移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

Objective To investigate the expression and significance of local Th1 and Th2 cytokines in the grafts of pig simultaneous pancreas-kidney with portal versus systemic venous drainage. Methods Fifty-eight non-related and first hybrid landraces were divided into Sham (n = 10), PE (portal-enteric drainage) (n =24) and SE ( systemic-enteric drainage) (n =24) groups. Type 1 diabetic mellitus was induced by whole pancreatectomy ,and right-side nephrectomy was also performed in the recipients. The pigs in sham group were only subjected to ]aparotomy. The portal vein was anastomosed to superior mesenteric vein of the recipient in PE group or to the inferior hepatic cava of the recipients in SE group. The tissues of the graft pancreas and kidney were obtained by surgery for detecting the expression of mRNA and protein of IFN-γand IL4 by RT-PCR and ELISA on the day 3,and 7 after transplantation. Results Compared with sham group,the local expression of mRNA and protein of IFN in the graft pancreas and kidney of both SE and PE groups was significantly up-regulated ( P < 0. 05 ) ( On the day 7, protein expression in sham, SE, and PE groups :in the graft pancreas, [ (3.36±0.25 ), (7.72±0.67 ), and ( 6.52±0.34 ) pg/mg protein ];in the graft kidney, [ ( 3.66±0.28 ), ( 10.83±1.48 ), and (7.79±0.60) pg/mg protein ]. The local expression of mRNA and protein of IL-4 was significantly up-regulated in PE group (P < 0.05 ), but down-regulated or no expression was detected in group SE (on the day 7, protein expression in sham, SE, and PE groups:in the graft pancreas, [ (7.18±0.16), (6.10±0.16) ,and (20.66±1.47) pg/mg protein];in the graft kidney, [ (5.74±0.48) ,(10.38±0.92) ,and ( 19.66±1.57) pg/mg protein]. Conclusion That the blood flow of graft pancreas passes by liver in SPK with PVD can induce a Th1 to Th2 inmaune shift. This may be one of mechanisms of immunologic benefits of PVD.     

移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

Objective To investigate the expression and significance of local Th1 and Th2 cytokines in the grafts of pig simultaneous pancreas-kidney with portal versus systemic venous drainage. Methods Fifty-eight non-related and first hybrid landraces were divided into Sham (n = 10), PE (portal-enteric drainage) (n =24) and SE ( systemic-enteric drainage) (n =24) groups. Type 1 diabetic mellitus was induced by whole pancreatectomy ,and right-side nephrectomy was also performed in the recipients. The pigs in sham group were only subjected to ]aparotomy. The portal vein was anastomosed to superior mesenteric vein of the recipient in PE group or to the inferior hepatic cava of the recipients in SE group. The tissues of the graft pancreas and kidney were obtained by surgery for detecting the expression of mRNA and protein of IFN-γand IL4 by RT-PCR and ELISA on the day 3,and 7 after transplantation. Results Compared with sham group,the local expression of mRNA and protein of IFN in the graft pancreas and kidney of both SE and PE groups was significantly up-regulated ( P < 0. 05 ) ( On the day 7, protein expression in sham, SE, and PE groups :in the graft pancreas, [ (3.36±0.25 ), (7.72±0.67 ), and ( 6.52±0.34 ) pg/mg protein ];in the graft kidney, [ ( 3.66±0.28 ), ( 10.83±1.48 ), and (7.79±0.60) pg/mg protein ]. The local expression of mRNA and protein of IL-4 was significantly up-regulated in PE group (P < 0.05 ), but down-regulated or no expression was detected in group SE (on the day 7, protein expression in sham, SE, and PE groups:in the graft pancreas, [ (7.18±0.16), (6.10±0.16) ,and (20.66±1.47) pg/mg protein];in the graft kidney, [ (5.74±0.48) ,(10.38±0.92) ,and ( 19.66±1.57) pg/mg protein]. Conclusion That the blood flow of graft pancreas passes by liver in SPK with PVD can induce a Th1 to Th2 inmaune shift. This may be one of mechanisms of immunologic benefits of PVD.     

