脑脉1号胶囊对移植骨髓间充质干细胞存活的影响

Effects of naomaiyihao capsule on survival of bone marrow messchymal stem cell transplantaion

目的提高骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)对靶器官植入是干细胞移植领域研究的热点。文中旨在研究调节脑中血海中药脑脉1号(NMYH)胶囊对BMSCs移植大鼠脑缺血区后存活的影响。方法采用随机法将60只SD大鼠分为模型组(无血清L-DMEM+等渗盐水)、BMSCs组(细胞悬液+等渗盐水)、NMYH组(无血清L-DMEM+NMYH)、BMSCs+NMYH组(细胞悬液+NMYH)。每组根据取材时间点又随机分为7 d、14 d、21 d组,每组每个时间点均为5只大鼠。制作大脑中动脉栓塞模型大鼠。全骨髓贴壁法分离获取第3代BMSCs,将氯甲基苯甲酰氨标记的BMSCs立体定向移植至大脑纹状体区,于移植后7、14、21 d灌注取脑,荧光显微镜下观察标记细胞在脑内存活及分布情况;免疫组织化学染色检测脑缺血边缘区血管内皮细胞标志物CD34的表达。结果第14天,BMSCs+NMYH组的荧光吸光度为1.61±0.25,较BMSCs组(1.23±0.17)显著增加(P<0.05);第21天,BMSCs+NMYH组的荧光吸光度为0.36±0.09,较BMSCs组(0.23±0.09)显著增加(P<0.05);第7、14、21天,BMSCs组、NMYH组和BMSCs+NMYH组CD34的表达较模型组均显著增加(P<0.01);第7、14、21天,BMSCs+NMYH组CD34的表达较BMSCs组均显著增加(P<0.05)。结论移植后的BMSCs能在脑组织内存活,分化,并向缺血区迁移,而NMYH可促进BMSCs移植后的存活,机制可能与其提高缺血区微血管再生,从而能增加移植细胞区域的血供有关。

Objective Nowadays ,in the field of stem cell transplantation ,improving the bone mesenchymal stem cells′ability to be implanted into target organs is still a hot topic .Objective of this study is to observe the effects of Naomaiyihao （ NMYH） capsule, a compound traditional Chinese herbal medicine invented for regulating the＂sea of blood in brain＂,and survival rate of BMSCs trans-plantaion in cerebral ischemic tissue of rat . Methods 60 rats were randomly divied into 4 group：untreated group （ serum-free L-DMEM ＋isotonic saline ） , BMSCs transplantation group （ cell suspension＋isotonic saline ） , NMYH group （ serum-free L-DMEM＋NMYH） and Combination of NMYH and BMSCs group （ cell suspen-sion＋NMYH ） .A rat of model of middle cerebral artery occlusion （ MCAO） was established .The third generation BMSCs were isolated and cultured from bone marrow by the holo-bone marrow adherence , the CM-Dil labeled BMSCs were transplanted into corpora striata with stereotactic technique , the rat′s brains were removed and sliced up＆nbsp;after 7-,14-and 21-day′transplantation .Then the survival and distribution of cells were observed under the fluorescence microscope . The expression of CD34 in the brain tissue were tested by immunohistochemical method . Results The fluorescence absorbance of combination group was 1.61 ±0.25 on day 14, increased significantly （P〈0.05）.when compared with BMSCs group （1.23 ±0.17）. The fluorescence absorbence of combination group was 0.36 ±0.09 on day 21, increased significantly （P〈0.05） when compared with BMSCs group（0.23 ±0.09）.Compared with the model group ,numbers of CD34 positive cells on day 7,14 and 21 significantly in-creased in all treated groups（P〈0.01）.Numbers of CD34 positive cells were significantly more increased（P〈0.05） in combination group than those in BMSCs group . Conclusion Survival,distribution and migration trending to boundary regions of ischemia were observed under the fluorescence microscope after transplantation .Naomaoyihao can increase blood supply around the transplanted cell region,and decrease infarct volume .Therefore it could promote BMSCs to better survive in the brain tissue .