Detection of expression levels of miR-194-5p and MagTl in peripheral blood mononuclear cells of patients with chronic HBV infection and their significances / 吉林大学学报(医学版)

ABSTRACT

Objective: To detect the expression levels of miR-194-5p and magnesium transporter protein 1 (MagTl) in peripheral blood mononuclear cells (PBMCs) of the patients with chronic hepatitis B virus (HBV) infection, and to investigate their possible clinical significances. Methods: The clinical materials of 40 healthy subjects (healthy control group). 40 cases of asymptomatic hepatitis B carriers (HBV carrier group). 40 patients with mild-moderate chronic hepatitis B (mild-moderate CHB group) and 40 patients with severe chronic hepatitis B (severe CHB group) were collected. The peripheral blood of the subjects in various groups was collected and the PBMCs were isolated by Ficoll-Hypaque density gradient centrifugation. The expression levels of miR-194-5p and MagTl mRNA in PBMCs of the subjects in various groups were detected by RT-PCR assay, the expression levels of MagTl protein in PBMCs of the subjects in various groups were detected by Western blotting method, the levels of Mg in peripheral blood and PBMCs of the subjects in various groups were detected, and the expression levels of CD6+ T lymphocyte surface molecules programmed death receptor 1 (PD-1) and natural killer cell receptor 2 group D molecule (NKG2D) in PBMCs of the subjects in various groups were analyzed by flow cytometry. The miR-194-5p mimic (miR-194-5p mimic group) and miR-jNC (miR-NC group) were transfected into the 293T cells, and then the dual luciferase reporter gene assay was used to detect the luciferase activities in 293T cells in two groups. Results: Compared with healthy control group, the expression levels of MagTl mRNA and protein, the expression levels of CD8 + T lymphocyte surface molecule NKG2D and the levels of Mg in PBMCs of the patients in HBV carrier group, mild-moderate CHB group and severe CHB group were decreased ( P<-0. 05) . the expression levels of miR-194-5p mRNA in PBMCs and CD8 + T lymphocyte surface molecule PD-1 of the patients were increased (P<.0. 05). Compared with HBV carrier group and mild-moderate CHB group, the expression levels of MagTl mRNA and protein, the expression level of CD8+ T lymphocyte surface molecule NKG2D and the level of Mg in PBMCs of the patients in severe CHB group were decreased ( P<0. 05). and the expression levels of miR-194-5p mRNA in PBMCs and CD8 + T lymphocyte surface molecule PD-1 of the patients were increased (P<0. 05). Compared with HBV carrier group, the expression levels of MagTl mRNA and protein, the expression level of CD8+ T lymphocyte surface molecule NKG2D and the level of Mg in PBMCs of the patients in mild-moderate CHB group were decreased ( P mRNA in PBMCs and CD6+ T lymphocyte surface molecule PD-1 of the patients were increased (P<0. 05). The dual luciferase reporter gene assay results confirmed that MagTl was a miR-194-5p target gene.

Conclusion:

MiR-194-5p may lead to chronic development of hepatitis B by down-regulating the expression of MagTl. causing MagTl-media ted Mg influx disorder and resulting in the decrease of the immune activity of CD8+ T lymphocyte.