移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

Objective To investigate the expression and significance of local Th1 and Th2 cytokines in the grafts of pig simultaneous pancreas-kidney with portal versus systemic venous drainage. Methods Fifty-eight non-related and first hybrid landraces were divided into Sham (n = 10), PE (portal-enteric drainage) (n =24) and SE ( systemic-enteric drainage) (n =24) groups. Type 1 diabetic mellitus was induced by whole pancreatectomy ,and right-side nephrectomy was also performed in the recipients. The pigs in sham group were only subjected to ]aparotomy. The portal vein was anastomosed to superior mesenteric vein of the recipient in PE group or to the inferior hepatic cava of the recipients in SE group. The tissues of the graft pancreas and kidney were obtained by surgery for detecting the expression of mRNA and protein of IFN-γand IL4 by RT-PCR and ELISA on the day 3,and 7 after transplantation. Results Compared with sham group,the local expression of mRNA and protein of IFN in the graft pancreas and kidney of both SE and PE groups was significantly up-regulated ( P < 0. 05 ) ( On the day 7, protein expression in sham, SE, and PE groups :in the graft pancreas, [ (3.36±0.25 ), (7.72±0.67 ), and ( 6.52±0.34 ) pg/mg protein ];in the graft kidney, [ ( 3.66±0.28 ), ( 10.83±1.48 ), and (7.79±0.60) pg/mg protein ]. The local expression of mRNA and protein of IL-4 was significantly up-regulated in PE group (P < 0.05 ), but down-regulated or no expression was detected in group SE (on the day 7, protein expression in sham, SE, and PE groups:in the graft pancreas, [ (7.18±0.16), (6.10±0.16) ,and (20.66±1.47) pg/mg protein];in the graft kidney, [ (5.74±0.48) ,(10.38±0.92) ,and ( 19.66±1.57) pg/mg protein]. Conclusion That the blood flow of graft pancreas passes by liver in SPK with PVD can induce a Th1 to Th2 inmaune shift. This may be one of mechanisms of immunologic benefits of PVD.     

移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

Objective To investigate the expression and significance of local Th1 and Th2 cytokines in the grafts of pig simultaneous pancreas-kidney with portal versus systemic venous drainage. Methods Fifty-eight non-related and first hybrid landraces were divided into Sham (n = 10), PE (portal-enteric drainage) (n =24) and SE ( systemic-enteric drainage) (n =24) groups. Type 1 diabetic mellitus was induced by whole pancreatectomy ,and right-side nephrectomy was also performed in the recipients. The pigs in sham group were only subjected to ]aparotomy. The portal vein was anastomosed to superior mesenteric vein of the recipient in PE group or to the inferior hepatic cava of the recipients in SE group. The tissues of the graft pancreas and kidney were obtained by surgery for detecting the expression of mRNA and protein of IFN-γand IL4 by RT-PCR and ELISA on the day 3,and 7 after transplantation. Results Compared with sham group,the local expression of mRNA and protein of IFN in the graft pancreas and kidney of both SE and PE groups was significantly up-regulated ( P < 0. 05 ) ( On the day 7, protein expression in sham, SE, and PE groups :in the graft pancreas, [ (3.36±0.25 ), (7.72±0.67 ), and ( 6.52±0.34 ) pg/mg protein ];in the graft kidney, [ ( 3.66±0.28 ), ( 10.83±1.48 ), and (7.79±0.60) pg/mg protein ]. The local expression of mRNA and protein of IL-4 was significantly up-regulated in PE group (P < 0.05 ), but down-regulated or no expression was detected in group SE (on the day 7, protein expression in sham, SE, and PE groups:in the graft pancreas, [ (7.18±0.16), (6.10±0.16) ,and (20.66±1.47) pg/mg protein];in the graft kidney, [ (5.74±0.48) ,(10.38±0.92) ,and ( 19.66±1.57) pg/mg protein]. Conclusion That the blood flow of graft pancreas passes by liver in SPK with PVD can induce a Th1 to Th2 inmaune shift. This may be one of mechanisms of immunologic benefits of PVD.     

