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BACKGROUND:Studies have shown that insulin-like growth factor 1/platelet-derived growth factor has an inhibitory effect on fibroblast apoptosis.miR-141-3p in bone marrow stromal cells increases with age and has a relationship with the activation of inflammatory signaling pathways,suggesting that it may be a therapeutic target for lumbar disc herniation. OBJECTIVE:To explore the effects of miR-141-3p on dorsal root ganglion inflammation and lower limb pain in rats with lumbar disc herniation by regulating insulin-like growth factor 1/platelet-derived growth factor. METHODS:Fifty male Sprague-Dawley rats,SPF level,were randomly divided into normal group,model group,miR-NC group,miR-141-3p inhibitor group and miR-141-3p mimics group,with 10 rats in each group.Except for the normal group,animal models of lumbar disc herniation were established in rats by autologous nucleus pulposus transplantation.After successful modeling,rats in the miR-NC,miR-141-3p inhibitor and miR-141-3p mimics groups were injected intrathecally with 10 μL of 20 μmol/L miR-NC,miR-141-3p inhibitor,miR-141-3p mimics,once a day for 28 days,respectively,while those in the normal and model groups were injected with the same volume of saline at the same location at the same time.Paw withdrawal thermal latency threshold was used to evaluate lower limb pain in rats.The mRNA expression of miR-141-3p in dorsal root ganglion tissue was detected by real-time fluorescence quantitative PCR,the levels of inflammatory factors in dorsal root ganglion tissue were detected by ELISA,and the expression of insulin-like growth factor 1/platelet-derived growth factor in dorsal root ganglion tissue was detected by western blot.The correlation between miR-141-3p and insulin-like growth factor 1/platelet-derived growth factor was analyzed. RESULTS AND CONCLUSION:There were no significant differences in all indexes between the miR-NC group and the model group.Paw withdrawal thermal latency threshold was significantly lower in the model group than in the normal group(P＜0.05),significantly lower in the miR-141-3p inhibitor group than the miR-NC group(P＜0.05),and significantly higher in the miR-141-3p mimics group than in the miR-141-3p inhibitor group(P＜0.05).The mRNA expression of miR-141-3p in dorsal root ganglion tissue was significantly lower in the model group than in the normal group(P＜0.05),significantly lower in the miR-141-3p inhibitor group than in the miR-NC group(P＜0.05),and significantly higher in the miR-141-3p mimics group than in the miR-141-3p inhibitor group(P＜0.05).The levels of tumor necrosis factor α,interleukin 1β,and interleukin 1 in dorsal root ganglion tissue were significantly higher in the model group than in the normal group(P＜0.05),significantly higher in the miR-141-3p inhibitor group than in the miR-NC group(P＜0.05),and significantly lower in the miR-141-3p mimics group than in the miR-141-3p inhibitor group(P＜0.05).The protein expressions of insulin-like growth factor 1 and platelet-derived growth factor in dorsal root ganglion tissue were significantly lower in the model group than in the normal group(P＜0.05),significantly lower in the miR-141-3p inhibitor group than in the miR-NC group(P＜0.05),and significantly higher in the miR-141-3p mimics group than in the miR-141-3p inhibitor group(P＜0.05).The expressions of insulin-like growth factor 1 and platelet-derived growth factor showed a positive correlation with miR-141-3p(r=0.904,P＜0.001;r=0.879,P＜0.001).To conclude,miR-141-3p can significantly improve lower limb pain and inhibit inflammation in dorsal root ganglia in rats with lumbar disc herniation,and its mechanism may be related to the promotion of insulin-like growth factor 1/platelet-derived growth factor expression.
