新生猪肾包膜下胰岛移植动物模型构建的初步研究

张国强; 李世朋; 李煜; 王峰; 唐华侨; 张志; 王有财; 刘英俊; 王刚成

doi:10.3969/j.issn.1674-7445.2021.06.013

新生猪肾包膜下胰岛移植动物模型构建的初步研究

doi: 10.3969/j.issn.1674-7445.2021.06.013

450000 郑州大学附属肿瘤医院河南省肿瘤医院普外科（张国强、张志、王有财、刘英俊、王刚成）；首都医科大学附属北京友谊医院肝脏移植中心（李世朋）；南昌大学第一附属医院泌尿外科（李煜）；青岛大学附属医院器官移植中心（王峰）；四川农业大学动物医学院（唐华侨）

基金项目:

河南省科技攻关重点项目 202102310109

河南省卫生健康委省部共建重点项目 SBGJ202002018

河南省卫生健康委省部共建重点项目 SB201901109

详细信息

作者简介:
张国强，博士，主治医师，研究方向为猪器官异种移植，Email：171891625@qq.com

通讯作者:
王刚成，主任医师、教授，硕士研究生导师，Email：13937152656@163.com

中图分类号: R617, R587.1
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出版历程
- 收稿日期: 2021-07-31
- 刊出日期: 2021-11-15

Preliminary study on the establishment of neonatal pig models of islet transplantation under the renal capsule

Department of General Surgery, Affiliated Cancer Hospital of Zhengzhou University, Henan Provincial Cancer Hospital, Zhengzhou 450000, China

More Information

Corresponding author: Wang Gangcheng, Email: 13937152656@163.com

摘要

摘要: 目的构建新生猪肾包膜下胰岛移植模型，探讨其可行性和潜在应用价值。方法选取9只野生型同胎新生健康杜洛克猪，其中1只为对照（p6307），6只为胰岛移植供体，2只为胰岛移植受体（p6210、p6207）。新生猪胰岛分离、体外培养分化后，行猪肾包膜下胰岛移植术，术后给予他克莫司（Tac）联合西罗莫司免疫抑制治疗。术后监测移植受体的体质量、血糖、血清肌酐水平，术后4周和8周分别处死受体新生猪p6210和p6207、对照新生猪，获取猪肾包膜下胰岛移植物进行病理学染色和胰岛素免疫荧光染色。结果猪肾包膜下胰岛移植术后，受体新生猪的体质量增长相对于对照新生猪明显减慢，并间断性伴有纳差、腹泻等症状；而受体新生猪的血糖、血清肌酐水平相较于术前及对照新生猪无明显变化。受体新生猪p6210肾包膜下可见明显的胰岛团块，病理学染色和胰岛素免疫荧光染色证实胰岛团块具有分泌胰岛素的功能，而受体新生猪p6207肾包膜下未见明显的胰岛团块，病理学染色未见明显胰岛团块，考虑该受体新生猪发生胰岛移植排斥反应导致胰岛定植失败。结论新生猪肾包膜下胰岛移植是可行的，为下一步构建用于异种移植治疗糖尿病终末期肾病的猪胰岛-肾复合供体器官提供前期铺垫。
- 杜洛克猪 /
- 胰岛移植 /
- 肾包膜 /
- 异种移植 /
- 胰岛素 /
- 糖尿病 /
- 立即经血液介导的炎症反应（IBMIR）
Abstract: Objective To explore the feasibility and potential application value of establishing the neonatal pig models of islet transplantation under the renal capsule. Methods Nine wild-type neonatal Duroc pigs were selected, including 1 animal as the control (p6307), 6 as islet transplant donors and 2 as islet transplant recipients (p6210, p6207). After islet isolation and differentiation in vitro, islet transplantation under the renal capsule of the pig was performed. Immunosuppressive therapy of tacrolimus (Tac) combined with sirolimus was given after operation. Postoperative body weight, blood glucose and serum creatinine levels of the recipients were monitored. The p6210 recipient neonatal pig was sacrificed at postoperative 4 weeks, while the p6207 recipient and the control neonatal pig were sacrificed at postoperative 8 weeks. The islet grafts under the renal capsule were collected for pathological staining and insulin immunofluorescent staining. Results After islet transplantation under the renal capsule of the pigs, the growth rate of body weight of the recipients was significantly slower than that of the control neonatal pig, accompanied with intermittent symptoms, such as anorexia and diarrhea, etc. However, the blood glucose and serum creatinine levels of the recipients did not significantly differ from preoperative levels and those of the control neonatal pig. Evident islet mass was observed under the renal capsule of the p6210 recipient. Pathological staining and insulin immunofluorescent staining confirmed that the islet mass had the function of secreting insulin, whereas no obvious islet mass could be seen under the renal capsule of the p6207 recipient. Pathological staining detected no evident islet mass, suggesting the possibility of islet transplantation failure caused by rejection in the p6207 recipient. Conclusions The establishment of neonatal pig models of islet transplantation under the renal capsule is a feasible technique, which provides preliminary evidence for the establishment of composite islet-kidney donor graft in pig models for xenotransplantation in the treatment of end-stage diabetic nephropathy.
- Duroc pig /
- Islet transplantation /
- Renal capsule /
- Xenotransplantation /
- Insulin /
- Diabetes mellitus /
- Instant blood-mediated inflammatory response (IBMIR)

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图 1 猪肾包膜下胰岛移植手术步骤

注：A、B图为游离肾脂肪囊、建立肾包膜下移植空间；C~F图为肾包膜下注入胰岛细胞，移植术后缝闭肾包膜。

Figure 1. Surgical steps of islet transplantation under the renal capsule of pigs

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图 2 胰岛移植术后受体的体质量、血糖及血清肌酐水平变化

注：A图为体质量变化；B图为血糖水平变化；C图为血清肌酐水平变化。

Figure 2. Changes of body weight, blood glucose and serum creatinine levels of recipients after islet transplantation

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图 3 猪肾包膜下胰岛移植物的大体标本

注：A图示受体新生猪p6210肾包膜下可见胰岛团块（胰岛移植术后4周）；B图示受体新生猪p6207肾包膜下未见明显胰岛团块（胰岛移植术后8周）。

Figure 3. Gross specimen of islet graft under the renal capsule of pigs

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图 4 胰岛移植物病理学染色及免疫荧光染色结果

注：A图为受体新生猪p6210肾包膜下胰岛移植物病理学染色结果（苏木素-伊红，×40）；B图为受体新生猪p6207肾包膜下胰岛移植物病理学染色结果（苏木素-伊红，×100）；C图为受体新生猪p6210肾包膜下胰岛移植物胰岛素染色结果，红色为胰岛素阳性（免疫荧光，×200）。

Figure 4. Results of pathological staining and immunofluorescence staining of islet grafts

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