产前尼古丁暴露对成年期子代大鼠血管内皮功能和IGFs轴的影响

发布时间：2018-06-24 13:46

本文选题：程序性控制 + 血管内皮功能　；参考：《福建医科大学》2013年硕士论文

【摘要】：背景烟草暴露与心血管疾病有关联，尼古丁是烟草中最主要的致病成分。生命发育早期的环境与母体等因素可在胚胎期形成印痕，程序性控制出生后甚至成年期心血管疾病的发生。血管内皮功能损伤和胰岛素样生长因子(insulin likegrowth factors，IGFs)轴异常参与介导心血管疾病发生发展。目前还不清楚孕妇尼古丁暴露对日后成年期子代血管内皮功能和IGFs轴的影响。目的本研究旨在明确产前尼古丁暴露（Prenatal Nicotine Exposure, PNE）对子代大鼠血管内皮功能和IGFs轴的影响及其潜在的性别依赖性差异、并阐明这些效应是否可持续作用至日后成年期。方法孕SD大鼠20只随机分成两组：PNE组和正常对照组，均予皮下植入渗透微泵（2ML4型），其中PNE组渗透微泵携带尼古丁(8mg/kg/day)，正常对照组渗透微泵仅携带生理盐水。待子代大鼠满5月龄时，腹腔麻醉，迅速截取胸主动脉及肠系膜动脉各4mm，采用Power Lab数据采集分析系统测定血管内皮功能；收集血样，采用ELISA酶联免疫法测定血清IGF-1、 IGF-2、 IGFBP-1、IGFBP-2及IGFBP-3水平；取胸主动脉、心、肺、肝、肾、胰腺等组织标本，行HE染色及免疫组化染色检测IGF-1、IGF-1R、IGF-2、IGFBP-1、IGFBP-2及IGFBP-3的组织特异性表达水平。结果 1) Ach与SNP诱导的子代大鼠胸主动脉内皮依赖/非依赖性血管舒张百分数，PNE组与正常对照组之间无显著差异（均P＞0.05）； 2) Ach与SNP诱导的子代大鼠肠系膜动脉的内皮依赖/非依赖性血管舒张百分数， PNE组雄性子代较正常对照组显著降低（P＜0.05），而雌性子代较正常对照组显著升高（P＜0.05）； 3)子代大鼠胸主动脉组织HE染色可见管腔不平、内皮细胞空泡变性、内皮部分脱落、内膜增厚，中膜平滑肌明显萎缩、细胞排列紊乱，弹性纤维层结构不清；肺脏组织HE染色在小支气管周围肺泡间质间可见大量炎症细胞浸润、淋巴滤泡形成；肾脏组织HE染色可见肾小球周围小动脉扩张； 4) PNE组雌性子代大鼠血清IGFBP-3水平比正常对照组显著降低（P＜0.05）,但是雄性子代大鼠血清IGFBP-3水平与正常对照组无显著差异（P＞0.05）；此外，PNE组子代大鼠血清IGF-1、IGF-2、IGFBP-1和IGFBP-2水平均与正常对照组无显著差异（均P＞0.05）； 5)在肾脏组织，正常对照组雄性子代IGF-1、IGF-1R、IGF-2、IGFBP-1、IGFBP-2和IGFBP-3均呈较强表达（+++），而雌性子代则呈弱表达（±～+），PNE组雄性子代（+～++）较正常对照组表达下调，而雌性子代（+～++）则较正常对照组表达上调。在肝脏组织，正常对照组子代IGF-1、IGF-1R、IGF-2、IGFBP-1、IGFBP-2和IGFBP-3均呈弱表达（±～+），而PNE组子代（+～++）表达上调；在肺脏组织， PNE组子代IGF-1、IGF-1R、IGF-2、IGFBP-1、IGFBP-2和IGFBP-3于肺泡间质小动脉周围细胞胞浆有少量表达（±），而正常对照组未见表达；在胰腺、心脏以及胸主动脉等脏器组织，PNE组与正常对照组均未见表达。结论 1) PNE可影响5月龄子代大鼠肠系膜动脉内皮依赖/非依赖性血管舒张功能，该效应存在性别差异，提示PNE所致血管内皮功能损伤始于中型肌性动脉，可持续作用至成年期，且以雄性子代血管内皮功能损伤为特征； 2) PNE可导致5月龄子代大鼠胸主动脉血管内膜及中膜病理结构损害，提示PNE不但可引起血管功能损伤还可导致血管结构的病理改变；此外，还可导致肺脏小支气管周围肺泡间质间淋巴滤泡形成、炎症细胞浸润； 3) PNE可下调5月龄雌性子代大鼠血清IGFBP-3水平，并导致子代大鼠IGFs及IGFBPs组织特异性表达异常，，其中肾脏的表达水平存在性别差异，PNE对子代IGFs轴的调控可能参与介导PNE诱导的血管内皮功能损伤。
[Abstract]:Background Tobacco exposure is associated with cardiovascular disease , nicotine is the most important pathogenic component in tobacco . In the early stage of life development , the factors such as maternal and other factors can form marks on the embryonic stage . It is not clear that the vascular endothelial function injury and insulin - like growth factors ( IGFs ) axis are involved in the development of cardiovascular disease .

