Research Progress of microRNA-7/124/155 in Parkinson's Disease / 中国医学科学院学报

Parkinson's disease(PD)is the second most common neurodegenerative disease after Alzheimer's disease,with high morbidity and high disability rate.Since the early symptoms of PD are not typical and often similar to those of normal aging or other diseases.It is easy to missed diagnosis and misdiagnosis,which seriously affects the diagnosis and treatment of this disease and aggravetes the burden on the patients' life.MicroRNAs(miRNA)are a class of endogenous non-coding RNAs that are involved in post-transcriptional regulation by binding to target messenger RNAs(mRNA).They are highly conserved,short,easy to obtain,and can stably exist in peripheral body fluids.They have been used as biomarkers for a variety of diseases.Recent studies have demonstrated that miRNA play an important role in the development of PD.This paper reviews the recent research progress of miR-7/124/155,three mature miRNA in PD,aiming to provide reference for clarifying the pathogenesis and guiding the diagnosis and treatment of PD.

Effect of miR-124a on collagen-induced arthritis in mice and the underlying mechanisms / 中南大学学报(医学版)

OBJECTIVES@#Rheumatoid arthritis (RA) is a chronic autoimmune disease. MicroRNA has been shown to play an important role in RA. MicroRNA-124a (miR-124a) has anti-proliferative and anti-inflammatory effects in RA fibroblast synovial cells. This study aims to explore the effects of miR-124a overexpression on arthritis in collagen-induced arthritis (CIA) mice and the underlying mechanisms.@*METHODS@#Bovine type II collagen and complete Ferris adjuvant were used to induce CIA model from DBA/1 mice. Twenty-eight days after initial immunization (D28), CIA mice were randomly divided into a model group, a miR-124a treatment group, and a negative control (NC) group. Physiological saline, miR-124a agomir, and miR-124a agomir NC were injected into the skin at the tail root of mice every 3 days for 4 times, respectively. The degree of joint swelling and arthritis index of mice were recorded accordingly. Sixty-three days after initial immunization (D63), the mice were sacrificed to obtain the synovial tissue of ankle joint. HE staining was used to observe the proliferation of synovial cell, infiltration of inflammatory cell, pannus, and bone erosion of synovial tissues; TUNEL staining was used to detect cell apoptosis; qRT-PCR was used to detect the mRNA expression of miR-124a, phosphatidylinositol-3-kinase catalytic subunit alpha (PIK3CA) and its downstream genes Bcl-2 and Bax. Immunohistochemistry was used to detect the protein expression of PIK3CA, Bcl-2, and Bax protein in synovial tissues of each group.@*RESULTS@#Different degrees of swelling presented in the paws of DBA/1 mice at D28, which indicated the CIA model was constructed successfully. Forty-eight days after initial immunization (D48), the paws of mice in the miR-124a treatment group were only slightly red and swollen, while the paws of mice in the model group and the NC group were obviously red and swollen. The arthritis index of mice in the miR-124a treatment group were decreased significantly compared to the NC group at D51, D53, D59, and D62 (51, 53, 59, 62 days after initial immunization) (all P<0.05). Sixty-three days after initial immunization (D63), HE staining indicated that the scores of synovial cell proliferation, inflammatory cell infiltration, synovial pannus, and bone erosion were significantly reduced in the miR-124a treatment group (P<0.05 or P<0.01), while cell apoptosis was increased in the miR-124a treatment group compared with the model group and NC group (P<0.01 or P<0.001). Besides, the expression of miR-124a and Bax in the synovial tissue in miR-124a treatment group was significantly higher than those in the model group and NC group (P<0.01 or P<0.001), while the expressions of PIK3CA and Bcl-2 were decreased (P<0.05 or P<0.01 or P<0.001), and the ratio of Bcl-2 to Bax was significantly decreased (P<0.01 or P<0.001).@*CONCLUSIONS@#Overexpression of miR-124a can reduce arthritis in CIA mice bacause it could promote synovial cell apoptosis and inhibit synovial cell proliferation via targeting PIK3CA and regulating its downstream pathways.

