骨髓间充质干细胞移植对博莱霉素致小鼠肺纤维化形成期干预的研究
发布时间：2018-07-13 21:57
【摘要】：目的：研究移植外源性骨髓间充质干细胞(BMSCs)对博莱霉素致小鼠晚期肺纤维化即肺纤维化形成期的干预作用。 方法：全骨髓贴壁细胞分离培养法分离、培养雄性6-8周龄ICR小鼠BMSCs。本实验中设计4个分组，共160只实验小鼠，将这些小鼠随机分为：阴性对照组（NC组），阳性对照组（PC组），BMSCs干预组A（A组），BMSCs干预组B（B组）。NC组于经鼻腔滴入50ulPBS作为阴性对照，PC组、A组、B组则按6ug/g的剂量经鼻腔滴入50ul博莱霉素制成小鼠的肺纤维化模型，A组在造模后第14天经尾静脉注射外源性的BMSCs1×107/ml，200ul；B组在造模后第14天起隔日经尾静脉注射外源性的BMSCs1×107/ml，200ul。以上各组小鼠造模后28天统一称重后处死，取小鼠全肺组织称重并留取部分肺组织行HE、MASSON染色及羟脯氨酸含量测定，用荧光定量PCR及免疫组化检测肺组织中SP-C和AQP5表达情况；提取肺组织的DNA检测雄鼠的性别决定基因(Sry基因)。 结果：1、肺系数： PC组及A、B组肺系数均较NC组高，PC组、A组、B组3组间无明显差异。2、肺组织病理：PC组及A组、B组肺泡炎、肺纤维化程度均较NC组明显加重；A组肺泡炎及纤维化较PC、B组稍减轻，但差异无统计学意义。3、HYP含量：PC组及A组、B组3组HYP含量均较NC组明显升高，A组较PC组、B组有所减低，与B组间的差异较为明显，有统计学意义，PC组、B组间无明显差异。4、Sry基因检测： A组、B组均有雄性小鼠的sry基因存在。5、肺组织SP-C蛋白的表达：PC组、A组、B组中SP-C蛋白的表达均低于NC组，A组的表达高于PC组、B组，而PC组与B组间无明显差异。6、肺组织AQP-5蛋白的表达：PC组、A组、B组的SP-C mRNA表达均低于NC组，PC组、A组、B组3组间无明显差异。7、肺组织SP-C mRNA表达情况：PC组、A组、B组3组中SP-C mRNA表达均低于NC组，，A组的表达高于PC组、B组，差异有统计学意义，而PC组与B组间无明显差异。8、肺组织AQP5mRNA表达情况：PC组、A组、B组的AQP5mRNA表达均低于NC组，3组间无明显差异。 结论：1、外源性BMSCs经尾静脉注射后可以到达博来霉素诱导的肺纤维化形成期小鼠的肺组织并可成功实现在局部定植；2、外源性BMSCs在肺纤维化形成期单次给药较单纯造模组的肺纤维化程度有减轻的趋势，但差异无明显统计学意义，因此目前尚不能说明在肺纤维化形成期给予外源性BMSCs干预能治疗肺纤维化；3、在肺纤维化形成期单次和多次给予外源性BMSCs的干预效果无统计学差异，甚至多次给药有加重肺纤维化的趋势。
[Abstract]:Aim: to investigate the effects of transplantation of exogenous bone marrow mesenchymal stem cells (BMSCs) on bleomycin-induced late pulmonary fibrosis in mice. Methods: the whole bone marrow adherent cells were isolated and cultured in male 6-8 week old ICR mice BMSCs. In this experiment, four groups were designed for 160 experimental mice. These mice were randomly divided into two groups: negative control group (NC group), positive control group (PC group) BMSCs intervention group (A (A group). The pulmonary fibrosis model of mice was made by bleomycin. Group A was injected with exogenous BMSCs 1 脳 107 / ml / ml 200ul1 脳 10 ~ (7) / ml on the 14th day after the establishment of the model. In group B, exogenous BMSCs 1 脳 107 / ml / ml was injected via caudal vein on the 14th day after modeling. All the above groups were killed after 28 days of unified weighing. The whole lung tissue of the mice was weighed and some of the lung tissues were taken for determination of Hedman son staining and hydroxyproline content. The expression of SP-C and AQP5 in lung tissue was detected by fluorescence quantitative PCR and immunohistochemistry. The male sex determinant gene (Sry gene) was detected by extracting DNA from lung tissue. Results compared with NC group, there was no significant difference in lung coefficient between PC group and Agna B group. There was no significant difference between group A and group B. The lung pathological changes of PC group and group A group were significantly worse than those of NC group. The pulmonary fibrosis degree in group A and group A was significantly higher than that in group C. The pulmonary alveolitis and fibrosis in group A were slightly less than those in group PCB, but there was no significant difference in HYP content between group A and group B, but the content of HYP in group A was significantly higher than that in group C, and the difference between group A and group B was more obvious than that in group B, but the content of HYP in group A was significantly higher than that in group C. There was no significant difference in the expression of SP-C gene between group B and group A: the expression of SP-C protein in lung tissue of group A was lower than that of group A (group A). The expression of SP-C protein in group B was lower than that in group A (group A), and the expression of SP-C protein in group B was lower than that in group A (group A). The expression of SP-C protein in lung tissue in group A was lower than that in group B. The expression of the protein in PC group was higher than that in PC group B group. However, there was no significant difference between PC group and B group. The expression of AQP-5 protein in lung tissue of PC group was significantly lower than that of group A and B group (P < 0.05). The expression of SP-C mRNA in lung tissue of PC group was significantly lower than that of group A and B group (P < 0.05). The expression of SP-C mRNA in lung tissue of PC group was significantly lower than that of group A and B group (P < 0.05). The expression of SP-C mRNA in three groups was lower than that in group A of NC group, and the expression of SP-C mRNA in group A was higher than that in group B of PC group. There was no significant difference in AQP5 mRNA expression between PC group and B group. The expression of AQP5 mRNA in lung tissue of group A was significantly lower than that of group B (P < 0.05). There was no significant difference in AQP5 mRNA expression between group B and group C (P < 0.05). ConclusionThe exogenous BMSCs can reach the lung tissue of bleomycin induced pulmonary fibrosis mice after injection through tail vein and can be successfully colonized locally. 2. The degree of pulmonary fibrosis of exogenous BMSCs at the stage of pulmonary fibrosis formation was alleviated, but the difference was not statistically significant. Therefore, at present, it is not clear that exogenous BMSCs can be used to treat pulmonary fibrosis in the stage of pulmonary fibrosis formation, but there is no statistical difference between single and multiple intervention effects of exogenous BMSCs at the stage of pulmonary fibrosis formation. Even multiple doses of drugs have a tendency to exacerbate pulmonary fibrosis.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R563.9
