骨髓基质干细胞及其来源的产胰岛素细胞移植对受体胰岛和新生血管的影响

Objective To investigate the proliferative impact of transplantation of bone marrow mesenchymal stem cells (MSCs) and insulin producing ceils (IPCs) from them on recipients' remnant islets and neovascularization around. Methods MSCs were induced to IPCs in vitro. The rats with type Ⅰ diabetic meilitus were randomly divided into control group, MSCs transplant group, and IPCs transplant group. When the blood glucose level of recipient rats was decreased to 10 mmol/L, the right kidney and pancreas were removed from each group simultaneously. All the thin tissue slices were estimated by immu-nohistochemistry staining of PCNA antibody,insulin antibody and CD31 anbibody. Results The passaged MSCs differentiated into IPCs. In the grafts, insulin-positive and CD31 -positive cells were observed in the right kidney. In control group, few proliferative cells located in the atrophic islets. In the transplant groups, a lot of proliferative islet cells were obsereved, around which few CD31 -positive ceils existed;Pancreatic proliferative islet cell rate in MSCs and IPCs transplant group was ( 20.84±3.48 ) % and ( 18.43± 2.84% ) ( P > 0.05 ) respectively. Conclusion Obviously, MSCs and IPCs promoted recipient neovascu-larization surrounding the islets, which enhanced the proliferation of remnant islet cells. MSCs differentiated to IPCs and vascular endothelial cells in the recipient transplant location.     

骨髓基质干细胞及其来源的产胰岛素细胞移植对受体胰岛和新生血管的影响

Objective To investigate the proliferative impact of transplantation of bone marrow mesenchymal stem cells (MSCs) and insulin producing ceils (IPCs) from them on recipients' remnant islets and neovascularization around. Methods MSCs were induced to IPCs in vitro. The rats with type Ⅰ diabetic meilitus were randomly divided into control group, MSCs transplant group, and IPCs transplant group. When the blood glucose level of recipient rats was decreased to 10 mmol/L, the right kidney and pancreas were removed from each group simultaneously. All the thin tissue slices were estimated by immu-nohistochemistry staining of PCNA antibody,insulin antibody and CD31 anbibody. Results The passaged MSCs differentiated into IPCs. In the grafts, insulin-positive and CD31 -positive cells were observed in the right kidney. In control group, few proliferative cells located in the atrophic islets. In the transplant groups, a lot of proliferative islet cells were obsereved, around which few CD31 -positive ceils existed;Pancreatic proliferative islet cell rate in MSCs and IPCs transplant group was ( 20.84±3.48 ) % and ( 18.43± 2.84% ) ( P > 0.05 ) respectively. Conclusion Obviously, MSCs and IPCs promoted recipient neovascu-larization surrounding the islets, which enhanced the proliferation of remnant islet cells. MSCs differentiated to IPCs and vascular endothelial cells in the recipient transplant location.     

骨髓基质干细胞及其来源的产胰岛素细胞移植对受体胰岛和新生血管的影响

Objective To investigate the proliferative impact of transplantation of bone marrow mesenchymal stem cells (MSCs) and insulin producing ceils (IPCs) from them on recipients' remnant islets and neovascularization around. Methods MSCs were induced to IPCs in vitro. The rats with type Ⅰ diabetic meilitus were randomly divided into control group, MSCs transplant group, and IPCs transplant group. When the blood glucose level of recipient rats was decreased to 10 mmol/L, the right kidney and pancreas were removed from each group simultaneously. All the thin tissue slices were estimated by immu-nohistochemistry staining of PCNA antibody,insulin antibody and CD31 anbibody. Results The passaged MSCs differentiated into IPCs. In the grafts, insulin-positive and CD31 -positive cells were observed in the right kidney. In control group, few proliferative cells located in the atrophic islets. In the transplant groups, a lot of proliferative islet cells were obsereved, around which few CD31 -positive ceils existed;Pancreatic proliferative islet cell rate in MSCs and IPCs transplant group was ( 20.84±3.48 ) % and ( 18.43± 2.84% ) ( P > 0.05 ) respectively. Conclusion Obviously, MSCs and IPCs promoted recipient neovascu-larization surrounding the islets, which enhanced the proliferation of remnant islet cells. MSCs differentiated to IPCs and vascular endothelial cells in the recipient transplant location.     

