摘要
目的:筛选与高血压病血瘀证相关微小RNA(microRNA,miRNA),从基因组学探索高血压病血瘀证的发病机制。方法:运用高血压病气虚血瘀证、气滞血瘀证、寒凝血瘀证、热结血瘀证、非血瘀证患者和健康人的血清干预人脐静脉血管内皮细胞CRL-1730,建立高血压病血瘀证血管内皮细胞损伤模型;提取各组总RNA,采用Solexa高通量测序法和数字基因表达谱测序原理对各组样本进行测序分析,筛选各组间的差异表达miRNA和mRNA,使用靶基因miRWalk预测软件对2者进行相关性的整合,筛选与血瘀证相关miRNA,qRT-PCR定量分析验证miRNA的表达。结果:整合气虚血瘀组与非血瘀组、气滞血瘀组与非血瘀组、寒凝血瘀组与非血瘀组、热结血瘀组与非血瘀组相关数据后,在高表达的基因群中发现SAT1-Hsa-miR-199a-5p和ATF4-Hsa-miR-1283;qRT-PCR定量检测Hsa-miR-199a-5p和Hsa-miR-1283在高血压病血瘀证组和非血瘀证组中的上调与下调趋势与基因测序结果一致。结论:Hsa-miR-199a-5p和Hsa-miR-1283可能是高血压病血瘀证形成的特异性相关miRNA。
AIM：To screen the miRNA related to blood stasis syndrome and to explore the genetic mechanism of blood stasis syndrome in hypertension.METHODS：Human umbilical vein endothelial cell line CRL-1730 was co-cul-tured with the human sera from healthy normal controls and hypertension patients to establish a blood stasis-hypertension cell modela.The hypertension patients showed qi deficiency and blood stasis, qi stagnation and blood stasis, cold retaining and blood stasis, heat retaining and blood stasis and non-blood stasis syndromes.The endothelial cell models of blood stasis in hypertension were established.Target cells were collected for total RNA extraction.The techniques of Solexa （ high-throughput sequencing method） and digital gene expression profiling were applied for screening the target miRNA, and miRWalk software was used for online prediction the mRNA which might be the relevant target.qRT-PCR was conducted to confirm the prediction.RESULTS：SAT1-Hsa-miR-199a-5p and ATF4-Hsa-miR-1283 were found to be highly expressed in these cell models.The expression of miR-1283 and miR-199a-5p was confirmed by qRT-PCR.CONCLUSION：miR-199a-5p and miR-1283 may be the relevant miRNA for the prediction of blood stasis syndrome in hypertension.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2015年第5期817-822,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81173157)
广东省自然科学基金资助项目(No.10151063201000045)
关键词
高血压病
血瘀证
微小RNA
Hypertension
Blood stasis syndrome
MicroRNA