移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

Objective To investigate the expression and significance of local Th1 and Th2 cytokines in the grafts of pig simultaneous pancreas-kidney with portal versus systemic venous drainage. Methods Fifty-eight non-related and first hybrid landraces were divided into Sham (n = 10), PE (portal-enteric drainage) (n =24) and SE ( systemic-enteric drainage) (n =24) groups. Type 1 diabetic mellitus was induced by whole pancreatectomy ,and right-side nephrectomy was also performed in the recipients. The pigs in sham group were only subjected to ]aparotomy. The portal vein was anastomosed to superior mesenteric vein of the recipient in PE group or to the inferior hepatic cava of the recipients in SE group. The tissues of the graft pancreas and kidney were obtained by surgery for detecting the expression of mRNA and protein of IFN-γand IL4 by RT-PCR and ELISA on the day 3,and 7 after transplantation. Results Compared with sham group,the local expression of mRNA and protein of IFN in the graft pancreas and kidney of both SE and PE groups was significantly up-regulated ( P < 0. 05 ) ( On the day 7, protein expression in sham, SE, and PE groups :in the graft pancreas, [ (3.36±0.25 ), (7.72±0.67 ), and ( 6.52±0.34 ) pg/mg protein ];in the graft kidney, [ ( 3.66±0.28 ), ( 10.83±1.48 ), and (7.79±0.60) pg/mg protein ]. The local expression of mRNA and protein of IL-4 was significantly up-regulated in PE group (P < 0.05 ), but down-regulated or no expression was detected in group SE (on the day 7, protein expression in sham, SE, and PE groups:in the graft pancreas, [ (7.18±0.16), (6.10±0.16) ,and (20.66±1.47) pg/mg protein];in the graft kidney, [ (5.74±0.48) ,(10.38±0.92) ,and ( 19.66±1.57) pg/mg protein]. Conclusion That the blood flow of graft pancreas passes by liver in SPK with PVD can induce a Th1 to Th2 inmaune shift. This may be one of mechanisms of immunologic benefits of PVD.     

移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

Objective To investigate the expression and significance of local Th1 and Th2 cytokines in the grafts of pig simultaneous pancreas-kidney with portal versus systemic venous drainage. Methods Fifty-eight non-related and first hybrid landraces were divided into Sham (n = 10), PE (portal-enteric drainage) (n =24) and SE ( systemic-enteric drainage) (n =24) groups. Type 1 diabetic mellitus was induced by whole pancreatectomy ,and right-side nephrectomy was also performed in the recipients. The pigs in sham group were only subjected to ]aparotomy. The portal vein was anastomosed to superior mesenteric vein of the recipient in PE group or to the inferior hepatic cava of the recipients in SE group. The tissues of the graft pancreas and kidney were obtained by surgery for detecting the expression of mRNA and protein of IFN-γand IL4 by RT-PCR and ELISA on the day 3,and 7 after transplantation. Results Compared with sham group,the local expression of mRNA and protein of IFN in the graft pancreas and kidney of both SE and PE groups was significantly up-regulated ( P < 0. 05 ) ( On the day 7, protein expression in sham, SE, and PE groups :in the graft pancreas, [ (3.36±0.25 ), (7.72±0.67 ), and ( 6.52±0.34 ) pg/mg protein ];in the graft kidney, [ ( 3.66±0.28 ), ( 10.83±1.48 ), and (7.79±0.60) pg/mg protein ]. The local expression of mRNA and protein of IL-4 was significantly up-regulated in PE group (P < 0.05 ), but down-regulated or no expression was detected in group SE (on the day 7, protein expression in sham, SE, and PE groups:in the graft pancreas, [ (7.18±0.16), (6.10±0.16) ,and (20.66±1.47) pg/mg protein];in the graft kidney, [ (5.74±0.48) ,(10.38±0.92) ,and ( 19.66±1.57) pg/mg protein]. Conclusion That the blood flow of graft pancreas passes by liver in SPK with PVD can induce a Th1 to Th2 inmaune shift. This may be one of mechanisms of immunologic benefits of PVD.     