骨髓基质干细胞及其来源的产胰岛素细胞移植对受体胰岛和新生血管的影响

Objective To investigate the proliferative impact of transplantation of bone marrow mesenchymal stem cells (MSCs) and insulin producing ceils (IPCs) from them on recipients' remnant islets and neovascularization around. Methods MSCs were induced to IPCs in vitro. The rats with type Ⅰ diabetic meilitus were randomly divided into control group, MSCs transplant group, and IPCs transplant group. When the blood glucose level of recipient rats was decreased to 10 mmol/L, the right kidney and pancreas were removed from each group simultaneously. All the thin tissue slices were estimated by immu-nohistochemistry staining of PCNA antibody,insulin antibody and CD31 anbibody. Results The passaged MSCs differentiated into IPCs. In the grafts, insulin-positive and CD31 -positive cells were observed in the right kidney. In control group, few proliferative cells located in the atrophic islets. In the transplant groups, a lot of proliferative islet cells were obsereved, around which few CD31 -positive ceils existed;Pancreatic proliferative islet cell rate in MSCs and IPCs transplant group was ( 20.84±3.48 ) % and ( 18.43± 2.84% ) ( P > 0.05 ) respectively. Conclusion Obviously, MSCs and IPCs promoted recipient neovascu-larization surrounding the islets, which enhanced the proliferation of remnant islet cells. MSCs differentiated to IPCs and vascular endothelial cells in the recipient transplant location.     

骨髓基质干细胞及其来源的产胰岛素细胞移植对受体胰岛和新生血管的影响

Objective To investigate the proliferative impact of transplantation of bone marrow mesenchymal stem cells (MSCs) and insulin producing ceils (IPCs) from them on recipients' remnant islets and neovascularization around. Methods MSCs were induced to IPCs in vitro. The rats with type Ⅰ diabetic meilitus were randomly divided into control group, MSCs transplant group, and IPCs transplant group. When the blood glucose level of recipient rats was decreased to 10 mmol/L, the right kidney and pancreas were removed from each group simultaneously. All the thin tissue slices were estimated by immu-nohistochemistry staining of PCNA antibody,insulin antibody and CD31 anbibody. Results The passaged MSCs differentiated into IPCs. In the grafts, insulin-positive and CD31 -positive cells were observed in the right kidney. In control group, few proliferative cells located in the atrophic islets. In the transplant groups, a lot of proliferative islet cells were obsereved, around which few CD31 -positive ceils existed;Pancreatic proliferative islet cell rate in MSCs and IPCs transplant group was ( 20.84±3.48 ) % and ( 18.43± 2.84% ) ( P > 0.05 ) respectively. Conclusion Obviously, MSCs and IPCs promoted recipient neovascu-larization surrounding the islets, which enhanced the proliferation of remnant islet cells. MSCs differentiated to IPCs and vascular endothelial cells in the recipient transplant location.     

骨髓基质干细胞及其来源的产胰岛素细胞移植对受体胰岛和新生血管的影响

Objective To investigate the proliferative impact of transplantation of bone marrow mesenchymal stem cells (MSCs) and insulin producing ceils (IPCs) from them on recipients' remnant islets and neovascularization around. Methods MSCs were induced to IPCs in vitro. The rats with type Ⅰ diabetic meilitus were randomly divided into control group, MSCs transplant group, and IPCs transplant group. When the blood glucose level of recipient rats was decreased to 10 mmol/L, the right kidney and pancreas were removed from each group simultaneously. All the thin tissue slices were estimated by immu-nohistochemistry staining of PCNA antibody,insulin antibody and CD31 anbibody. Results The passaged MSCs differentiated into IPCs. In the grafts, insulin-positive and CD31 -positive cells were observed in the right kidney. In control group, few proliferative cells located in the atrophic islets. In the transplant groups, a lot of proliferative islet cells were obsereved, around which few CD31 -positive ceils existed;Pancreatic proliferative islet cell rate in MSCs and IPCs transplant group was ( 20.84±3.48 ) % and ( 18.43± 2.84% ) ( P > 0.05 ) respectively. Conclusion Obviously, MSCs and IPCs promoted recipient neovascu-larization surrounding the islets, which enhanced the proliferation of remnant islet cells. MSCs differentiated to IPCs and vascular endothelial cells in the recipient transplant location.     

