Moxibustion regulates T-regulatory/T-helper 17 cell balance by modulating the microRNA-221/suppressor of cytokine signaling 3 axis in a mouse model of rheumatoid arthritis / 中西医结合学报

OBJECTIVE@#Rheumatoid arthritis (RA) progression is associated with the balance of T-regulatory (Treg) and T-helper 17 (Th17) cells, while the role of microRNAs (miRs) in regulating Treg/Th17 cell balance has not been clarified. This study aimed to assess whether moxibustion could regulate Treg/Th17 cell balance by modulating the miR-221/suppressor of cytokine signaling 3 (SOCS3) axis in the RA mouse model.@*METHODS@#A mouse model of collagen-induced arthritis (CIA) was established in male DBA/1J mice. Twenty-two days after CIA induction, the mice received daily treatment with moxibustion for 12 times. Pathological scores were assessed according to the levels of synovial hyperplasia. The expression levels of cytokines interleukin (IL)-1β, IL-6, tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), IL-17 and IL-10 were analyzed in serum by enzyme-linked immunosorbent assay. The cluster of differentiation 4 (CD4+) splenocytes was analyzed by fluorescence-activated cell sorting. The expression levels of RA-related miRs and target genes were subsequently detected, and the target of miR-221 was confirmed by the dual-luciferase reporter assay.@*RESULTS@#It was revealed that moxibustion treatment decreased the pathological scores and downregulated the expression levels of IL-1β, IL-6, TNF-α, IFN-γ and IL-17, while upregulated the expression level of IL-10. The Treg/Th17 cell balance was regulated by moxibustion treatment. The expression level of miR-221 was suppressed by moxibustion treatment. Furthermore, SOCS3 was found as the direct target of miR-221, which mediated the function of moxibustion by regulating the Treg/Th17 cell balance.@*CONCLUSION@#Moxibustion therapy regulated the Treg/Th17 cell balance by modulating the miR-221/SOCS3 axis in the RA mouse model.

Changes in expression of microRNA-221 and phosphatase and tension protein homologue in nerve stump after peripheral nerve injury / 中国修复重建外科杂志

Objective: To study the expressions of microRNA-221 (miR-221) and the protein of phosphatase and tension protein homologue (PTEN) in the proximal and distal stumps after sciatic nerve injury in rats and their correlation with the repair of peripheral nerve injury, so as to provide a new target for clinical diagnosis of peripheral nerve injury. Methods: Ninety-six male Sprague-Dawley rats of SPF grade were selected to establish sciatic nerve injury models. Twenty-four rats were sacrificed at 0 (immediately after operation), 1, 4, and 7 days after operation. The proximal and distal sciatic nerve fragments were taken under aseptic conditions. The expression of miR-221 was detected by real-time fluorescent quantitative PCR, and the expression of PTEN protein was detected by Western blot and immunofluorescent staining. The relationship between miR-221 and PTEN was verified by dual-luciferase reporter gene. At the same time, the ultrastructure of nerve stump was observed by transmission electron microscopy. Results: The results of real-time fluorescent quantitative PCR, Western blot, and immunofluorescence staining showed that the relative expression of miR-221 in the proximal and distal stumps increased gradually with time, and the relative expression of PTEN protein decreased gradually, and the differences between different time points after operation were significant ( P<0.05). At 1, 4, and 7 days after operation, the relative expression of miR-221 in proximal stump was significantly higher than that in distal stump, and the relative expression of PTEN protein in proximal stump was significantly lower than that in distal stump ( P<0.05). Dual-luciferase reporter gene suggested that PTEN was the target for miR-221. Transmission electron microscopy observation showed that the normal morphological structure was observed at 0 day after operation, and the proliferation of Schwann cells and degeneration of axons and myelin sheaths gradually increased with time. There was no significant difference between proximal and distal stumps at 1 day after operation. At 4 and 7 days, Schwann cells proliferated more in proximal stump than in distal stump, and the degeneration of axons and myelin sheaths was less. Conclusion: After sciatic nerve injury in rats, the up-regulation of the miR-221 expression targets the down-regulation of PTEN expression, which results in the difference of expression levels of miR-221 and PTEN in proximal and distal stumps. This phenomenon may play a role in promoting nerve repair after peripheral nerve injury.