移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

Objective To investigate the expression and significance of local Th1 and Th2 cytokines in the grafts of pig simultaneous pancreas-kidney with portal versus systemic venous drainage. Methods Fifty-eight non-related and first hybrid landraces were divided into Sham (n = 10), PE (portal-enteric drainage) (n =24) and SE ( systemic-enteric drainage) (n =24) groups. Type 1 diabetic mellitus was induced by whole pancreatectomy ,and right-side nephrectomy was also performed in the recipients. The pigs in sham group were only subjected to ]aparotomy. The portal vein was anastomosed to superior mesenteric vein of the recipient in PE group or to the inferior hepatic cava of the recipients in SE group. The tissues of the graft pancreas and kidney were obtained by surgery for detecting the expression of mRNA and protein of IFN-γand IL4 by RT-PCR and ELISA on the day 3,and 7 after transplantation. Results Compared with sham group,the local expression of mRNA and protein of IFN in the graft pancreas and kidney of both SE and PE groups was significantly up-regulated ( P < 0. 05 ) ( On the day 7, protein expression in sham, SE, and PE groups :in the graft pancreas, [ (3.36±0.25 ), (7.72±0.67 ), and ( 6.52±0.34 ) pg/mg protein ];in the graft kidney, [ ( 3.66±0.28 ), ( 10.83±1.48 ), and (7.79±0.60) pg/mg protein ]. The local expression of mRNA and protein of IL-4 was significantly up-regulated in PE group (P < 0.05 ), but down-regulated or no expression was detected in group SE (on the day 7, protein expression in sham, SE, and PE groups:in the graft pancreas, [ (7.18±0.16), (6.10±0.16) ,and (20.66±1.47) pg/mg protein];in the graft kidney, [ (5.74±0.48) ,(10.38±0.92) ,and ( 19.66±1.57) pg/mg protein]. Conclusion That the blood flow of graft pancreas passes by liver in SPK with PVD can induce a Th1 to Th2 inmaune shift. This may be one of mechanisms of immunologic benefits of PVD.     

移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

Objective To investigate the expression and significance of local Th1 and Th2 cytokines in the grafts of pig simultaneous pancreas-kidney with portal versus systemic venous drainage. Methods Fifty-eight non-related and first hybrid landraces were divided into Sham (n = 10), PE (portal-enteric drainage) (n =24) and SE ( systemic-enteric drainage) (n =24) groups. Type 1 diabetic mellitus was induced by whole pancreatectomy ,and right-side nephrectomy was also performed in the recipients. The pigs in sham group were only subjected to ]aparotomy. The portal vein was anastomosed to superior mesenteric vein of the recipient in PE group or to the inferior hepatic cava of the recipients in SE group. The tissues of the graft pancreas and kidney were obtained by surgery for detecting the expression of mRNA and protein of IFN-γand IL4 by RT-PCR and ELISA on the day 3,and 7 after transplantation. Results Compared with sham group,the local expression of mRNA and protein of IFN in the graft pancreas and kidney of both SE and PE groups was significantly up-regulated ( P < 0. 05 ) ( On the day 7, protein expression in sham, SE, and PE groups :in the graft pancreas, [ (3.36±0.25 ), (7.72±0.67 ), and ( 6.52±0.34 ) pg/mg protein ];in the graft kidney, [ ( 3.66±0.28 ), ( 10.83±1.48 ), and (7.79±0.60) pg/mg protein ]. The local expression of mRNA and protein of IL-4 was significantly up-regulated in PE group (P < 0.05 ), but down-regulated or no expression was detected in group SE (on the day 7, protein expression in sham, SE, and PE groups:in the graft pancreas, [ (7.18±0.16), (6.10±0.16) ,and (20.66±1.47) pg/mg protein];in the graft kidney, [ (5.74±0.48) ,(10.38±0.92) ,and ( 19.66±1.57) pg/mg protein]. Conclusion That the blood flow of graft pancreas passes by liver in SPK with PVD can induce a Th1 to Th2 inmaune shift. This may be one of mechanisms of immunologic benefits of PVD.     

移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

Objective To investigate the expression and significance of local Th1 and Th2 cytokines in the grafts of pig simultaneous pancreas-kidney with portal versus systemic venous drainage. Methods Fifty-eight non-related and first hybrid landraces were divided into Sham (n = 10), PE (portal-enteric drainage) (n =24) and SE ( systemic-enteric drainage) (n =24) groups. Type 1 diabetic mellitus was induced by whole pancreatectomy ,and right-side nephrectomy was also performed in the recipients. The pigs in sham group were only subjected to ]aparotomy. The portal vein was anastomosed to superior mesenteric vein of the recipient in PE group or to the inferior hepatic cava of the recipients in SE group. The tissues of the graft pancreas and kidney were obtained by surgery for detecting the expression of mRNA and protein of IFN-γand IL4 by RT-PCR and ELISA on the day 3,and 7 after transplantation. Results Compared with sham group,the local expression of mRNA and protein of IFN in the graft pancreas and kidney of both SE and PE groups was significantly up-regulated ( P < 0. 05 ) ( On the day 7, protein expression in sham, SE, and PE groups :in the graft pancreas, [ (3.36±0.25 ), (7.72±0.67 ), and ( 6.52±0.34 ) pg/mg protein ];in the graft kidney, [ ( 3.66±0.28 ), ( 10.83±1.48 ), and (7.79±0.60) pg/mg protein ]. The local expression of mRNA and protein of IL-4 was significantly up-regulated in PE group (P < 0.05 ), but down-regulated or no expression was detected in group SE (on the day 7, protein expression in sham, SE, and PE groups:in the graft pancreas, [ (7.18±0.16), (6.10±0.16) ,and (20.66±1.47) pg/mg protein];in the graft kidney, [ (5.74±0.48) ,(10.38±0.92) ,and ( 19.66±1.57) pg/mg protein]. Conclusion That the blood flow of graft pancreas passes by liver in SPK with PVD can induce a Th1 to Th2 inmaune shift. This may be one of mechanisms of immunologic benefits of PVD.     