[Abstract]:Aim: to investigate the effects of transplantation of exogenous bone marrow mesenchymal stem cells (BMSCs) on bleomycin-induced late pulmonary fibrosis in mice. Methods: the whole bone marrow adherent cells were isolated and cultured in male 6-8 week old ICR mice BMSCs. In this experiment, four groups were designed for 160 experimental mice. These mice were randomly divided into two groups: negative control group (NC group), positive control group (PC group) BMSCs intervention group (A (A group). The pulmonary fibrosis model of mice was made by bleomycin. Group A was injected with exogenous BMSCs 1 脳 107 / ml / ml 200ul1 脳 10 ~ (7) / ml on the 14th day after the establishment of the model. In group B, exogenous BMSCs 1 脳 107 / ml / ml was injected via caudal vein on the 14th day after modeling. All the above groups were killed after 28 days of unified weighing. The whole lung tissue of the mice was weighed and some of the lung tissues were taken for determination of Hedman son staining and hydroxyproline content. The expression of SP-C and AQP5 in lung tissue was detected by fluorescence quantitative PCR and immunohistochemistry. The male sex determinant gene (Sry gene) was detected by extracting DNA from lung tissue. Results compared with NC group, there was no significant difference in lung coefficient between PC group and Agna B group. There was no significant difference between group A and group B. The lung pathological changes of PC group and group A group were significantly worse than those of NC group. The pulmonary fibrosis degree in group A and group A was significantly higher than that in group C. The pulmonary alveolitis and fibrosis in group A were slightly less than those in group PCB, but there was no significant difference in HYP content between group A and group B, but the content of HYP in group A was significantly higher than that in group C, and the difference between group A and group B was more obvious than that in group B, but the content of HYP in group A was significantly higher than that in group C. There was no significant difference in the expression of SP-C gene between group B and group A: the expression of SP-C protein in lung tissue of group A was lower than that of group A (group A). The expression of SP-C protein in group B was lower than that in group A (group A), and the expression of SP-C protein in group B was lower than that in group A (group A). The expression of SP-C protein in lung tissue in group A was lower than that in group B. The expression of the protein in PC group was higher than that in PC group B group. However, there was no significant difference between PC group and B group. The expression of AQP-5 protein in lung tissue of PC group was significantly lower than that of group A and B group (P < 0.05). The expression of SP-C mRNA in lung tissue of PC group was significantly lower than that of group A and B group (P < 0.05). The expression of SP-C mRNA in lung tissue of PC group was significantly lower than that of group A and B group (P < 0.05). The expression of SP-C mRNA in three groups was lower than that in group A of NC group, and the expression of SP-C mRNA in group A was higher than that in group B of PC group. There was no significant difference in AQP5 mRNA expression between PC group and B group. The expression of AQP5 mRNA in lung tissue of group A was significantly lower than that of group B (P < 0.05). There was no significant difference in AQP5 mRNA expression between group B and group C (P < 0.05). ConclusionThe exogenous BMSCs can reach the lung tissue of bleomycin induced pulmonary fibrosis mice after injection through tail vein and can be successfully colonized locally. 2. The degree of pulmonary fibrosis of exogenous BMSCs at the stage of pulmonary fibrosis formation was alleviated, but the difference was not statistically significant. Therefore, at present, it is not clear that exogenous BMSCs can be used to treat pulmonary fibrosis in the stage of pulmonary fibrosis formation, but there is no statistical difference between single and multiple intervention effects of exogenous BMSCs at the stage of pulmonary fibrosis formation. Even multiple doses of drugs have a tendency to exacerbate pulmonary fibrosis.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R563.9
【参考文献】
相关期刊论文 前8条
1 陈寅;马南;梅举;肖海波;陆善伟;徐怀阳;钟z
本文编号：2120865
本文链接：https://www.wllwen.com/yixuelunwen/huxijib/2120865.html
最近更新
教材专著