Objective To investigate the effects of prenatal nicotine exposure ( PNE ) on vascular endothelial function and IGFs axis in offspring rats and their potential gender - dependent differences .

Methods 20 SD rats were randomly divided into two groups : PNE group and normal control group , which were implanted subcutaneously with osmotic micro - pump ( 2ML4 type ) , in which PNE group osmotic micro - pump carries nicotine ( 8 mg / kg / day ) , normal control group osmotic micro - pump carries nicotine ( 8 mg / kg / day ) .
Blood samples were collected , and serum IGF - 1 , IGF - 2 , IGFBP3 were measured by ELISA - linked immunosorbent assay ( ELISA ) .
The tissue - specific expression level of IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 were detected by HE staining and immunohistochemical staining .

Results

1 ) There was no significant difference between the endothelium - dependent / non - dependent vasodilatation percentage of thoracic aorta and PNE group and normal control group ( P > 0.05 ) .

2 ) The endothelium - dependent / non - dependent vasodilatation percentage of mesenteric artery was significantly lower in PNE group than in normal control group ( P & lt ; 0.05 ) , while the female offspring was significantly higher than that of normal control group ( P & lt ; 0.05 ) .

3 ) HE staining of the thoracic aorta in the offspring rats showed that the lumen was not uneven , the vacuolar degeneration of the endothelial cells , the loss of endothelium , the thickening of the intima , the obvious atrophy of the smooth muscle of the medial membrane , the disorder of the cell arrangement , the structure of the elastic fibrous layer were not clear ;
HE staining of the lung tissue showed a large amount of inflammatory cell infiltration and lymphoid follicles formation between the alveolar spaces around the small bronchus .
The renal tissue HE staining showed the expansion of arterioles around the glomerulus ;

4 ) There was no significant difference ( P < 0 . 05 ) between the levels of serum - 3 - 3 in the serum of the female offspring of the PNE group ( P < 0 . 05 ) , but there was no significant difference between the levels of serum - 3 - 3 and the normal control group ( P > 0 . 05 ) .
In addition , the levels of serum IGF - 1 , IGF - 2 , IGFBP3 and IGF - 2 in the offspring of PNE group were not significantly different from those in the normal control group ( P > 0.05 ) .

In the normal control group , the expression of IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1R , IGF - 2 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 , IGFBP3 and IGF - 1 , IGF - 1R , IGF - 2 ,
There was a small expression of IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 in the pulmonary tissue , PNE group , IGF - 1 , IGF - 1R , IGF - 2 , IGFBP3 and IGFBP3 .
In the pancreas , heart and thoracic aorta , the PNE group and the normal control group were not expressed .

Conclusion

1 ) PNE can affect the endothelium - dependent / non - dependent vasodilatation function of mesenteric artery in 5 - month - old offspring rats , and the effect is gender difference , suggesting that the damage of vascular endothelial function caused by PNE starts from the medium - sized muscular artery , which can be continuously applied to adulthood , and is characterized by the injury of the endothelial function of the vascular endothelium in the male offspring ;

2 ) PNE can lead to the damage of vascular intima and pathological structure of thoracic aorta in 5 - month - old rats , suggesting that PNE not only can cause vascular function injury but also can lead to pathological changes of vascular structure ;
In addition , it can also lead to the formation of lymph follicles and infiltration of inflammatory cells in the interstitial interstitial spaces around the lungs of the lungs .

3 ) PNE could down - regulate the levels of serum levels of IGFs and IGFBPs in the offspring of 5 - month - old female rats , and lead to abnormal expression of IGFs and IGFBPs in offspring rats , in which the expression level of PNE is gender difference , and the regulation of PNE on children ' s IGFs may be involved in mediating the vascular endothelial function injury induced by PNE .
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R114

【参考文献】