骨髓基质干细胞及其来源的产胰岛素细胞移植对受体胰岛和新生血管的影响

Objective To investigate the proliferative impact of transplantation of bone marrow mesenchymal stem cells (MSCs) and insulin producing ceils (IPCs) from them on recipients' remnant islets and neovascularization around. Methods MSCs were induced to IPCs in vitro. The rats with type Ⅰ diabetic meilitus were randomly divided into control group, MSCs transplant group, and IPCs transplant group. When the blood glucose level of recipient rats was decreased to 10 mmol/L, the right kidney and pancreas were removed from each group simultaneously. All the thin tissue slices were estimated by immu-nohistochemistry staining of PCNA antibody,insulin antibody and CD31 anbibody. Results The passaged MSCs differentiated into IPCs. In the grafts, insulin-positive and CD31 -positive cells were observed in the right kidney. In control group, few proliferative cells located in the atrophic islets. In the transplant groups, a lot of proliferative islet cells were obsereved, around which few CD31 -positive ceils existed;Pancreatic proliferative islet cell rate in MSCs and IPCs transplant group was ( 20.84±3.48 ) % and ( 18.43± 2.84% ) ( P > 0.05 ) respectively. Conclusion Obviously, MSCs and IPCs promoted recipient neovascu-larization surrounding the islets, which enhanced the proliferation of remnant islet cells. MSCs differentiated to IPCs and vascular endothelial cells in the recipient transplant location.     

骨髓基质干细胞及其来源的产胰岛素细胞移植对受体胰岛和新生血管的影响

Objective To investigate the proliferative impact of transplantation of bone marrow mesenchymal stem cells (MSCs) and insulin producing ceils (IPCs) from them on recipients' remnant islets and neovascularization around. Methods MSCs were induced to IPCs in vitro. The rats with type Ⅰ diabetic meilitus were randomly divided into control group, MSCs transplant group, and IPCs transplant group. When the blood glucose level of recipient rats was decreased to 10 mmol/L, the right kidney and pancreas were removed from each group simultaneously. All the thin tissue slices were estimated by immu-nohistochemistry staining of PCNA antibody,insulin antibody and CD31 anbibody. Results The passaged MSCs differentiated into IPCs. In the grafts, insulin-positive and CD31 -positive cells were observed in the right kidney. In control group, few proliferative cells located in the atrophic islets. In the transplant groups, a lot of proliferative islet cells were obsereved, around which few CD31 -positive ceils existed;Pancreatic proliferative islet cell rate in MSCs and IPCs transplant group was ( 20.84±3.48 ) % and ( 18.43± 2.84% ) ( P > 0.05 ) respectively. Conclusion Obviously, MSCs and IPCs promoted recipient neovascu-larization surrounding the islets, which enhanced the proliferation of remnant islet cells. MSCs differentiated to IPCs and vascular endothelial cells in the recipient transplant location.     

骨髓基质干细胞及其来源的产胰岛素细胞移植对受体胰岛和新生血管的影响

Objective To investigate the proliferative impact of transplantation of bone marrow mesenchymal stem cells (MSCs) and insulin producing ceils (IPCs) from them on recipients' remnant islets and neovascularization around. Methods MSCs were induced to IPCs in vitro. The rats with type Ⅰ diabetic meilitus were randomly divided into control group, MSCs transplant group, and IPCs transplant group. When the blood glucose level of recipient rats was decreased to 10 mmol/L, the right kidney and pancreas were removed from each group simultaneously. All the thin tissue slices were estimated by immu-nohistochemistry staining of PCNA antibody,insulin antibody and CD31 anbibody. Results The passaged MSCs differentiated into IPCs. In the grafts, insulin-positive and CD31 -positive cells were observed in the right kidney. In control group, few proliferative cells located in the atrophic islets. In the transplant groups, a lot of proliferative islet cells were obsereved, around which few CD31 -positive ceils existed;Pancreatic proliferative islet cell rate in MSCs and IPCs transplant group was ( 20.84±3.48 ) % and ( 18.43± 2.84% ) ( P > 0.05 ) respectively. Conclusion Obviously, MSCs and IPCs promoted recipient neovascu-larization surrounding the islets, which enhanced the proliferation of remnant islet cells. MSCs differentiated to IPCs and vascular endothelial cells in the recipient transplant location.     

骨髓基质干细胞及其来源的产胰岛素细胞移植对受体胰岛和新生血管的影响

Objective To investigate the proliferative impact of transplantation of bone marrow mesenchymal stem cells (MSCs) and insulin producing ceils (IPCs) from them on recipients' remnant islets and neovascularization around. Methods MSCs were induced to IPCs in vitro. The rats with type Ⅰ diabetic meilitus were randomly divided into control group, MSCs transplant group, and IPCs transplant group. When the blood glucose level of recipient rats was decreased to 10 mmol/L, the right kidney and pancreas were removed from each group simultaneously. All the thin tissue slices were estimated by immu-nohistochemistry staining of PCNA antibody,insulin antibody and CD31 anbibody. Results The passaged MSCs differentiated into IPCs. In the grafts, insulin-positive and CD31 -positive cells were observed in the right kidney. In control group, few proliferative cells located in the atrophic islets. In the transplant groups, a lot of proliferative islet cells were obsereved, around which few CD31 -positive ceils existed;Pancreatic proliferative islet cell rate in MSCs and IPCs transplant group was ( 20.84±3.48 ) % and ( 18.43± 2.84% ) ( P > 0.05 ) respectively. Conclusion Obviously, MSCs and IPCs promoted recipient neovascu-larization surrounding the islets, which enhanced the proliferation of remnant islet cells. MSCs differentiated to IPCs and vascular endothelial cells in the recipient transplant location.     

