1、The Role And Mechanism Of LncRNA AL110200 In Inflammation Process Of Atherosclerosis 2、Association Of Kallikrein Gene Polymorphism With Sporadic Intracranial Aneurysms In The Chinese Population

Objective:Atherosclerosis is recognized as a chronic inflammatory disease caused by synergy among multifactors that involve abnormal lipids metabolism, inflammation and dysfunction of immunity system and genetic factors, which leads to kinds of cardiovascular disease. The pathological processes of atherosclerosis involve in lipid deposition in vascular intima, endothelial cell damage, adhesion and infiltration of platelets and monocytes combined with hyperplasia of smooth muscle cells, the formation of foam cells. Among these processes, inflammation caused by monocytes migration to vascular intima is the first step in atherogenesis, which is important target of prevention of atherosclerosis. This process is regulated by many genes, including ALOX5 and PPARfβ/δ, which are associated with inflammation of atherosclerosis. Based on analysis results of bioinformation, we presumed that lncRNA AL110200 may regulate the expression of ALOX5 or PPARβ/δ. In our study, we aimed to explore the role and mechanism that maybe involved in the inflammation of atherosclerosis.Methods and Redults:To determine the potential role of lncRNA AL 110200 in inflammation of atherosclerosis, we performed QRT-PCR analysis to examine the expression of lncRNA AL110200 in THP-1 cells treated with LPS or oxLDL. Increased expression levels of lncRNA AL110200 were observed in THP-1 cells under LPS and oxLDL(1.85-fold, P<0.001 and 2.38-fold, P<0.01, respectively).And then we test the abilities of adhesion and migration of monocytes with ECM, we find the reducing expression of lncRNA AL 110200 prevent adhesion and migration of monocytes(0.65-fold and 0.55-fold, respectively; P<0.001).Moreover, we use qRT-PCR and ELISA to test the expression of cell factors associated with inflammation(such as IL-6, TNF-α and MCP-1) in THP-1 cells with down-regulation and up-regulation of lncRNA AL110200. The reducing (0.36-fold, 0.40-fold and 0.32-fold, respectively; P<0.001) and rising (1.75-fold,2.06-fold and 1.18-fold, respectively; P<0.001, P<0.001 and P<0.05, respectively) expressions of cell factors (IL-6, TNF-α and MCP-1) were obeserved in THP-1 cells with low expression and high expression of lncRNA AL110200, respectively. Furthermore, the results of ELISA were according to the change of mRNA.To determine the mechanism of pro-inflammation of lncRNA AL110200, we test the expressions of target genes after knock-down and over-expression of lncRNA AL110200 in THP-1 cells. We found that high expression of lncRNA AL110200 up regulatesd ALOX5 expression(3.03-fold, P<0.001) and knocking down of this lncRNA have opposite effect(0.23-fold, P<0.001); but the altertation of expression of PPARβ/δ was not obeserved in THP-1 cells with the high expression of lncRNA AL110200.To confirm the tagert gene of lncRNA AL110200, we analyzed the effects resulted from transfection of plasmids with different alleles of conserved SNP (2842). This SNP has conserved allele gene (T) across 14 species, the other allele gene is G. We found that transfection of the lncRNA AL110200 plasmid carrying SNP 2842 allele G led to reducing expression of ALOX5(0.46-fold, P<0.001). Accordingly, the over-expression of lncRNA AL110200 with this plasmid inhibits expressions of cell factors(IL-6, TNF-α and MCP-1) (0.11-fold,0.39-fold and 0.20-fold, respectively; P<0.001).Finally, the reduced expression levels of cytokines (TNF-α, IL-6 and MCP-1) were observed in THP-1 cells with knock-down of ALOX5 under the condition of overexpression of lncRNA AL110200. These results indicate that the role of lncRNA AL110200 in inflammation of atherosclerosis is achieved by regulating the expression of ALOX5. Conclusions:Our results support that the lncRNA AL110200 promote inflammation of atherosclerosis by mediating expression of ALOX5. These results may provide new clues to the mechanism and prevention of AS.Obeject:Intracranial aneurysm (IA) is a complex disease caused by synergy between genetic and environmental factors, with a high incidence; furthmore, the spontaneous subarachnoid hemorrhage (SAH) with ruptured IA has a high mortality rate (up to 50%), and poor outcome (20% patients with permanent activity). So understanding the pathological mechanism of IA is the best way to prevent and treat SAH patients. Many evidences support the important role of genetic factors in IA, so that we explore association study to identify candicate gene of IAs. Some studied report the function of Kallikreins in ECM remodeling, which may affect the formation and rupture of IA. And some GWAS and SNP studies find variants of Kallikreins was risk factors for intracranial aneurysm (IA) in a Finnish population, but these associations have not been test in Chinese population. In our study, we investigated the correlation between polymorphisms in the Kallikrein gene cluster with IA in Chinese population.Methods and Results:A multi-center case-control study was used to investigate the association Kallikrein variants with IA susceptibility in 308 cases and 443 controls. The differences in allelic frequencies between patients and the control group were evaluated with the chi-square test.After adjustment for traditional cardiovascular risk factors including medical and family history, smoking habits, and alcohol consumption, Kallikrein variant rs2808630 C alleles were significantlydecreased in case(odds ratio [OR],0.71; 95% confidence interval [CI],0.53-0.95; P=0.023). However, no association of the variant rs1701946 with sporadic IA was found (OR,0.78; 95% CI,0.57-1.06; P=0.115).Conclusions:Based on the results of our study, the variant rs1722561 of Kallikreins might reduce the risk of sporadic IAs among individuals of Chinese Han ethnicity, indicating that Kallikreins may be implicated in the formation and rupture of IA. The controversial results between studies for different population mainly result from the genetic heterogeneity in different ethnic populations.