人血管内皮生长因子165基因转染人骨髓间充质干细胞的实验研究

Objective To establish an effective method of transfecting human marrow mesenchymal stem cells (MSC) with human vascular endothelial growth factor 165 ( VEGF 165) gene. Methods MSCs isolated and cultured in vitro were divided into transfection group (pShuttle-CMV/VEGF 165 plasmid was transfected into MSCs through liposome-mediating method), empty plasmid group (pShuttle-CMV vehi-cle was transfected into MSCs as control), liposome group (liposome was transfected into MSCs as control) and control group(normal culture). Expressions of Mrna and protein of MSCs were determined by RT-PCR, enzyme-linked immunosorbent assay and Western Blot. Sensitivity to MSCs on VEGF plasmid transfec-tion was detected by MTT test. Results Expression level of VEGF 165 gene Mrna in transfection group, empty plasmid group, liposome group, and control group was respectively 0.89 ± 0.03, 0. 34 ± 0.04, 0.40 ± 0.03, and 0.30 ± 0.03, and the difference between transfection group and the other three groups was statistically significant ( P <0. 01 ). Content of VEGF protein in transfection group, empty plasmid group, liposome group, and control group was respectively (778 ± 35 ), (543 ± 24), (561 ± 28), (571 ± 23) pg/Ml, and the difference between transfection group and the other three groups was statistically significant ( P <0.01 ). In the transfection group, expression level of VEGF protein peaked on 7th day after transfec-tion, which was decreased gradually later. In transfection group, expression level of V EGF 165 protein was obviously higher than that of the other three groups ( P <0. 01 ), and no inhibitory effect of VEGF plasmid transfection on MSCs proliferation was found. Conclusions The method for transfecting human VEGF 165 gene into MSCs is established in this research, through which target gene and protein can express effectively.     

移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

目的 探讨Th1和Th2型细胞因子在猪胰肾联合移植门脉(PVD)和体静脉(SVD)回流术式移植物局部表达及意义.方法 58例四川当地杂交第1代长白猪,分成假手术组(10例)、PE组(24例)和SE组(24例).假手术组行开腹术,PE组和SE组切除受者胰腺制成Ⅰ型糖尿病模型,并切除右肾.PE组门静脉与受者肠系膜上静脉吻合,SE组门静脉与受者肝下下腔静脉吻合,外分泌均采用肠道引流.术后第3、7天开腹取移植胰腺、肾脏组织,逆转录-聚合酶链反应(RT-PCR)和ELISA检测移植物局部Th1型细胞因子IFN-γ和Th2型细胞因子白细胞介素(IL)-4mRNA及蛋白的表达.结果 术后第3、7天,与对照组比较,移植肾和胰腺IFN-γ mRNA及蛋白在SE组和PE组均呈高表达(P<0.05),SE组高于PE组(P<0.05)(第7天,蛋白在对照组比SE组比PE组表达:移植胰腺,[(3.36±0.25)比(7.72±0.67)比(6.52±0.34)pg/mg蛋白],移植肾[(3.66±0.28)比(10.83±1.48)比(7.79±0.60)pg/mg蛋白].移植胰腺和肾IL-4 mRNA及蛋白在SE组和假手术组呈低表达,PE组呈高表达(P<0.05)(第7天,蛋白在对照组比SE组比PE组表达:移植胰腺,[(7.18±0.16)比(6.10±0.16)比(20.66±1.47)pg/mg蛋白];移植肾,[(5.74±0.48)比(10.38±0.92)比(19.66±1.57)pg/mg蛋白].结论 胰肾联合移植PVD术式移植物血流经过肝脏后可诱导,Th1细胞向Th2细胞偏离,是PVD免疫学优越性的可能机制.