骨髓基质干细胞及其来源的产胰岛素细胞移植对受体胰岛和新生血管的影响

目的 观察骨髓基质干细胞(MSCs)及其来源的产胰岛素细胞(IPCs)移植对受体残余胰岛和其周围新生血管增殖的影响.方法 对大鼠MSCs进行体外诱导成为IPCs.对1型糖尿病大鼠随机分为胰岛素控制血糖组(对照组)、MSCs移植组及IPCs移植组3个治疗组(每组10只).至血糖开始下降至10mmoL/L,同期移除3组大鼠右肾及胰腺,分别进行PCNA、胰岛素和CD31抗体染色,观察3组大鼠肾脏和胰腺的胰岛素及血管内皮细胞表达情况.结果 MSCs诱导生成IPCs.移植物:移植侧的右肾均可见到较多的胰岛素和CD31阳性细胞.胰腺组织:(1)对照组:胰岛萎缩.(2)细胞移植组:残余胰岛增殖率:(20.84±3.48)%和(18.43±2.84)%(P>0.05),胰岛周围均有少量CD31阳性细胞.结论 MSCs及其来源的IPCs明显促进了受体残余胰岛周围新生血管的生成,进而使残余胰岛得到增殖,而MSCs在体内也可分化成IPCs和血管内皮细胞.     

骨髓间充质干细胞移植对大鼠脊髓损伤后功能恢复的影响

目的 观察骨髓间充质干细胞(MSCs)移植对脊髓损伤修复的促进作用.方法 用改良Allen法制作大鼠脊髓损伤动物模型.随机分成对照组(A组),脊髓损伤9 d后脊髓内微量注射生理盐水溶液5μl;骨髓间充质干细胞移植组(B组),脊髓损伤9 d后脊髓内注射骨髓间充质干细胞悬液5μl.移植后7、14、28 d采用斜板实验、脊髓运动功能BBB评分法观察大鼠运动功能恢复情况,脊髓诱发电位的检测观察神经功能恢复,苏木素-伊红(HE)染色观察脊髓损伤处空洞面积的改变情况,免疫组织化学法观察移植的骨髓间充质干细胞的存活及分化情况,损伤部位神经纤维的再生情况.结果 移植后28 d,两组斜板倾斜角度差异有统计学意义[A组(44.96±5.70)度,B组(53.19±6.51)度,P＜0.05];两组BBB评分差异有统计学意义[A组(6.8±1.2),B组(10.1±3.5),P＜0.05].同时,两组MEP潜伏期差异有统计学意义[A组(4.69±0.47)ms,B组(3.97±0.83)ms,P＜0.05],两组SEP潜伏期差异有统计学意义[A组(4.19±1.97)ms,B组(2.60±0.92)ms,P＜0.05].两组神经轴突计数差异有统计学意义[A组(32.8±6.1)条/mm2,B组(39.0±4.6)条/mm2,P＜0.05].实验组可见明显星形胶质细胞和神经纤维再生,脊髓损伤处的空洞面积明显减小.结论 骨髓间充质干细胞可在脊髓损伤处分化为神经元和神经胶质细胞,能够减小脊髓损伤处的空洞面积,促进受损轴突的再生和运动功能的恢复.     

SDF-1/CXCR4生物轴对胆囊癌细胞增殖及迁移的影响

目的 研究胆囊腺癌中趋化因子SDF-1及其受体CXCR4的表达情况,探讨其与胆囊腺癌临床病理特点及淋巴转移的关系.方法 采用免疫组化SP法检测41例胆囊腺癌中SDF-1及其受体CXCR4蛋白阳性表达情况,并分析其与临床病理参数的关系.结果 SDF-1在胆囊癌、胆囊炎、胆囊结石组和正常对照组胆囊黏膜中的表达率分别为68.3%(28/41)、6.7%(6/90)和5.0%(1/20),CXCR4的表达率分别为51.2%(21/41)、5.6%(5/90)和5.0%(1/20),SDF-1和CXCR4在胆囊癌与慢性胆囊炎、胆囊结石组胆囊黏膜、正常对照组胆囊黏膜中的阳性率比较,差异均有统计学意义(SDF-1:χ2=64.33,P＜0.001;CXCR4:χ2=42.52,P＜0.001),胆囊癌不同病理组织学分级、Nevin不同分期、组织学不同分化程度、有无淋巴结或远处转移组间的SDF-1和CXCR4的阳性率表达差异均有统计学意义(P均＜0.05),而不同性别、年龄、有无伴发胆囊结石组、肿瘤大小间SDF-1和CXCR4的阳性率表达差异均无统计学意义(P均＞0.05),胆囊癌组织中SDF-1阳性表达率(68.3%)与CX-CR4阳性表达率(51.2%)之间存在显著正相关(r=0.68,P＜0.01).结论 本研究表明,SDF-1/CXCR4生物轴与胆囊癌关系密切,提示可以通过干预SDF-1/CXCR4生物轴来治疗胆囊癌.     

