人脐带间充质干细胞移植治疗大鼠深静脉血栓

《中国组织工程研究》 Chinese Journal of Tissue Engineering Research文章编号:2095-4344(2019)21-03392-063392www.CRTER .org·研究原著·王燕，女，1994 年生，江苏省南通市人，汉族，南通大学医学院妇产科学在读硕士，主要从事妊娠期静脉血栓栓塞症方面的研究。

通讯作者：张玉泉，博士生导师，教授，南通大学附属医院妇产科，江苏省南通市 226001文献标识码:B稿件接受：2019-02-23Wang Yan, Master candidate, Medical College of Nantong University, Nantong 226001, Jiangsu Province, ChinaCorresponding author: Zhang Yuquan, Doctoral supervisor, Professor, Department of Obstetrics and Gynecology, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China人脐带间充质干细胞移植治疗大鼠深静脉血栓王 燕1，施 沁2，张玉泉2 (1南通大学医学院，江苏省南通市 226001；2南通大学附属医院妇产科，江苏省南通市 226001)DOI:10.3969/j.issn.2095-4344.1745 ORCID: 0000-0002-2939-0036(王燕)文章快速阅读：文题释义：脐带间充质干细胞：人脐带间充质干细胞是来源于发育早期中胚层和外胚层的一类多能干细胞，具有自我增殖及多向分化、免疫调节、归巢、旁分泌、组织修复及促血管新生等特性，严格意义上其来源分4种，即华通氏胶、脐血、脐血管及脐血管周围细胞，实验采用脐带华通氏胶源性间充质干细胞。

血栓溶解、机化及再通：在血栓形成后，纤维蛋白溶解酶激活以及血栓内白细胞释放溶蛋白酶，较小的血栓可以被完全溶解吸收，若血栓较大或纤维蛋白溶解酶活性不足，存留的血栓则发生机化及再通。

内皮细胞、成纤维细胞和炎性细胞从血管壁长入血栓，由肉芽组织逐渐取代血栓的过程，为血栓机化。

过程中，血管内皮细胞长入并被覆于血栓内裂隙表面，形成新的血管，相互吻合成新的管腔，血流部分恢复，这一过程称为再通。

摘要背景：人脐带间充质干细胞具有归巢至缺血部位并在局部促进血管新生的能力，而新血管生成对深静脉血栓的机化及再通至关重要，但是将人脐带间充质干细胞用于血栓的研究尚未见相关报道。

目的：探讨人脐带间充质干细胞移植治疗对深静脉血栓溶解、机化及再通的影响。

方法：用“狭窄法”建立下腔静脉血栓模型大鼠，制备P3代人脐带间充质干细胞悬液，细胞浓度2×109 L -1，将建模后大鼠随机分为4组。

对照组：建模后第1，2，3天尾静脉注射PBS 1 mL ；实验组1：术后第1天尾静脉注射细胞悬液1 mL ，第2，3天注射PBS 1 mL ；实验组2：术后第1，2天注射细胞悬液1 mL ，第3天注射PBS 1 mL ；实验组3：术后第1，2，3天注射细胞悬液1 mL 。

实验方案经南通大学动物实验伦理委员会批准，批准号为20180315-001，批准时间2018-04-01。

于术后第7天取出下腔静脉内血栓，通过苏木精-伊红染色及免疫组织化学染色观察比较各组血栓内新生血管数及溶解率。

结果与结论：①与对照组相比，实验组2，3血栓内血管密度显著增高(P < 0.01)；②所有实验组的溶解率较对照组增高，但差异无显著性意义(P > 0.05)；③结果表明，一定剂量人脐带间充质干细胞移植能显著增加血栓内的小血管数量，有助于血栓的机化及再通。

关键词：深静脉血栓；机化；再通；人脐带间充质干细胞；血管新生；血管密度；溶解率；下腔静脉血栓模型；大鼠；干细胞中图分类号：R453；R394.2 基金资助：江苏省医学创新团队[苏卫科教(2017)1号]，项目负责人：张玉泉Treating deep vein thrombosis with human umbilical cord mesenchymal stem cell transplantationWang Yan 1, Shi Qin 2, Zhang Yuquan 2 (1Medical College of Nantong University, Nantong 226001, JiangsuProvince, China; 2Department of Obstetrics and Gynecology, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China)Wang Y, Shi Q, Zhang YQ. Treating deep vein thrombosis with human umbilical cord mesenchymal stem cell transplantation.Zhongguo Zuzhi Gongcheng Yanjiu. 2019;23(21):3392-3397. DOI:10.3969/j.issn.2095-4344.1745P .O. Box 10002, Shenyang 110180 3393AbstractBACKGROUND: Human umbilical cord mesenchymal stem cells can home to ischemic sites and form functional blood vessels locally. Neovascularization is critical for the organization and recanalization of deep vein thrombosis. However, transplantation of human umbilical cord mesenchymal stem cells for the treatment of thrombosis has not been reported.OBJECTIVE: To investigate the dissolution, organization and recanalization of deep vein thrombosis after the transplantation of human umbilical cord mesenchymal stem cells.METHODS: Deep vein thrombosis model was established by “narrow method” in rats. Suspension of the third generation human umbilical cord mesenchymal stem cells at a density of 2×109 cells/L was prepared. The model rats were randomly divided into four groups. In control group, 1 mL of PBS was injected into the tail vein on days 1, 2, and 3 after modeling. In experimental group 1, 1 mL of cell suspension was injected into the tail vein on day 1 after modeling, and 1 mL of PBS was injected on days 2 and 3. In experimental group 2, 1 mL of cell suspension was injected on days 1 and 2 after modeling, and 1 mL of PBS was injected on day 3. In experimental group 3, 1 mL of cell suspension was injected on days 1, 2, and 3 after modeling. The experimental protocol was approved by the Animal Experimental Ethics Committee of Nantong University with the approval No. 20180315-001 on April 1, 2018. Samples from deep vein thrombosis rats were taken on day 7 after modeling. Number of new blood vessels and dissolution rate in each group were detected by hematoxylin-eosin staining and immunohistochemical staining.RESULTS AND CONCLUSION: Compared with the control group, the capillary density of the experimental groups 2 and 3 was significantly increased (P < 0.01). The dissolution rate of all the experimental groups was higher than that of the control group, but the difference was not statistically significant (P > 0.05). All these results reveal that a certain amount of human umbilical cord mesenchymal stem cells transplanted can increase the number of microvessels in the thrombosis, and accelerate organization and recanalization of the thrombosis.Key words: deep vein thrombosis; organization; recanalization; human umbilical cord mesenchymal stem cells; angiogenesis; vascular density; dissolution rate; inferior vena cava thrombosis model; rat; stem cellsFunding: Jiangsu Province Medical Innovation Team, No. Su Wei Ke Jiao(2017)1 (to ZYQ)0 引言 Introduction深静脉血栓是一类具有较高发生率和病死率的血管性疾病。