Study on adsorption of microRNA-124 by long chain non-coding RNA MALAT1 regulates osteogenic differentiation of mesenchymal stem cells / 中国修复重建外科杂志

Objective: To investigate the regulatory effect of long chain non-coding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) adsorbing microRNA-124 (miR-124) on osteogenic differentiation of mesenchymal stem cells (MSCs). Methods: C3H10T1/2 cells derived from mouse embryos were cultured in vitro, then randomly divided into control group (group A), lncRNA MALAT1 no-load plasmid group (group B), lncRNA MALAT1 overexpression plasmid group (group C), lncRNA MALAT1 small interfering RNA (siRNA) group (group D), and lncRNA MALAT1 siRNA negative control group (group E). The cells were transfected into plasmids and siRNA, then induced to differentiate into osteoblasts. Alkaline phosphatase (ALP) and alizarin red staining were used to detect the osteogenic differentiation of cells in each group, real-time fluorescence quantitative (qRT-PCR) analysis was used to detect the expressions of lncRNA MALAT, miR-124, and osteogenesis-related genes such as Runt-related transcription factor 2 (Runx2), osteopontin (OPN), and osteocalcin (OCN) in each group. Double luciferase reporter gene was used to detect the targeting regulation of lncRNA MALAT1 to miR-124. Results: The relative contents of ALP positive cells, mineralized nodule, and the relative mRNA expressions of lncRNA MALAT1, Runx2, OPN, and OCN in group C were significantly higher than those in other groups ( P0.05). The results of double luciferase reporter gene assay showed that lncRNA MALAT1 targeting down-regulated the expression of miR-124. Conclusion: LncRNA MALAT1 can targeting down-regulate the expression of miR-124 and promote the osteogenic differentiation of MSCs.

Effect of gap junctions on anti-proliferation effect of miR-124 and its mechanism / 中国药理学通报

was observed by patch clamp. Results Cx32 or Cx26 expression and GJ function were induced by doxycycline (Dox, the promotor for PBI plasmid) in transfected Hela cells. MiR-124 reduced the proliferation of Hela cells, dox incubation alone did not affect Hela cell growth, and also had no effect on anti-tumor effect of miR-124 when combined with miR-124 transfection. Compare with U 87shRNA-Cx43 , the Cx43 expression and GJ function significantly decreased in U87shRNA-Cx43. Similar to the effect on Hela cells, MiR-124 also reduced U87 cell growth. Reducing Cx43 expression did not influence U87 cell proliferation, but attenuated the growth-inhibition effect of miR-124 when combined with miR-124 transfection. Under the microscope, the transfer of fluorescence-labelled miR-124 from "donor" cell to adjacent " non-injection " cell was observed. Conclusions The role of GJ on anti-tumor effect of miR-124 possesses connexin heterogeneity. Compare with Cx26 or Cx32, GJ composed of Cx43 has more obvious effect, which may be related to the maximum permeability of junction channel to miR-124.

Effects of miR-124a overexpression on expression levels of TNF-α and IL-6 and cell cycle of J774. 1 cells / 吉林大学学报(医学版)

Objective; To explore the effects of miR-124a on the expression levels of tumor necrosis factor alpha (TNF-a) and interleukin-6 (IL-6) and cell cycle in the macrophage J774. 1 cells of the rheumatoid arthritis (RA) model mice, and to elucidate the mechanism of miR-124a in the pathogenesis of RA. Methods: The J774. 1 cells in logarithmic growth phage were obtained and uniformly inoculated in the petri dish with 1 × 106 mL-1, and there were 3 multiple holes in each group; transfection was carried out when the fusion degree of the cells reached 60%. The cells were divided into blank control group (without any treatment), empty vector group (transfected with adenovirus negative fluid) and miR-124a overexpression group (transfected with miR-124a adenovirus vector). The expression levels of TNF-α and IL-6 in supernatant of the cells in three groups were detected by ELISA 48 h after transfection. RT-PCR was used to detect the relative expression levels of TNF-α and IL-6 mRNA in J774. 1 cells 48 h after transfection and the changes of cell cycle were detected using flow cytometry. Results: The expression levels of TNF-α and IL-6 in supernatant of the cells in miR-124a overexpression group were lower than those in blank control group and empty vector group 48 h after transfection (P=0. 038, P=0. 042; P=0. 043, P=0. 044). The expression levels of TNF-α mRNA and IL-6 mRNA in miR-124a group were significantly lower than those in blank control group and empty vector group (P = 0. 001, P=0.002; P=0. 001, P=0. 003). The pecentage of cells in Gi phase in miR-124a overexpression group was higher than those in blank control group and empty vector group (P<0. 01); the percentages of cells in S phase and G2 phase were lower than those in blank control group and empty vector group (P<0.01). Conclusion: MiR-124a can decrease the expression levels of inflammatory cytokines after transfecting the macrophages J774. 1 cells, and inhibit the proliferation of cells. MiR-124a is a potential therapeutic target for the treatment of RA.