红细胞生成素对慢性肾衰竭大鼠残肾组织归巢因子表达的影响

目的 观察红细胞生成素(EPO)对慢性肾衰竭(CRF)大鼠肾组织归巢因子表达的影响.方法 采用分阶段5/6肾切除制备大鼠CRF模型.实验动物随机分为3组:假手术组、CRF模型组和EPO治疗组.从第3周开始,治疗组大鼠每次皮下注射重组人EPO 50 IU/kg,每周3次,共6周.8周后检测各组大鼠血肌酐(Scr)、血尿素氮(BUN)、尿蛋白、血红蛋白(Hb)；采用实时荧光定量PCR、Western印迹和免疫组化方法检测残肾组织EPO及其受体(EPOR)、归巢因子及其受体(SDF-1、CXCR4、Ang-1、Tie2、SCF、c-Kit)的表达.结果 与模型组比较,EPO治疗可上调残肾组织归巢因子及其受体(SDF-1、CXCR4、Ang-1、Tie2、SCF、c-Kit)mRNA和蛋白的表达(均P＜0.05)；同时,EPO治疗还可上调残肾组织EPO及EPOR的mRNA和蛋白的表达(均P＜ 0.05).此外,EPO治疗还能下调大鼠Scr、BUN和尿蛋白水平(均P＜0.05),上调Hb水平(P＜0.05).结论 EPO能改善慢性肾衰竭大鼠的肾功能,这种作用可能与其激活残肾组织归巢因子而参与损伤肾脏的修复有关.     

基质细胞衍生因子联合外周血内皮祖细胞移植治疗裸鼠肢体缺血

目的 应用基质细胞衍生因子(SDF-1)结合外周血内皮祖细胞(EPCs)移植治疗裸鼠肢体缺血,探讨其促进血管新生的能力.方法 建立裸鼠后肢缺血模型,随机分为缺血对照组、EPCs移植组、SDF-1组、SDF-1联合EPCs移植组及SDF-1联合AMD3100处理后的EPCs移植组.移植第3、7天检测后肢腓肠肌CD34+VEGFR+细胞数,第28天检测缺血区新生血管密度.结果 移植第3天对照组、EPCs组、SDF-1组、SDF-1+EPCs组、SDF-1+AMD3100 EPCs组双阳性细胞数分别为0.00±0.00个/HP、5.30±0.65个/HP、0.00±0.00个/HP、10.31±0.63个/HP及1.86±0.17个/HP;第7天分别为0.00±0.00个/HP、7.05±0.18个/HP、0.00±0.00个/HP、11.81±0.53个/HP及2.83±0.48个/HP,第28天新生毛细血管数分别为3.00 ±0.13个/HP、6.15±0.04个/HP、6.20±0.10个/HP、10.65 ±0.08个/HP、6.21±0.08个/HP.SDF-1可增加缺血区CD34+ VEGFR+细胞浸润数量(P＜0.05),提高新生血管数量(P＜0.05),SDF-1联合外周血EPCs可进一步提高新生血管数量(P＜0.05).结论 SDF-1能够促进EPCs归巢,增强血管形成能力,促进血管新生;AMD3100能够阻断SDF-1的诱导迁移效应.

