Search

BACKGROUND: First-trimester human decidual mesenchymal stem cells and tanshinone IIA have anti-adhesion effect, and both have a certain effect on intrauterine adhesion, but the specific mechanism needs to be further studied. OBJECTIVE: To explore the efficacy and mechanism of first-trimester human decidual mesenchymal stem cells combined with tanshinone IIA in the treatment of intrauterine adhesions. METHODS: First-trimester human decidual mesenchymal stem cells were isolated and cultured by collagenase I digestion. A total of 45 female SD rats aged 4-6 weeks were randomly divided into 5 groups with 9 rats in each group: control group, model group, tanshinone group, stem cell group and combined treatment group. Sham operation was performed in the control group, and no intervention was performed in the model group after modeling. In the stem cell group, 0.2 mL human decidual mesenchymal stem cell suspension (1×109 L-1) was injected parauterine 7 days after modeling, once every other 3 days, for a total of 3 times. In the tanshinone group, tanshinone IIA solution at 11 mg/(kg·d) was given via intragastulation for 1 week. The combined treatment group received the combination of the above two treatment methods. In the control group and model group, samples were obtained at 5, 10, and 15 days after operation and sham operation. In each treatment group, samples were taken at 5, 10, and 15 days after the last treatment. Hematoxylin-eosin staining and Masson staining were performed to observe the healing degree of the damaged uterus. Serum vascular endothelial growth factor and E2 levels were detected by ELISA. q-PCR was used to detect the expression of miR-29a mRNA in uterine tissue. The protein expression levels of transforming growth factor β1, Smad3 and matrix metalloproteinase 9 were detected by western blot assay. RESULTS AND CONCLUSION: (1) Compared with the control group, the endometrial fibrosis area ratio in the model group increased significantly (P < 0.05), while the endometrial thickness and the number of glands decreased significantly (P < 0.05). Compared with the model group, the degree of endometrial fibrosis in the combined treatment group was significantly lower (P < 0.05) and the thickness of the intima and the number of glands increased significantly (P < 0.05). The endometrial fibrosis area ratio of rats in the stem cell group and the tanshinone group also decreased significantly (P < 0.05). (2) Compared with the model group, the serum estrogen E2 level of the tanshinone, stem cell and the combined treatment groups was significantly increased (P < 0.05); vascular endothelial growth factor level was significantly decreased (P < 0.05); the expression of miR-29a mRNA was significantly increased (P < 0.05); the expression of transforming growth factor β1 and Smad3 protein in the combined treatment group was significantly decreased (P < 0.05), and the expression of matrix metalloproteinase 9 protein in the tanshinone group was significantly increased (P < 0.05). (3) Results suggest that the combined treatment of first-trimester human decidual mesenchymal stem cells and tanshinone IIA can effectively repair the endometrium in rats, and the mechanism may be through the regulation of miR-29a to reduce the levels of transforming growth factor β1 and Smad3 protein, and then improve the fibrosis. [ABSTRACT FROM AUTHOR]