A Study of microRNA-124 Expression in blood Serum of Patients with Intracerebral Hemorrhage / 昆明医科大学学报

Objective To observe microRNA-124 expression in the serum of patient with intracerebral hemorrhage and explore the relationship between the expression and associated clinical factors. Methods Thirty serum specimen were collected among patients with intracerebral hemorrhage, who were checked up at the same time in the hospital. Relative expression of microRNA-124 in serum was detected. The variation of MicroRNA-124 expression between the two groups and clinical data of patients with intracerebral hemorrhage were analyzed. The relationship between microRNA-124 and clinical factors related to intracerebral hemorrhage was explored. Results Compared with healthy people, microRNA- 124 expression in serum increased among patients with intracerebral hemorrhage (P＜0.05) . The expression is related to the bleeding volume and onset time (P＜0.05) . Meanwhile, no obvious correlation is established between serum microRNA-124 expression and other factors in patients with intracerebral hemorrhage, such as gender, bleeding area, with or without surgical treatment (including minimally invasive and craniotomy), a history of high blood pressure, fasting venous blood glucose levels (FPG), and cerebrovascular disease history (P＞0.05) . Conclusion The serum microRNA-124 expression in patients with intracerebral hemorrhage is higher than that in healthy people. Bleeding volume positively increases expression. A higher expression is seen within one week at the onset of intracerebral hemorrhage compared to that one week after.

Nicotine protects against DSS colitis through regulating microRNA-124 and STAT3 / 中国药理学与毒理学杂志

OBJECTIVE Although it is generally believed that nicotine accounts for the beneficial effect of smoking on ulcerative colitis, the underlying mechanisms remain not well understood. Our previous finding that nicotine inhibits inflammatory responses through inducing miRNA-124 prompted us to ask whether the miRNA is involved in the protective action of nicotine on UC. METHODS MiR-124 expres-sion in colon tissues and cells was determined by q-PCR and in situ hybridization.The effect of miR-124 on protective role of nicotine in ulcerative colitis was evaluated in DSS-treated mice and IL-6-treated Caco-2 colon epithelial cells. Expression of p-STAT3/STAT3 was detected by immunohistochemistry and Western-blot analysis. RESULTS miR-124 expression is upregulated in colon tissues from UC patients and DSS-induced colitis mice. Nicotine treatment further elevated miR-124 level in lympho-cytes isolated from human ulcerative colonic mucosa and ulcerative colon tissues from DSS mice,both in infiltrated lymphocytes and epithelial cells. Administration of nicotine also reduced weight loss, improved DAI and decreased HE score in DSS-induced colitis mice.Moreover,knockdown of miR-124 in vivo significantly diminished the beneficial effect of nicotine on murine colitis, and in vitro on IL-6-treated Caco-2 colon epithelial cells.Further analysis indicated that nicotine inhibited STAT3 activation in vivo and in IL-6-treated Caco-2 colon epithelial cells and Jurkat human T lymphocytes,in which miR-124 knockdown led to increased activation of STAT3. Blocking STAT3 activity alone is beneficial for DSS colitis and also abolished nicotine′s protective effect in this model.CONCLUSION These data indicated that nicotine exerts its protective action in UC through inducing miR-124 and its effect on STAT3, and suggest that the miR-124/STAT3 system is a potential target for the therapeutic intervention of UC.

Expression change of microRNA-124 and its correlation with axon regeneration after traumatic brain injury in mice / 中华创伤杂志

Objective To observe the expression changes of microRNA-124(miRNA-124) following traumatic brain injury (TBI) in mice and investigate the correlation of miRNA-124 with neural axon regeneration.Methods Ninety-one C57BL/6 mice were assigned into TBI group (n =63) and control group (n =28) according to the random number table.Mice in TBI group were subjected to controlled cortical impact and euthanized at 12 hours and 1,3,7,14,21,28 days postinjury for the collection of brain tissue in the trauma zone.Mice in control group underwent craniectomy only.Trauma zone observation was done using the HE staining.Expression of miRNA-124 was detected using the real-time PCR.Levels of Nrp-1,Gap-43 and Tau were detected using the Western blot and immunohistochemical staining.Results After injtury,study of mice behavior and HE staining indicated the establishment of experimental model was successful.Expression of miRNA-124 reached the peak at 3 days postinjury (3.80 ± 0.22),expression of Nrp-1 reached the peak at 7 days postinjury (2.006 ±0.179),expression of Tau reached the peak at 14 days postinjury (2.063 ±0.172),and expression of Gap-43 sustained high level since 12 hours after injury(1.355 ± 0.093) (P ＜ 0.05).Count of axon marker positive cells in TBI group was the lowest at 1 day postinjury due to the direct damage and edema,and then slowly recovered.There was no significant difference in the count of axon marker positive cells between the two groups at 14,21 and 28 days postinjury (P ＞ 0.05),but the morphology in TBI group changed obviously.Although the positive cells of axon marker decreased at 1 day postinjury,expressions of miRNA-124,Nrp-1,Tau and Gap-43 in TBI group were significantly increased compared to the detections in control group (P ＜ 0.05).Conclusion Increased expression of miRNA-124 in trauma zone may closely related to axon regeneration after TBI in mice.