MicroRNA-451a, microRNA-34a-5p, and microRNA-221-3p as predictors of response to antidepressant treatment

Aberrant expression of microRNAs (miRNAs) has been shown to be involved in early observations of depression. The aim of this study was to determine if serum levels of miRNA-451a, miRNA-34a-5p, and miRNA-221-3p can serve as indicators of disease progression or therapeutic efficacy in depression. We collected data from 84 depressed patients and 78 control volunteers recruited from the medical staff at the West China Hospital. Depression severity was rated using the 24-item Hamilton Depression Scale (HAMD). Serum miRNA-451a, miRNA-34a-5p, and miRNA-221-3p levels were determined in samples from the depressed patients before and 8 weeks after antidepressant treatment as well as in samples from controls. Compared with the controls, the patients had lower miRNA-451a levels, higher miRNA-34a-5p and miRNA-221-3p levels, and increased HAMD scores whether they underwent antidepressant treatment or not. Eight weeks after antidepressant treatment, the patients exhibited increased miRNA-451a levels, decreased miRNA-34a-5p and miRNA-221-3p levels, and reduced HAMD scores. The serum level of miRNA-451a was negatively correlated with HAMD scores of the patients, while the serum levels of miRNA-34a-5p and miRNA-221-3p were positively correlated with HAMD scores whether the patients underwent antidepressant treatment or not. Paroxetine was markedly effective in 50 patients who also displayed an increased level of miRNA-451a but reduced levels of miRNA-34a-5p and miRNA-221-3p. In contrast, paroxetine was moderately effective or ineffective in 34 patients. In conclusion, depressed patients had lower serum miRNA-451a but higher serum miRNA-34a-5p and miRNA-221-3p, and these miRNAs are potential predictors of the efficacy of antidepressants.

MicroRNA-221 promotes cell proliferation, migration, and differentiation by regulation of ZFPM2 in osteoblasts

Bone fracture is a common medical condition, which may occur due to traumatic injury or disease-related conditions. Evidence suggests that microRNAs (miRNAs) can regulate osteoblast differentiation and function. In this study, we explored the effects and mechanism of miR-221 on the growth and migration of osteoblasts using MC3T3-E1 cells. The expression levels of miR-221 in the different groups were measured by qRT-PCR. Then, miR-221 mimic and inhibitor were transfected into MC3T3-E1 cells, and cell viability and migration were measured using the CCK-8 assay and the Transwell migration assay. Additionally, the expression levels of differentiation-related factors (Runx2 and Ocn) and ZFPM2 were measured by qRT-PCR. Western blot was used to measure the expression of cell cycle-related proteins, epithelial-mesenchymal transition (EMT)-related proteins, ZFPM2, and Wnt/Notch, and Smad signaling pathway proteins. miR-221 was significantly up-regulated in the patients with lumbar compression fracture (LCM) and trochanteric fracture (TF). miR-221 promoted ALP, Runx2, and OPN expressions in MC3T3-E1 cells. miR-221 overexpression significantly increased cell proliferation, migration, differentiation, and matrix mineralization, whereas suppression of miR-221 reversed these effects. Additionally, the results displayed that ZFPM2 was a direct target gene of miR-221, and overexpression of ZFPM2 reversed the promoting effects of miR-221 overexpression on osteoblasts. Mechanistic study revealed that overexpression of miR-221 inactivated the Wnt/Notch and Smad signaling pathways by regulating ZFPM2 expression. We drew the conclusions that miR-221 overexpression promoted osteoblast proliferation, migration, and differentiation by regulation of ZFPM2 expression and deactivating the Wnt/Notch and Smad signaling pathways.

miR-221 promotes proliferation of lung cancer A549 cells by down-regu-lating PTEN / 中国病理生理杂志

AIM:To explore the effect of microRNA-221 (miR-221) on the proliferation of lung cancer A549 cells, and to investigate its mechanism .METHODS: The A549 cells were transfected with miR-221 mimics by Lipo-fectamine 2000.The expression of miR-221 was detected by RT-qPCR.The expression of PTEN at mRNA and protein le-vels was detected by RT-qPCR and Western blot , respectively .The cell proliferation was examined by CCK-8 assay and colony formation assay.The 3'-UTR of PTEN was cloned into luciferase reporter vector and its enzymatic activity was detec-ted to verify whether miR-221 targeted to PTEN.RESULTS:The expression level of miR-221 in the A549 cells was sig-nificantly increased after transfection with miR-221 mimics as compared with negative control group and blank group ( P<0.01).The mRNA and protein levels of PTEN were significantly down-regulated compared with control group and blank group ( P <0.05 ) .In addition , miR-221 over-expression significantly promoted the proliferation of A 549 cells ( P <0.05).Moreover, miR-221 inhibited the enzymatic activity of luciferase reporter vector of PTEN.CONCLUSION:Over-expression of miR-221 significantly promotes the proliferation ability of human lung cancer A 549 cells by down-regulation of PTEN.