miR-320c Regulates SERPINA1 Expression and Is Induced in Patients With Pulmonary Disease

IntroductionAlpha-1 antitrypsin deficiency (AATD) is a genetic condition resulting in lung and liver disease with a great clinical variability. MicroRNAs have been identified as disease modifiers; therefore miRNA deregulation could play an important role in disease heterogeneity. Members of miR-320 family are involved in regulating of multiple processes including inflammation, and have potential specific binding sites in the 3′UTR region of SERPINA1 gene. In this study we explore the involvement of miR-320c, a member of this family, in this disease.MethodsFirstly in vitro studies were carried out to demonstrate regulation of SERPINA1 gene by miR-320. Furthermore, the expression of miR-320c was analyzed in the blood of 98 individuals with different AAT serum levels by using quantitative PCR and expression was correlated to clinical parameters of the patients. Finally, HL60 cells were used to analyze induction of miR-320c in inflammatory conditions.ResultsOverexpression of miR-320 members in human HepG2 cells led to inhibition of SERPINA1 expression. Analysis of miR-320c expression in patient's samples revealed significantly increased expression of miR-320c in individuals with pulmonary disease. Additionally, HL60 cells treated with the pro-inflammatory factor lipopolysaccharide (LPS) showed increase in miR-320c expression, suggesting that miR-320c responds to inflammation.ConclusionOur findings demonstrate that miR-320c inhibits SERPINA1 expression in a hepatic cell line and its levels in blood are associated with lung disease in a cohort of patients with different AAT serum levels. These results suggest that miR-320c can play a role in AAT regulation and could be a biomarker of inflammatory processes in pulmonary diseases.     

软骨组织工程中种子细胞接种密度和比例的研究进展

目的综述软骨组织工程中种子细胞接种密度和比例的研究进展。方法广泛查阅三维结构中构建组织工程软骨的相关文献，对最常用的种子细胞接种密度与比例进行综述。结果关节软骨细胞（articular chondrocytes，ACHs）和BMSCs是软骨组织工程中最常用的种子细胞，体内、外实验显示两种细胞均具有一定形成透明软骨的能力。而种子细胞的接种密度和比例对最终形成的组织工程软骨质量有重要影响。ACHs或BMSCs细胞接种密度接近或高于正常关节软骨中的细胞密度时，可构建出质量较好的组织工程软骨。在相同培养条件下，BMSCs在单独应用时形成透明软骨的能力不如ACHs或两者共培养。结论受支架材料、生长因子、细胞代次等因素影响，软骨组织工程中最优的细胞接种密度与比例仍需进一步研究。