柴胡龙骨牡蛎汤对心肌梗死大鼠骨髓c-kit^+干细胞动员的影响

Influence of Chaihu Longgu Muli Tang on mobilization of bone marrow c-kit^+ stem cells in rats with myocardial infarction

目的探讨柴胡龙骨牡蛎汤对心肌梗死后大鼠骨髓c-kit^+干细胞动员的影响。方法将雄性SD大鼠随机分为假手术组、模型组、柴胡龙骨牡蛎汤组,各个组分设3、7和14 d 3个亚组。柴胡龙骨牡蛎汤组给予柴胡龙骨牡蛎汤灌胃,其余各组给予蒸馏水灌胃。采用心脏超声评价心功能,HE染色评价心肌病理变化,Masson染色观察心肌纤维化程度,流式细胞术检测骨髓和外周血ckit^+阳性细胞数,免疫组化法检测心肌梗死边缘区c-kit^+阳性细胞数。结果模型组大鼠心功能随心肌梗死时间延长逐渐下降,与假手术组相比,各个时间点差异均具有统计学意义(P<0.05)。柴胡龙骨牡蛎汤组大鼠的心功能趋于稳定状态且优于模型组,与模型组相比,左室射血分数、左室短轴缩短率7 d和14 d差异具有统计学意义(P<0.05)。病理染色显示假手术组心肌细胞排列整齐,无胶原纤维,模型组出现心肌细胞坏死和纤维化,且随着心梗时间延长逐渐加剧;柴胡龙骨牡蛎汤组心肌细胞坏死和纤维化程度均较模型组减轻。模型组和柴胡龙骨牡蛎汤组骨髓和外周血ckit^+阳性细胞数均在心梗后7 d达到高峰,与假手术组相比差异具有统计学意义(P<0.05);柴胡龙骨牡蛎汤组高于模型组,7 d和14 d组间相比差异具有统计学意义(P<0.05)。模型组心肌梗死边缘区c-kit^+阳性细胞数高于假手术组,7 d和14 d组间相比差异具有统计学意义(P<0.05),柴胡龙骨牡蛎汤组心肌梗死边缘区c-kit^+阳性细胞高于模型组,7 d和14 d组间相比差异具有统计学意义(P<0.05)。结论柴胡龙骨牡蛎汤促进心肌梗死后骨髓c-kit^+干细胞的动员,这可能是其改善心功能的机制之一。

Objective To investigate the influence of Chaihu Longgu Muli Tang（ Chinese Thorowax Root,Bone Fossil of Big Mammals and Oyster Shell Decoction,CLMT） on the mobilization of bone marrow c-kit~＋ stem cells in rats after myocardial infarction（ MI）. Methods Male SD rats were randomly divided into sham-operation group,model group and CLMT group,and each group was divided again into 3-d sub group,7-d subgroup and 14-d subgroup. The CLMT group was intragastrically given CLMT,and other groups were orally given distilled water. The heart function was reviewed by using echocardiogram,changes of myocardial pathology were reviewed after HE staining,and severity of myocardial fibrosis was observed by after Masson staining. The number of c-kit~＋ positive cells in bone marrow and peripheral blood was detected by using flow cytometry（ FCM）,and number of c-kit~＋ positive cells in MI border zone was detected by using immunohistochemistry technique. Results The heart function declined gradually along with MI time extension in model group,and difference in heart function had statistical significance compared with sham-operation group at all time points（ P〈0. 05）. The heart function trended to stable in CLMT group and was better than that in model group,and difference in LVEF and LVFS had statistical significance compared with model group at time points of 7 d and 14 d（ P〈0. 05）.Pathological staining showed that myocardial cells were in alignment without collagen fibers in shamoperation group,necrosis and fibrosis of myocardial cells in model group that aggravated gradually along with MI time extension. The severity of myocardial necrosis and fibrosis was milder in CLMT group than that in model group. The number of c-kit~＋ positive cells in bone marrow and peripheral blood reached peak in model group and CLMT group 7 d after MI,and difference number of c-kit~＋ positive cells had statistical significance compared with sham-operation group,and was higher in CLMT group than that in model group. The difference in number of c-kit~＋ positive cells had statistical significance between 7-d subgroup and 14-d subgroup（ P〈0. 05）. The number of c-kit~＋ positive cells in MI border zone was higher in model group than that in sham-operation group,and difference in number of c-kit~＋ positive cells had statistical significance between 7-d subgroup and 14-d subgroup（ P〈0. 05）. The number of c-kit~＋ positive cells in MI border zone was higher in CLMT group than that in model group,and difference in number of c-kit~＋ positive cells MI border zone had statistical significance between 7-d subgroup and 14-d subgroup（ P〈0. 05）. Conclusion CLMT can improve the mobilization of bone marrow c-kit~＋ stem cells after MI and heart function.