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Objective To investigate the relationship between the expression of miR-141-3p,miR-149-3p and proliferative genes,clinicopathological features and prognosis in endometrial carcinoma tissues.Methods From February 2017 to February 2019,98 patients with endometrial cancer were selected as the study objects.Real-time fluorescence quantitative PCR was used to detect the relative mRNA expression levels of miR-141-3p,miR-149-3p and proliferating genes[proliferating cell nuclear antigen(PCNA),cyclin D1,cyclin dependent kinase 4(CDK4)]in cancer tissues of patients with endometrial cancer.The correlation between the expres-sions of miR-141-3p,miR-149-3p and PCNA,cyclin D1 and CDK4 mRNA in cancer tissues of patients with en-dometrial cancer was analyzed by Pearson correlation analysis.The expression of miR-141-3p and miR-149-3p in cancer tissues of endometrial carcinoma patients with different clinicopathological characteristics was com-pared.Kaplan-Meier curve was used to analyze the influence of miR-141-3p and miR-149-3p expression on prognosis of patients with endometrial carcinoma.Univariate and multivariate Cox regression analysis of prog-nostic factors in patients with endometrial cancer.Results The relative expression levels of miR-141-3p,miR-149-3p,PCNA mRNA,cyclinD1 mRNA and CDK4 mRNA in cancer tissues of patients with endometrial carci-noma were higher than those in adjacent tissues,with statistical significance(P＜0.05).The expressions of miR-141-3p and miR-149-3p were positively correlated with the expressions of PCNA mRNA,cyclinD1 mR-NA and CDK4 mRNA in cancer tissues of patients with endometrial carcinoma(all P＜0.05).The relative ex-pression levels of miR-141-3p and miR-149-3p in cancer tissues of endometrial cancer patients with stage Ⅲand lymph node metastasis according to International Federation of Gynecology and Obstetrics(FIGO)were higher than those of endometrial cancer patients with stage Ⅰ-Ⅱ and no lymph node metastasis,respective-ly,and the difference was statistically significant(P＜0.05).The 3-year overall survival rate of miR-141-3p high expression group was significantly lower than that of miR-141-3p low expression group,and the differ-ence was statistically significant(Log-rank x2=7.043,P=0.008).The 3-year overall survival rate of patients with high miR-149-3p expression group was significantly lower than that of patients with low miR-149-3p ex-pression group,with statistical significance(Log-rank x2=7.094,P=0.007).FIGO stage Ⅲ,lymph node me-tastasis,miR-141-3p elevation and miR-149-3p elevation were independent risk factors for prognosis of pa-tients with endometrial cancer(P＜0.05).Conclusion Detecting the expression of miR-141-3p and miR-149-3p in cancer tissues of patients with endometrial carcinoma is helpful to evaluate the survival and prognosis of patients.
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Objective @#To investigate the effect of miR-141-3p on LPS induced A549 cell injury by targeting high mobility group protein 1 (HMGB1) ．@*Methods @#A549 cells derived from type Ⅱ alveolar epithelial cells were taken as the study object，miR-141-3p mimics，mimics NC，HMGB1 gene overexpression plasmid (pcDNA3. 1-HMGB1) and empty Vector were transfected into A549 cells respectively or co-transfected，then 10 μg / ml LPS was used for 24 h．Cell proliferation activity was detected by cell counting kit-8 ( CCK-8) ．The activity of lactate dehydrogenase ( LDH) in the supernatant of cell culture was detected by colorimetry．The apoptosis level of each group was detec- ted by flow cytometry．The levels of interleukin (IL) -1 β , IL-6 and tumor necrosis factor α (TNF-α) were detected by enzyme-linked immunosorbent assay (Elisa) ．Dual luciferase reporter gene assay verified the targeted regulatory relationship between miR-141-3p and HMGB1 . @*Results @#After treatment with LPS ，the proliferative activity of A549 cells and the expression level of miR-141-3p decreased ( P ＜0. 05 ) ，the apoptosis rate increased ( P < 0. 05) ，the levels of IL-1 β , IL-6，TNF-α and the activity of LDH in supernatant increased (P＜0. 05) ．Overex- pression of miR-141-3p increased the proliferation activity of A549 cells treated with LPS (P ＜0. 05 ) ，and de- creased the apoptosis rate and the levels of IL-1 β , IL-6，TNF-α in cells and LDH activity in supernatant (P < 0. 05) ．However，overexpression of HMGB1 gene could reverse the ameliorative effect of miR-141-3p on LPS-in- duced A549 cell injury．Dual luciferase reporter gene experiment confirmed that HMGB1 was the downstream target gene of miR-141-3p．@*Conclusion @# miR-141-3p can inhibit LPS-induced apoptosis，reduce the expression level of inflammatory factors，and improve the damage of A549 cells，which may be related to the targeted regulation of HMGB1 expression.