Nicotine protects against ulcerative colitis through regulating microRNA-124 and STAT3 / 中国药理学与毒理学杂志

OBJECTIVE Although it is generally believed that nicotine accounts for the beneficial effect of smoking on ulcerative colitis, the underlying mechanisms remain not well- understood. Our previous finding that nicotine inhibits inflammatory responses through inducing miRNA-124 prompted us to ask whether the miRNA is involved in the protective action of nicotine on UC. METHODS MiR-124 expression in colon tissues and cells was determined by q-PCR and in situ hybridization. The effect of miR-124 on protective role of nicotine in ulcerative colitis was evaluated in DSS-treated mice and IL-6-treated Caco-2 colon epithelial cells. Expression of p-STAT3/STAT3 was detected by immuno?histochemistry and Western blot analysis. RESULTS miR- 124 expression is upregulated in colon tissues from patients and DSS- induced colitis. Nicotine treatment further elevated miR- 124 level in colon tissues of the mice, in infiltrated lymphocytes and epithelial cells, and augmented miR- 124 expression in lymphocytes isolated from human ulcerative colon tissues. Administration of nicotine also reduced weight loss, improved DAI and decreased HE score in DSS-induced colitis. Moreover, knock?down of miR-124 in vivo significantly diminished the beneficial effect of nicotine, and in vitro on IL-6-treated Caco-2 colon epithelial cells. Further analysis indicated that nicotine inhibited STAT3 activation in vivo and in IL-6-treated Caco-2 colon epithelial cells and Jurkat human T lymphocytes, in which miR-124 knockdown led to increased activation of STAT3. CONCLUSION These data indicated that nicotine exerts its protective action in UC through inducing miR-124 and its effect on STAT3, suggesting that the miR-124/STAT3 system is a potential target for the therapeutic intervention of UC.

Effects of acupuncture at Baihui (GV 20) and Zusanli (ST 36) on peripheral serum expression of MicroRNA 124, laminin and integrin β1 in rats with cerebral ischemia reperfusion injury / 中国结合医学杂志

<p><b>OBJECTIVE</b>To explore the effects of acupuncture at Baihui (GV 20) and Zusanli (ST 36) on the peripheral serum expression of microRNA 124 (miRNA 124), laminin and integrin β1 in rats with cerebral ischemia reperfusion injury (CIRI).</p><p><b>METHODS</b>Seventy-two healthy male Sprague-Dawley rats were randomized into a model group, an acupuncture group, and a sham-operated group using a random digits table, with 24 rats per group. Each group was further randomly divided into 1-, 3-, 5-, and 7-day subgroups based on the reperfusion time according to a random digits table, with 6 rats in each subgroup. In the model and acupuncture groups, CIRI was induced using the thread occlusion method. Electroacupuncture stimulation was applied daily to GV 20 and left ST 36 for 20 min at the indicated time points after successful operations. Serum was sampled for detecting laminin and integrin β1 protein via enzyme-linked immunosorbent assay, and serum miRNA 124 was examined using quantitative polymerase chain reaction.</p><p><b>RESULTS</b>The serum level of miRNA 124 in the cerebral ischemia rats increased significantly, and the peak expression of miRNA 124 in both the model and acupuncture groups occurred at 3 days. The expression of miRNA 124 in the acupuncture group was higher than in the model group at the same time point (5.96±0.01 vs. 3.11±0.04, P <0.05). Laminin expression in serum from the cerebral ischemia group was higher than that in the sham-operated group. Compared with the model group, the level of laminin in the serum of the acupuncture group was significantly lower at each time point, especially at the 3-day, and 7-day time points (589.12±3.57 vs. 793.05±5.28, and 600.53±3.05 vs. 899.06±5.74, P <0.05). The level of integrin β1 in the serum from the acupuncture group was lower than that in the model group particularly at the 3-day and 7-day time points (208.66±0.95 vs. 280.83±1.77, and 212.36±0.95 vs. 316.77±2.42, P <0.05). Additionally, the model group and the acupuncture group showed dual peaks of integrin β1 and laminin expression at 3-day and 7-day.</p><p><b>CONCLUSIONS</b>Acupuncture at GV 20 and ST 36 in rats alleviated CIRI and was associated with upregulated expression of miRNA 124 and with downregulated expression of integrin β1 and laminin in peripheral serum. These changes may represent one of the mechanisms underlying acupuncture's attenuation of CIRI.</p>