on heart failure in post-infarction rats. Methods Sixty SD rats were randomly divided equally into 4 groups: sham group , DMEM/F12 group injected with DMEM/F12, EGFP group with transplanted EGFP transfected BMSCs and Cx43 group with transplanted Cx43 transfected BMSCs. Myocardial infarction model was established by ligating anterior descending branch and the cells were transplanted after 30 minutes. At Week 4 post-infarction, the heart functions of rats were evaluated by echocardiography. After the rats were sacrificed, their tissue samples were collected. The areas of myocardial infarction and the levels of collagen fiber content were measured. And the expressions of EGFR and Cx43 were observed under laser confocal microscopy. The level of Cx43 mRNA was measured by reverse transcription-polymerase chain reaction (RT-PCR). Results As compared with the DMEM/F12 group, cardiac function was improved significantly, myocardial infarct area shrunk and collagen fiber content decreased significantly in the EGFP and group in Cx43 groups the . Survival of BMSCs and the formation of gap junction between BMSCs and the host myocardium could be observed under laser confocal microscopy both in EGFP group and Cx43 groups. And the post-infarction, expression of Cx43 mRNA in myocardial tissue decreased significantly in the group DMEM/F12, when compared with sham group(0.18±0.05 vs 0.78±0.14, P<0.01). There was no significant difference on expression of Cx43 mRNA between DMEM/F12 group and EGFP group (0.18±0.05 vs 0.20±0.09, P>0.05). The lever of Cx43 mRNA was higher in group Cx43 than in group DMEM/F12 and group EGFP(0.39±0.14 vs 0.18±0.05, P<0.01; 0.39±0.14 vs 0.20±0.09, P<0.05). Conclusion Transplantation of BMSCs attenuates ventricular remodeling and improves cardiac functions. It may result from the over-expression of Cx43 gene through its effects of improving gap junction remodeling and increasing electro-mechanical coupling between myocardial cells in peri-infarct area.