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Objective To investigate the effect and mechanism of miR-141-3p on pulmonary fibrosis in rats with acute respiratory distress syndrome(ARDS).Methods Rats were divided into the control group,the model group,the agomir negative control group and the miR-141-3p agomir group according to random number table,with 10 rats in each group.In addition to the control group,the ARDS rat model was established by lipopolysaccharide(LPS)infusion.Rat alveolar typeⅡepithelial cells RLE-6TN cells were divided into the NC group,the LPS group,the miR-NC group,the miR-141-3p mimics group,the miR-141-3p mimics+pcDNA group and the miR-141-3p mimics+NRF2 and Kelch-like ring associated protein 1(Keap1)group.LPS cell model was established in all groups except the NC group.The mRNA expression levels of miR-141-3p and Keap1 in lung tissue and cells were detected by qPCR.Western blot assay was used to analyze lung tissue and cell epithelial cadherin(E-cadherin),neural cadherin(N-cadherin),microtubule associated protein light chain 3B(LC3B),autophagy associated gene Beclin-1,α-smooth muscle actin(α-SMA),type I collagen(Col-Ⅰ),Keap1 and nuclear factors E2 related factor 2(NRF2)and heme oxygenase 1(HO-1).HE staining and Masson staining were used to observe pathological changes of lung tissue and to estimate the area of lung tissue injury and pulmonary fibrosis.Hydroxyproline(Hyp)in lung tissue was detected by the kit.Levels of inflammatory factor interleukin-1β,tumor necrosis factor(TNF-α)and oxidative stress index malondialdehyde(MDA)and superoxide dismutase(SOD)were detected by ELISA.Dual luciferase reporting experiment was used to verify the targeting relationship between miR-141-3p and Keap1.Results The expression of miR-141-3p was down-regulated and the expression of Keap1 was up-regulated in lung tissue and cells(P＜0.05).Overexpression of miR-141-3p can reduce the degree of pathological damage and fibrosis of lung tissue in rats,Hyp content,and up-regulate expression levels of SOD,E-cadherin,LC3B,Beclin-1,NRF2 and HO-1 in lung tissue and cells,and down-regulate the expression levels of IL-1β,TNF-α,MDA,N-cadherin,α-SMA,Col-I and Keap1(P＜0.05).Overexpression of Keap1 was able to reverse the improvement effect of overexpression of miR-141-3p on alveolar epithelial cell damage in ARDS rats(P＜0.05).Double Luciferase reporter gene experiment confirmed that miR-141-3p and Keap1 may have a targeted regulatory relationship.Conclusion Overexpression of miR-141-3p may activate the Keap1-NRF2/ARE signaling pathway,activate autophagy,inhibit inflammatory response,oxidative stress,and EMT progression,and improve pulmonary fibrosis in ARDS rats.
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AIM: To explore the relationship between the changes of serum circFTO and microRNA-141-3p(miR-141-3p)levels and the different disease stages of diabetes retinopathy.METHODS: A total of 198 patients with type 2 diabetes admitted to our hospital from October 2019 to November 2022 were collected as the study subjects, the patients were grouped into non diabetes retinopathy(NDR)group(70 cases), non proliferative diabetes retinopathy(NPDR)group(66 cases)and proliferative diabetes retinopathy(PDR)group(62 cases)according to different stages; meantime, 67 volunteers with normal physical examination results were collected as the control group. The levels of serum circFTO and miR-141-3p were detected by real-time fluorescent quantitative PCR(qRT-PCR); Pearson correlation analysis was used to examine the correlation between the serum circFTO, miR-141-3p and various indicators in patients with diabetes retinopathy; multivariate Logistic regression analysis was applied to explore the influencing factors of diabetes retinopathy.RESULTS: CircFTO, systolic blood pressure(SBP), and diastolic blood pressure(DBP)in PDR group were higher than those in control group, NDR group and NPDR group, while miR-141-3p and high-density lipoprotein cholesterol(HDL-C)were lower than those in control group, NDR group and NPDR group(P<0.05). Fasting blood glucose(FPG)and glycosylated hemoglobin(HbA1c)in NDR group, NPDR group and PDR group were higher than those in the control group(all P<0.05). The course of disease in PDR group was longer than that in NDR group and NPDR group(P<0.05). Serum circFTO in patients with diabetes retinopathy was positively correlated with SBP, DBP, FPG, HbA1c, and miR-141-3p was negatively correlated with SBP, DBP, FPG, HbA1c(all P<0.05). CircFTO was a risk factor for diabetes retinopathy, and miR-141-3p was a protective factor for diabetes retinopathy(P<0.05).CONCLUSION: Serum circFTO is obviously increased and miR-141-3p is obviously decreased in patients with diabetes retinopathy, both of them are closely related to disease stage, and are expected to become important indicators for evaluating disease progress.