Abstract：

Objective To explore the effect of stromal cell-derived factor-1 ( SDF-1 ) in combination with transplantation of peripheral blood endothelial progenitor cells (EPCs) for the treatment of nude mice hindlimb ischemia. Methods Hindlimb ischemia model was established in nude mice, mice were then divided into five groups randomly: ischemic control group, peripheral blood EPCs transplantation group, SDF-1 local application group, SDF-1 combined with EPCs group, SDF-1 combined with AMD3100 treated EPCs group. Local CD34+VEGFR+ cells in the hind gastrocnemius were detected at day 3,7 after transplantation. The intensity of neovasculorization were evaluated at day 28. Results The double-positive cells number of control group, EPCs group, SDF-1 group, SDF-1 + EPCs group, SDF-1 + AMD3100 EPCs group were 0.00 ±0.00,5. 30 ±0.65,0.00 ±0.00,10. 31 ±0. 63,1. 86 ±0. 17 at day 3 and 0. 00 ±0. 00, 7.05 ±0. 18,0. 00 ±0. 00,11. 81 ±0. 53,2. 83 ±0. 48 at day 7. The number of new capillaries were 3. 00 ± 0.13,6.15 ± 0. 04,6. 20 ± 0. 10,10. 65 ± 0.08,6. 21 ±0. 08 at day 28. SDF-1 increased the CD34 + VEGFR+ cells (P ＜0. 05) and the number of new vessels (P ＜0.05). SDF-1 combined EPCs further increased the number of new vessels (P ＜ 0. 05 ). Conclusions SDF-1 enhances blood vessel formation and promotes angiogenesis by promoting EPCs homing, which could be blocked by AMD3100.     

骨髓间充质细胞和单个核细胞移植对糖尿病小鼠胰岛功能的影响

目的 比较骨髓间充质细胞移植和单个核细胞移植对糖尿病小鼠胰岛功能影响的差异.方法 建立糖尿病小鼠模型并分成3组:对照组(n=14)通过尾静脉注射磷酸盐缓冲液(PBS);单个核细胞组(n=14)通过尾静脉移植骨髓单个核细胞;间充质细胞组(n=14)通过尾静脉移植骨髓间充质细胞.观察移植后1周(n=6)和移植后6周(n=8),各组小鼠血糖的变化、胰岛数量、胰腺组织形态学特征及相关标记物的表达.结果 移植后1周,间充质细胞组小鼠血糖出现显著下降(16.6±1.6)mmol/L,与对照组(26.3±0.5)mmol/L和单个核细胞移植组(24.4±1.3)mmol/L比较差异有统计学意义(P＜0.05),并一直维持到移植后第6周,血糖下降到(16.5±1.5)mmol/L,与对照组(27.7±0.1)mmol/L比较差异有统计学意义(P＜0.05);移植后1周,间充质细胞组小鼠胰岛数目(21.2±1. 1)和胰岛β细胞数目(415.9±25.4)显著增加,与对照组(11.2±1.3)/(65.9±7.1)和单个核细胞组(12.2±1.3)/(64.1±6.5)比较差异均有统计学意义(P均＜0.05).单个核细胞组和间充质细胞组小鼠胰岛中均发现BrdU(+)Insulin(+)细胞和BrdU(+)Insulin(-)细胞.结论 骨髓间充质细胞移植改善糖尿病小鼠胰岛功能的效果优于骨髓单个核细胞移植.移植后胰岛的再生既来源于胰岛β细胞的增殖,也可能来源于胰岛干细胞的分化.

Abstract：

Objective To compare the different effects of bone marrow mononuclear cells vs mesenchymal cells transplantation on islets function of diabetic mice. Methods Mouse diabetic models were created by multiply peritoneal injection of low-dose streptozotocin (STZ) and divided into three groups:control group ( n = 14) , bone marrow mononuclear cells group ( n = 14) , and bone marrow mesenchymal cells group (n = 14). Blood glucose was measured weekly after transplantation by glucometer. Histochem istry and immunofluorescence were performed to characterize pancreatic histology, morphology and markers expressed in receipt pancreas. Results Compared with control group and bone marrow mesenchymal cells group, blood glucose levels in bone marrow mesenchymal cells group were significantly reduced at first week after transplantation[( 16. 6 ± 1.6 ) vs ( 26. 3 ± 0. 5 ) / ( 24. 4 ± 1.3 ) mmol/L, P ＜ 0. 05]and sustained to reduce at 6th week after transplantation[( 16. 5 ± 1.5 ) vs ( 27.7 ± 0. 1 ) mmol/L in control group,P＜0. 05]. One week after transplantation, the islets number in bone marrow mesenchymal cells group was larger than in control group ( 21.2 ± 1. 1vs 11.2 ± 1.3, P ＜ 0. 05 ) and bone marrow mononuclear cells group ( 21.2 ± 1. 1vs 12. 2 ± 1.3 ,P ＜0. 05 ). One weeks after transplantation, the beta cell number in bone marrow mesenchymal cells group was larger than in control group (415.9 ± 25.4 vs 65.9 ±7. 1,P＜0.05) and bone marrow mononuclear cells group (415.9 ±25.4 vs 64. 1 ±6.5,P＜0.05). In bone marrow mononuclear cells and bone marrow mesenchymal cells groups, there were several BrdU ( + )Insulin( - ) cells and BrdU( + )Insulin( - ) cells in the islets. Conclusion The effect of bone marrow mesenchymal cells transplantation to improve diabetic islet function is more satisfactory than bone marrow mononuclear cells transplantation. Bone marrow mesenchymal cells transplantation can initiate pancreatic islets β cells regeneration by both proliferation of β cells and differentiation of pancreatic stem cells.     

