神经生长因子基因修饰的骨髓间充质干细胞在糖尿病大鼠膀胱中的生长及表达

目的 将转入神经生长因子(NGF)的大鼠骨髓间充质干细胞(MSC)移植于糖尿病大鼠膀胱平滑肌组织内,观察MSC在膀胱组织内的存活和NGF基因的表达情况.方法 糖尿病组(30只)大鼠按链脲佐菌素(STZ)60 mg/kg行单次腹腔注射,对照组(15只)腹腔注射等体积的柠檬酸缓冲液.实验分组:对照组(正常大鼠膀胱)、发病组(糖尿病大鼠膀胱)、治疗组(糖尿病大鼠膀胱内移植转染NGF基因的MSC).溴苷法示踪NGF基因修饰的MSC在大鼠膀胱内的存活情况;RTPCR、ELISA法检测NGF基因在糖尿病大鼠膀胱内的表达情况.结果 单次腹腔注射STZ造模成功,8周后血糖仍处高位.NGF基因修饰的大鼠MSC移植入糖尿病大鼠膀胱内4周仍存活.ELISA检测结果显示对照组、发病组、治疗组大鼠膀胱NGF蛋白含量分别为(114±3)、(70±2)、(110±2)pg/ml,RT-PCR检测mRNA表达量分别为0.183±0.004、0.032±0.139、0.130±0.165.发病组与对照组相比NGF表达下降(P＜0.05),治疗组与发病组相比NGF表达上升(P＜0.05).结论转入NGF基因的大鼠MSC能在糖尿病大鼠膀胱中存活并稳定表达.

Abstract：

Objective To confirm that rat bone marrow mesenchymal stem cells (MSC) transfected with nerve growth factor (NGF) gene in the bladder tissue of diabetic rats bladder tissues can survive and stably express NGF. Methods A diabetic rat model was constructed. The BrdU-labelled MSC transfected with NGF gene were transplanted into the diabetic rats bladder tissues. BrdUlabelled immunohistochemistry was used to observe the growth of MSC transfected with NGF gene in the diabetic rats bladder tissues. The expression of NGF mRNA and protein were checked by RT-PCR and ELISA. Results A diabetic rat model was successfully built by a single intraperitoneal injectionof STZ. The blood glucose was still high after 8 weeks. NGF gene modified MSC could be detected in the bladder of diabetic rats by BrdU-labelled immunohistochemistry. The concentration of NGF in the control group, disease group and treatment group were ( 114 ± 3), ( 70 ± 2), ( 110 ± 2) pg/ml by ELISA and mRNA quantity by RT-PCR were 0. 183±0. 004, 0. 032±0. 139, 0. 130±0. 165, respectively. Compared with the control group, the expression of NGF gene was decreased (P＜0. 05) in the incidence group. The expression of NGF gene was increased (P＜0. 05) in the treatment group compared with the disease group. Conclusions The NGF gene-modified MSC could survive in diabetic rats bladder tissues. The NGF gene in MSC could stably express in diabetic rats bladder tissues.

Growth and expression of rat bone marrow mesenchymal stem cells modified with nerve growth factor in the bladder

Objective To confirm that rat bone marrow mesenchymal stem cells (MSC) transfected with nerve growth factor (NGF) gene in the bladder tissue of diabetic rats bladder tissues can survive and stably express NGF. Methods A diabetic rat model was constructed. The BrdU-labelled MSC transfected with NGF gene were transplanted into the diabetic rats bladder tissues. BrdUlabelled immunohistochemistry was used to observe the growth of MSC transfected with NGF gene in the diabetic rats bladder tissues. The expression of NGF mRNA and protein were checked by RT-PCR and ELISA. Results A diabetic rat model was successfully built by a single intraperitoneal injectionof STZ. The blood glucose was still high after 8 weeks. NGF gene modified MSC could be detected in the bladder of diabetic rats by BrdU-labelled immunohistochemistry. The concentration of NGF in the control group, disease group and treatment group were ( 114 ± 3), ( 70 ± 2), ( 110 ± 2) pg/ml by ELISA and mRNA quantity by RT-PCR were 0. 183±0. 004, 0. 032±0. 139, 0. 130±0. 165, respectively. Compared with the control group, the expression of NGF gene was decreased (P＜0. 05) in the incidence group. The expression of NGF gene was increased (P＜0. 05) in the treatment group compared with the disease group. Conclusions The NGF gene-modified MSC could survive in diabetic rats bladder tissues. The NGF gene in MSC could stably express in diabetic rats bladder tissues.