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OBJECTIVE：To study the improvement effects of icariside Ⅱ（ICS Ⅱ）on neurological function of focal cerebral ischemia model rats by regulating miR- 141-3p/Notch/nuclear factor erythroid- 2-related factor 2（Nrf2）axis（miR-141-3p/Notch/ Nrf2）. METHODS ：The rats were divided into sham operation group ，model group ，nimodipine group （20 mg/kg）and ICS Ⅱ low-dose，medium-dose and high-dose groups （4，8，16 mg/kg），with 20 rats in each group. Twenty-four hours after establishing focal cerebral ischemia model ，model rats were given re levant medicine or normal saline intragastrically ，twice a day ，for consecutive 3 d. The neurological deficit of rats in each group was scored ；the volume of cerebral infarction was measured by 2，3， 5-triphenyltetrazolium chloride （TTC）staining；water content of cerebral tissue and the permeability of blood-brain barrier were measured；HE staining was performed to observe the pathological change of cerebral tissue of rats ；the expression of miR- 141-3p in cerebral tissue of rats was measured by qRT-PCR ；the protein expression of Notch and Nrf 2 in cerebral tissue of rats were measured by Western blotting assay. RESULTS ：Compared with sham operation group ，the neurological deficit score ，expression of Notch-1 and Nrf 2 in model group were significantly lowered （P＜0.05）；infarction volume ，brain water content ，the permeability of blood-brain barrier and the expression of miR- 141-3p in cerebral tissue were increased significantly （P＜0.05）；the distribution of cortical cells was disordered ，and inflammatory infiltration and necrosis were observed in a large number of nerve cells. Compared with model group ，the neurological deficit score ，the protein expression of Notch- 1 and Nrf 2 in cerebral tissue were significantly increased in ICS Ⅱgroups（P＜0.05）；infarction volume ，brain water content ，the permeability of blood-brain barrier and the expression of miR- 141-3p in cerebral tissue were decreased significantly （P＜0.05）；the arrangement of cortical cells was regular，and the inflammatory infiltration and necrosis of nerve cells were decreased significantly. CONCLUSIONS ：ICS Ⅱ can promote the recovery of neurological function in focal cerebral ischemic model rats ，which may be related to down-regulation of miR-141-3p and activation of Notch/Nrf 2 axis.
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@#Objective: To investigate the regulatory effect of lncRNA MALAT1/miR-141-3p/ZEB1 axis on the invasion, metastasis and epithelial mesenchymal transition (EMT) of gastric cancer (GC) cells. Methods: Thirty-eight pairs of GC tissues (non-necrotic part) and corresponding adjacent tissues (>5 cm away from tumor tissue) removed by general surgery in Wuhan Commercial Hospital from April 2014 to May 2017 were collected. Meanwhile, normal gastric epithelial GES1 cells and GC cell lines (SGC7901, HGC27, BGC823, MKN45 and MKN28) were selected. The expression level of MALAT1 and miR-141-3p in GC tissues and cell lines were detected by qPCR. The effect of MALAT1 knockdown on proliferation, migration and invasion of SGC7901 cells was determined by CCK-8 assay and Transwell assay. WB was performed for measuring the expression level of ZEB1, E-cadherin, N-cadherin and Vimentin. Dual luciferase reporter gene assay was used to validate the relationship among MALAT1, miR-141-3p and ZEB1. CCK-8 assay and Transwell assay were used to detect the effect of MALAT1/miR-141-3p/ZEB1 axis on biological behaviors of SGC7901 cells. Results: MALAT1 was over-expressed in GC tissues and cell lines (P<0.05 or P<0.01). Knockdown of MALAT1 significantly inhibited the proliferation, migration, invasion and EMT of SGC7901 cells (P<0.05 or P<0.01). The results of dual luciferase reporter gene assay showed that MALAT1 directly targeted miR-141-3p, as well as for miR-141-3p and ZEB1. Further experiment indicated that simultaneous over-expression of miR-141-3p and MALAT1 or ZEB1 could restore the biological behaviors of SGC7901 cells, which were inhibited by miR-141-3p. Conclusion: MALAT1 promotes the invasion, metastasis and EMT of GC SGC7901 cells by down-regulating the inhibitory effect of miR-141-3p on ZEB1.
