Mechanism of Gentisic Acid on Rheumatoid Arthritis Based on miR-19b-3p/RAF1 Axis / 中国结合医学杂志

OBJECTIVE@#To investigate the therapeutic effect of gentisic acid (GA) on rheumatoid arthritis (RA) based on the miR-19b-3p/RAF1 axis.@*METHODS@#The cell counting kit-8 method was used to detect the growth inhibitory effect of different concentrations of GA on MH7A cells, and the drug concentration of GA was determined in the experiment. The quantificational real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-19b-3p and RAF1. RAF1, extracellular regulated protein kinases1/2 (ERK1/2) and phospho-ERK1/2 (p-ERK1/2) were examined by Western blotting. Three methods (dual-luciferase assay, qRT-PCR and Western blot analysis) were used to verify miR-19b-3p targeting RAF1. Flow cytometry was performed to detect MH7A cell apoptosis. Transwell and wound healing assays were used to determine the invasion and migration capacities of MH7A cells.@*RESULTS@#The growth of MH7A cells was gradually inhibited with increasing GA concentration. When the GA concentration exceeded 80 mmol/L, GA was significantly cytotoxic to MH7A cells, so the half maximal inhibitory concentration of GA for MH7A cells was calculated as 67.019 mmol/L. GA upregulated miR-19b-3p expression, downregulated RAF1 expression, inhibited ERK1/2 phosphorylation, induced MH7A cell apoptosis and suppressed MH7A cell invasion and migration (P<0.05 or P<0.01). RAF1 was identified as the target of miR-19b-3p and reversed inhibitory effects on miR-19b-3p expression (P<0.05 or P<0.01). The miR-19b-3p inhibitor upregulated RAF1 expression and ERK1/2 phosphorylation, suppressed MH7A cell apoptosis and induced MH7A cell invasion and migration (P<0.01).@*CONCLUSION@#GA regulated miR-19b-3p/RAF1 axis to mediate ERK pathway and inhibit the development of RA.

The relationship between differentially expressed miR-19b-3p and cognitive function in patients with Alzheimer's disease / 中华行为医学与脑科学杂志

Objective To explore the relationship between differentially expressed miR-19b-3p and cognitive function in patients with Alzheimer's disease (AD).Methods The miRNA expression profiles of patients with AD(AD group,n=30) or healthy elderly people (NC group,n=30) were analyzed by Illumina/Solexa high-throughput sequencing technique.The miRNA lentiviral plasmids were constructed and injected into the model of AD rats.The cognitive function of rats was analyzed with water maze test.The mimics and inhibitors were synthesized and transfected into SH-SY5Y cell lines.The protein expression of β-amyloid precursor protein cleavage enzyme 1 (BACE1) was detected by western blot.Results Compared with the NC group,the expression of miR-146a-5p (log2AD/Control =2.3),miR-195-5p (log2 AD/Control =10.2),miR-20b-5p(log2AD/Control =15.1) in serum of AD group was up-regulated and the expression of miR-19b-3p (log2 AD/Control =-8.0) and miR-125b-3p (log2 AD/Control =-15.3) was down-regulated (P＜ 0.05).Compared with the rats in the AD group((26.50±3.12) s),the rats in the AD+miR-19b-3p group ((15.33±2.78) s) showed significantly shorter escape latencies(P＜0.05).Compared with the NC group,the protein expression of BACE1 in miR-19b-3p mimetic group was decreased and the protein expression of BACE1 in inhibitor group was increased.Conclusion miR-19b-3p may improve the cognitive function of AD patients via regulating the expression of BACE1.