移植途径对骨髓间质干细胞梗死心肌移植疗效的影响

The effect of implantation approach on bone marrow mesenchymal stem cells implanted into the infarct myocardium.

目的探讨自体骨髓间质干细胞（MSCs）不同移植途径对梗死心肌心肌收缩力、血管新生及胶原更新的影响。方法贵州香猪32只，随机分为对照组（C组）、冠状动脉移植组（A组）、局部注射组（T组）、局部注射＋动脉移植组（A＋T组）。抽取骨髓3ml，按照Wakitani方法培养出MSCs，经5-氮胞苷诱导后，5-溴脱氧尿苷（Brdu）标记备用。开胸结扎左冠状动脉前降支后，自体MSCs分别经结扎前降支远端灌注、局部注射、局部注射＋动脉灌注移植入自体急性心肌梗死区域，对照组以同样方法注射不含细胞的等量培养液（DMEM）。术后3、6周取标本，免疫组化检测组织血管新生、体外药物刺激离体肌条检测心肌收缩情况和血浆Ⅲ型胶原N端肽（PinNP）活性。结果MSCs移植后3周A、T、A＋T组心肌收缩恢复[（48．6±5．9）％，（42．1±6．2）％，（56．9±5．1）％]、血管新生（19．6±4．3，17．1±4．0，23．2±5．5）、血浆PⅢNP活性[（4．6±0．5）μg／L，（5．9±0．7）μg/L，（3．9±0．3）μg/L]较C组[（37．9±5．4）％，13．2±3．8，（8．7±0．8）μg／L]明显改善（P均〈0．01），且随时间推移恢复程度增加，以A＋T组为最显著（P均〈0．05）。结论局部注射＋冠状动脉灌注是较为理想的MSCs移植方式。

Objective To study the effect of different approach on myocardial contractile force, angiogenesis and collagen renovation in acute myocardial infarct site by implantation of bone marrow mesenchymal stem cells （MSCs）. Methods 32 Gui-zbou Xiang porcine were randomly divided into control group（ group C）, intra-coronary arterial injection group （group A） ,topical injection group （group T） and intra-coronary arterial ＋ topical injection group（group A ＋T）. 3 ml bone marrow was extracted. MSCs were cultured according to the methods of Wakitani＇s. After being co-cultured with 5-azacyfidine for 24 bours, these cells were labelled with bromodeoxyuridine （BrdU）. Autologus MSCs were implanted into the acute myocardial infarct site via the distal segment of the ligated left anterior descending artery （LAD） ,topical injection and both. Specimens and muscle strip were harvested in 3 and 6 weeks after MSCs implantation for microvessel count, P Ⅲ NP activity and contractile force measurement. Results The contractile force（48.6 ±5.9）% ,（42.1 ±6.2）%, （56.9 ±5.1 ）%, microvessel count（ 19.6 ±4.3,17.1 ±4.0, 23.2 ± 5.5 ） and collagen renovation （4.6 ± 0.5 ） μL, （ 5.9 ± 0.7 ） μg/L, （3.9 ± 0.3 ） μg/L in the cell implant groups was better than that in the control group（37.9 ±5.4）% ,13.2 ±3.8,（8.7 ＋0.8） μg/L,（P 〈0.01） ,especially in group A ＋ T. Conclusions Implantation of MSCs through intra-coronary arterial plus topical injection of MSCs maybe an ideal approach for infracted myocardium.