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@#Objective: To explore the effect of miR-141-3p on the proliferation, invasion and apoptosis of ovarian cancer cells via targeting PTEN and regulating PI3K/Akt pathway. Methods: Collecting twenty-eight cases pairs of ovarian cancerovarian cancer patients with tumor tissues and adjacent tissues were collected from patients, who from April 2014 to October 2017 were treated in the Department of Obstetrics and Gynecology. qPCR was applied to detect the expression of miR-141-3p in ovarian cancer tissues and cell lines. The relationship between miR-141-3p and PTEN was verified by dual-luciferase reporter gene assay. After over-expression or knockdown of miR-141 and PTEN genes, the cell viability, invasion and apoptosis of ovarian cancer A2780 cells were examined by CCK-8 assay, Transwell assay and Annexin V-FITC/PI double staining flow cytometry assay, respectively. Furthermore, the effect of miR-1413p on PTEN-PI3K/Akt signaling pathway was measured by WB. Results: miR-141-3p is was highly expressed in ovarian cancer tissues and cell lines (P<0.05 or P<0.01). The dual luciferase reporter gene assay confirmed that miR-141-3p targets PTEN was a target of miR-141-3p and downregulates its expression level was down-regulated (P<0.01). Compared with the control group, after knockdown of miR-141-3p, the proliferation ofA2780 cells was significantly inhibited after knockdown of miR-141-3p (at 48 h, 0.36±0.04 vs 0.82± 0.06, P<0.05), and the invasive ability of A2780 cells was significantly reduced (number of transmembrane cells: 215.32±16.04 vs 45.14±7.88, P<0.01), while the apoptotic rate was significantly increased ([1.85±0.26]% vs [9.29±0.65]%, P<0.01). Over-expression of PTEN significantly inhibited the expression of p-Akt and cell proliferation and invasion in A2780 cells (all P<0.01), inhibited cell proliferation and invasion (all P<0.01) and significantly promoted apoptosis (all P<0.01). However, simultaneous over-expression of miR141-3p or addition of IGF-1 wile over-expressing PTEN can offset the above effects. Conclusion: miR-141-3p facilitates the proliferation, invasion and decreases apoptosis of A2780 cells. The mechanism may be related to targeted regulation of PTEN and activation of PI3K/Akt pathway.
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@#Objective: To investigate the relationship between miR-141-3p and transforming growth factorβ2 (TGF-β2), and its effects on the malignant biological behaviors of human prostate cancer cell line C4-2B. Methods:After the transfection of miR-141-3p mimic, the mRNAexpression of miR-141-3p and TGF-β2 in C4-2B cells was detected by qRT-PCR. Bioinformatics method validated the relationship between miR-141-3p and TGF-β2. miR-141-3p mimic alone or with TGF-β2 over-expression vector was transfected into C42B cells, and then Western blotting was used to detect the expression of TGF-β2 protein in C4-2B cells, Hochest33258 staining was used to detect cell apoptosis, and Transwell assay was used to detect the invasion ability of cells in each group. Results:After the transfection of C4-2B cells with miR-141-3p mimic, the level of miR-141-3p increased significantly, and the level of TGF-β2 mRNA decreased significantly (all P<0.01). The activity of luciferase was significantly reduced after the co-transfection with miR-141-3p mimic and wild type report plasmid (P<0.01); However, the activity of luciferase was not obviously changed after co-transfection with miR141-3p mimic and mutant type report plasmid (P>0.05).After co-transfection with miR-141-3p mimic and pc-TGF-β2, the proliferation of C4-2B cells decreased significantly, the number of apoptotic cells increased significantly, and the cell invasion ability decreased significantly (all P<0.01). Conclusion: miR-141-3p inhibits the proliferation and invasion of human prostate cancer C4-2B cells and induces cell apoptosis by targeting TGF-β2.