与大鼠胰岛细胞联合移植的骨髓间充质干细胞的免疫调节作用

目的 研究同种异体或自体的大鼠骨髓间充质干细胞(MSC)与胰岛肝内联合移植对胰岛移植物的免疫调节作用及其机制.方法 以链佐星制备Lewis大鼠的糖尿病模型,作为胰岛移植受者,分为3组:单纯移植组大鼠经门静脉单独移植SD大鼠胰岛6000 IEQ/kg;同系MSC组大鼠经门静脉共同移植1×109/L的Lewis大鼠MSC 1 ml与SD大鼠胰岛6000 IEQ/kg;同种MSC组大鼠经门静脉共同移植1×109/L的SD大鼠MSC 1 ml与SD大鼠胰岛6000 IEQ/kg.检测受鼠的血糖变化,术后1、3 d大鼠外周血γ干扰素(IFN-γ)、白细胞介素(IL)-2、IL-4和IL-10的含量.结果 3组大鼠术后第1天血糖均下降到13.9 mmol/L以下.同系MSC组移植物存活时间为(11.38±4.03)d,同种MSC组为(5.50±2.07)d,单纯移植组为(2.88±1.25)d(P＜0.01).术后1、3 d,单纯移植组IFN-γ和IL-2的含量显著高于同系MSC组和同种MSC组(P＜0.01),同种MSC组IFN-γ和IL-2的含量高于同系MSC组(P＜0.05);单纯移植组IL-10的含量低于同系MSC组和同种MSC组(P＜0.01),同系MSC组IL-10的含量与司种MSC组相比较,差异无统计学意义(P＞0.05);各组IL-4含量的差异无统计学意义(P＞0.05).结论 MSC与同种胰岛共移植可以延长胰岛移植物存活时间,应用同系MSC的效果优于同种异体MSC.共移植的MSC主要通过减少TH1类细胞因子(IFN-y和IL-2)的表达使受者TH1/TH2平衡向TH2方向偏移.

Abstract：

Objective To compare the immune regulation of syngenic and allogenic mesenchymal stem cells (MSCs) in the transplantation combined with islets. Methods After induction of diabetes in 30 Lewis rats with streptozotocin (STZ), the recipient Lewis rats received islets from SD rats combined with syngenic (group B) or allogenic (group C) MSCs injection via the portal vein. The group of islets transplanted alone served as control (group A). The survival time of grafts in all groups was assessed by the level of blood glucose. ELISA was used to detect the levels of interferon-γ (IFN-γ), interleukin 2 (IL-2), IL-4 and IL-10 in the peripheral blood on the 1st and 3rd day after transplantation. Results The blood glucose levels in all three groups were decreased in a normal range (13. 9 mmol/L) and the survival time of grafts in group B (11.38 ± 4. 03 days) was significantly longer than in group C (5. 50± 2. 07 days) as well as group A (2. 88 ± 1.25 days). On the 1 st and 3rd day after transplantation, the levels of TH 1 cytokines IFN-γ and IL-2 in group A were significantly higher than in groups C and B (P＜0.05). Meanwhile the levels of TH 2 cytokine IL-10were increased in group B, but there was no significant difference between groups A and C (P＞0.05). The levels of IL-4 had no significant difference among these three groups (P ＞ 0.05).Conclusion Islet transplantation combined with MSCs could prolong the survival time of grafts.Syngenic MSCs, superior to allogenic ones, were more effective in changing the balance of TH1/TH2to TH2. Decreased expression of TH1 cytokine (IFN-γ, IL-2), which was closely related to the induction of immune tolerance, was beneficial to the long-term survival of grafts.     

