骨髓间充质干细胞移植可促进移植胰岛周围新生血管形成

李睿; 董红丽; 刘汝斌; 刘宝林

doi:10.3969/j.issn.1674-7445.2017.02.011

骨髓间充质干细胞移植可促进移植胰岛周围新生血管形成

doi: 10.3969/j.issn.1674-7445.2017.02.011

110016 沈阳，中国医科大学附属人民医院肝胆外科（李睿）；中国医科大学附属盛京医院社会服务部（董红丽），胆道、血管外科（刘宝林）；解放军第 92819 部队卫生所（刘汝斌）

基金项目:

国家自然科学基金 30972908

辽宁省科学技术基金 2013021074

详细信息

通讯作者:
刘宝林, Email: liubl55@hotmail.com

中图分类号: R587.1, R617, R-332
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出版历程
- 收稿日期: 2016-11-28
- 网络出版日期: 2021-01-19
- 刊出日期: 2017-03-15

Effect of bone mesenchymal stem cell transplantation on accelerating the vascularization surrounding transplant pancreatic islet

Department of Hepatobiliary Surgery, the People's Hospital of China Medical University, Shenyang 110016, China

More Information

Corresponding author: Liu Baolin, Email: liubl55@hotmail.com

摘要

摘要: 目的探讨胰岛移植联合骨髓间充质干细胞（MSC）移植能否促进移植胰岛周围新生血管形成。方法以非肥胖糖尿病（NOD）小鼠作为受体，将NOD小鼠随机分为4组，联合移植组（6只）、单独胰岛移植组（6只）、单独MSC移植组（6只）、假性移植组（3只）。观察各组NOD小鼠移植后血糖和存活率的变化；采用5-乙炔基-2’脱氧尿苷（EdU）及dUTP缺口末端标记（TUNEL）方法，在胰岛移植后1、2、4周检测单独胰岛移植组和联合移植组移植胰岛的增殖与凋亡情况；采用光学显微镜（光镜）直接观察、组织化学及免疫组化的方法观察并定量分析，移植术后2、4、8周单独胰岛移植组和联合移植组移植胰岛的周围新生血管密度。结果 MSC联合移植与胰岛单独移植均能明显改善移植后小鼠的血糖水平，提高NOD小鼠的存活率。MSC联合移植可促进胰岛细胞再生，减少细胞凋亡。联合移植组移植胰岛周围血管密度明显大于单独胰岛移植组。结论 MSC可以促进移植胰岛周围新生血管生成，增加移植胰岛的血供，保护移植胰岛的功能与活性。
- 1型糖尿病 /
- 胰岛移植 /
- 骨髓间充质细胞 /
- 5-乙炔基-2’脱氧尿苷（EdU） /
- dUTP缺口末端标记（TUNEL） /
- 非肥胖糖尿病（NOD）小鼠 /
- 增殖 /
- 凋亡
Abstract: Objective To investigate whether pancreatic islet transplantation in combination with bone mesenchymal stem cells (MSC) transplantation can promote the vascularization surrounding the transplant pancreatic islet. Methods The non-obese diabetic (NOD) mice were utilized as the recipients and randomly divided into pancreatic islet transplantation combined with MSC transplantation group (co-transplantation group, n=6), pancreatic islet transplantation group (n=6), MSC transplantation group (n=6) and sham transplantation group (n=3). The variation in blood glucose level and survival rate post-transplantation of NOD mice in each group was observed. The proliferation and apoptosis of the transplant pancreatic islet in the pancreatic islet transplantation group and co-transplantation group at 1, 2 and 4 weeks after pancreatic islet transplantation were analyzed by 5-ethynyl-2'-deoxyuridine (EdU) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The vascular density surrounding the transplanted pancreatic islet in the pancreatic islet transplantation group and co-transplantation group at postoperative 2, 4 and 8 weeks were observed under light microscope and quantitatively analyzed by histochemical and immunohistochemical staining. Results Both MSC combined with pancreatic islet transplantation and pancreatic islet transplantation significantly improved the blood glucose level and enhanced the survival rate of NOD mouse models after transplantation. In addition, it could accelerate the regeneration of pancreatic islet cells and decrease cell apoptosis. MSC combined with pancreatic islet transplantation significantly enhanced the vascular density surrounding the transplant pancreatic islet compared with pancreatic islet transplantation alone. Conclusions MSC transplantation can accelerate the vascularization surrounding the transplant pancreatic islet, increase the blood supply and protect the function and activity of the transplant pancreatic islet.

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图 1 移植术后不同组NOD小鼠血糖变化

Figure 1. Changes of blood glucose levels of NOD mice in different group after transplantation

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图 2 不同组NOD小鼠移植后生存曲线

Figure 2. Survival curves of NOD mice in different groups after transplantation

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图 3 移植术后两组NOD小鼠移植胰岛周围新生血管形成情况

A图为术后2周单独胰岛移植组；B图为术后2周联合移植组；C图为术后4周单独胰岛移植组；D图为术后4周联合移植组

Figure 3. Generation of new vessels in transplant islet of NOD mice in two groups after transplantation (×40)

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图 4 胰岛移植术后4周的移植胰岛的组织化学及免疫组织化学染色情况

A图为组织化学染色（HE，×100）；B图为胰岛组织insulin的表达（免疫组织化学，×100）；C图为单独胰岛移植组胰岛组织insulin的表达（免疫组织化学，×200）；D图为联合移植组胰岛组织vWF的表达（免疫组织化学，×200）

Figure 4. Histochemical and immunohistochemical staining of transplant islet at 4 weeks after transplantation

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表 1 胰岛移植不同时间后各组胰岛细胞再生与凋亡率

Table 1. Islet cell regeneration and apoptosis rate in different time after islet transplantation (x±s, %)

术后时间	单独胰岛移植组（n=6）		联合移植组（n=6）
术后时间	增殖率	凋亡率	增殖率	凋亡率
1周	0.54±0.15	1.86±0.10^b	0.63±0.04	1.43±0.13^{a, b}
2周	0.77±0.16	0.94±0.11	0.91±0.11	0.84±0.06
4周	0.89±0.05	0.87±0.11	1.01±0.10	0.76±0.07
与单独胰岛移植组比较，^aP < 0.05；与同组增殖率比较，^bP < 0.05

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