大鼠骨髓间充质细胞经SDF-1预处理后移植对急性心肌梗死的治疗效果

目的 探讨大鼠骨髓间充质细胞(BMSC)经基质细胞衍生因子-1(SDF-1)预处理后移植对急性心肌梗死(AMI)的治疗效果.方法 (1)全骨髓贴壁法培养大鼠BMSC;逆转录聚合酶链反应(RT-PCR)和免疫组织化学法检测BMSC趋化因子受体(CXCR4)基因的表达;将BMSC分别与10和100μg/L的SDF-1作用24 h后,在无氧、无血清条件下培养6 h,流式细胞仪和末端标记(TUNEL)法检测细胞凋亡率.(2)建立大鼠急性心肌梗死模型,将经100μg/L SDF-1预处理的BMSC和未经处理的BMSC植入大鼠梗死心肌周边,2周后采用超声心动图观察心脏功能的变化.结果 BMSC表达CXCR4基因;在无氧和无血清条件下,经SDF-1预处理的BMSC凋亡率与对照组比较明显降低(P<0.05),而经100 μg/L SDF-1预处理的BMSC凋亡率最低.成功建立大鼠急性心肌梗死模型;与未经处理的BMSC移植比较,经SDF-1预处理的BMSC移植后改善心肌梗死大鼠的心功能作用更为明显(P<0.05).结论 用SDF-1预处理大鼠BMSC能抑制其在无氧和无血清条件下的凋亡.用SDF-1预处理BMSC能增强其移植后治疗大鼠急性心肌梗死的效果.     

SDF-1/CXCR4信号轴调控骨髓间充质干细胞向哮喘小鼠肺组织的迁移

目的：探讨基质细胞衍生因子-1（SDF-1）/CXCR4轴在外源性骨髓间充质干细胞（MSC）向哮喘模型小鼠肺组织迁移的作用.方法：无菌条件下取GFP转基因小鼠骨髓MSC,体外扩增,鉴定.采用Transwell培养系统,观察0、50、100、150和200 ng/ml SDF-1和CXCR4阻断剂AMD3100对MSC体外定向迁移的影响.取60只雌性BALB/c小鼠,随机分为6组（n=10）：PBS非哮喘组、MSC非哮喘组、PBS哮喘组、MSC哮喘组、SDF-1处理哮喘组、AMD3100处理哮喘组.哮喘组采用哮喘致敏液0.2 ml（含100 μg卵白蛋白）致敏,并使用卵白蛋白雾化吸入激发哮喘.非哮喘组在致敏和激发时均予以PBS处理.MSC处理组于哮喘激发前移植外源性MSC.SDF-1处理哮喘组在MSC移植前气管内注入SDF-1,AMD3100处理哮喘组注入AMD3100预先孵育的MSC.PBS哮喘组注射等量的PBS液.采用Westernablot和RT-PCR方法检测肺组织中SDF-1的表达水平,通过荧光显微镜观察表达GFP的外源性MSC在哮喘小鼠肺组织中的分布情况,比较SDF-1和CXCR4阻断剂AMD3100干预对MSC向肺组织迁移的影响.结果：Transwell实验显示MSC的迁移水平与SDF-1（0~150军ng/ml）成浓度依赖性.与正常小鼠比较,哮喘小鼠肺组织SDF-1表达增强.与MSC非哮喘组比较,MSC哮喘组小鼠肺部有更多MSC聚集.在哮喘肺组织中增加外源性SDF-1能够促进MSC向肺组织迁移.通过AMD3100阻断MSC的CXCR4可以明显减少MSC向肺组织的迁移水平.结论：SDF-1/CXCR4轴参与了MSC迁移到哮喘小鼠肺组织的过程.     

异体大鼠骨髓单个核细胞胰岛细胞移植治疗糖尿病

目的 观察异体骨髓单个核细胞和胰岛细胞通过肝脏和静脉途径移植后对糖尿病大鼠的治疗作用.方法 密度梯度离心法分离胰岛细胞,淋巴细胞分离液分离骨髓单个核细胞,28只糖尿病大鼠模型随机分为A、B、C、D组,A组在肝脏被膜下多点注射1000个胰岛细胞,B组在体外将1000个胰岛细胞和1×107个骨髓单个核细胞混合后在肝脏被膜下多点注射,C组通过尾静脉注射1000个胰岛细胞,D组在体外将1000个胰岛细胞和1×107个骨髓单个核细胞混合后通过尾静脉注射,移植后于不同时间点尾静脉测定随机血糖,比较不同细胞组合和移植途径之间对糖尿病的治疗作用.结果 A、B组血糖于术后3 d内开始下降,A组血糖可降至正常水平(7.98±2.28)mmol/L,血糖维持正常水平(3.71±0.95)d,B组降至(7.72±1. 75)mmol/L可维持(4.86±1.06)d,静脉移植组血糖于术后4 d内降至正常(7.35±1.40)mmol/L,可维持(7.85±1.46)d,D组静脉注射胰岛于4 d起效(7.00±0.83)mmol/L,血糖可降至正常水平可维持(14.10±1.21)d,各组间血糖随时间变化的趋势及维持正常水平的时间具有统计学意义(P＜0.05).结论 骨髓单个核细胞和胰岛混合细胞通过尾静脉移植对大鼠血糖维持正常时间最长,血糖控制水平最理想.