BACKGROUND: For type 2 diabetes mellitus accompanied by myocardial interstitial fibrosis, although miR-29a closely regulates the transforming growth factor (TGF)-β1/Smads pathway, little is reported about this signaling pathway regulating myocardial interstitial fibrosis in type 2 diabetes. Existing studies have shown that exercise can improve myocardial fibrosis in type 2 diabetes. OBJECTIVE: To investigate the effect of different kinds of exercises on myocardial fibrosis in spontaneous type 2 diabetic mice (KK-Ay mice) and the molecular regulation of miR-29a/TGF-β1 pathway in this process. METHODS: Twenty-four KK-Ay mice, 10 weeks of age, were adaptively fed for 1 week, and randomly divided into KK-Ay control group (KC; 8 mice), KK-Ay swimming group (KY; 8 mice) and KK-Ay treadmill group (KP; 8 mice). Another eight C57BL/6J mice, 10 weeks of age, were selected as normal control group (ZC). KK-Ay mice were given a high-fat diet, and C57BL/6J mice were fed with common feed. The mice in the KY and KP groups were trained for 8 weeks by swimming and running exercise, respectively. Afterwards, the wall of the left ventricle of the mouse was taken, and the change of myocardial fibrosis was detected by Masson staining. The mRNA expression of relevant factors in the myocardium was detected by RT-PCR; the protein level and expression of the relevant factors in the myocardium were detected by ELISA and western blot, respectively. The study protocol was approved by the Animal Ethics Committee of Physical Education School of Yangzhou University with an approval No. YZU-TYXY-31. RESULTS AND CONCLUSION: Compared with the ZC group, the KC group had a significant degree of myocardial interstitial fibrosis; miR-29a mRNA expression and protein level in the myocardium were significantly reduced and TGF-β1, Smad3, Smad4, type I collagen mRNA expression and p-Smad2, p-Smad3, p-Smad4 protein expression were significantly up-regulated (P < 0.05 or P < 0.01), and TGF-β1 and type I collagen protein levels were significantly increased (P < 0.05 or P < 0.01). Compared with the KC group, no significant change in myocardial fibrosis was observed in the KY group; the expression of TGF-β1 and Smad4 mRNAs in the myocardium was significantly down-regulated and the TGF-β1 protein level was significantly reduced (P < 0.05), and other related indicators were not significantly changed. The degree of myocardial fibrosis in the KP group was significantly improved; miR-29a mRNA expression and protein level in the myocardium increased (P < 0.05), TGF-β1, Smad2, Smad3 and COL1 mRNA expression and TGF-β1, COL1 protein levels decreased significantly (P < 0.05 or P < 0.01), and p-Smad2, p-Smad3 and p-Smad4 protein expression was significantly down-regulated (P < 0.05). Compared with the KY group, the myocardial fibrosis of the KP group was significantly improved; miR-29a mRNA expression and protein level in the myocardium were significantly increased (P < 0.05), TGF-β1, COL1 mRNA expression and protein level were significantly reduced (P < 0.05), and p-Smad2, p-Smad3 and p-Smad4 protein expression was significantly downregulated (P < 0.05 or P < 0.01). To conclude, type 2 diabetic mice have severe myocardial fibrosis. Treadmill exercise inhibits the TGF-β1/Smads pathway by upregulating miR-29a in the myocardium of T2DM mice to improve myocardial fibrosis, but swimming exercise has no significant effect. [ABSTRACT FROM AUTHOR]

Objective: To investigate the protective effect of miR-29a on synovial injury in rats with knee osteoarthritis (KOA). A KOA rat model was established by anterior cruciate ligament transection (ACLT). Rats were injected with microRNA- negative control and miR-29a. miR-29a expression in synovial tissue and synovial cells of KOA was detected by real-time quantitative polymerase chain reaction (RT-qPCR). Toll-like receptor 4/ myeloid differentiation protein 88/ nuclear factor κB (TLR4/MyD88/NF-κB) signaling pathway related protein expression was detected by RT-qPCR and Western blot assay. The expression levels of inflammatory cytokines in KOA synovial tissue and synovial cells were detected. Results: miR-29a expression was down-regulated in KOA synovial tissue and synovial cells. Up-regulation of Mir-29a inhibits the inflammatory response of synoviocytes in KOA rats, and inactivates the TLR4/Myd88/NF-κB signaling pathway in KOA rats. Conclusion: The up-regulation of miR-29a inhibited the inflammatory response of synovial cells in KOA rats through the inactivation of TLR4/MyD88/NF-κB signaling pathway, thereby protecting the synovial injury. [ABSTRACT FROM AUTHOR]

Copyright of Practical Pharmacy & Clinical Remedies is the property of Editorial Department of Practical Pharmacy & Clinical Remedies and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

摘 要： 【目的】 探讨微小ＲＮＡ２９ａ（ｍｉＲ－２９ａ）在小鼠子宫内膜基质细胞（ＥＳＣ）蜕膜化中的作用。【方法】 收集孕第１～９天（Ｄ１ ～ Ｄ９）的小鼠子宫，通过实时定量ＰＣＲ（ｑＲＴ－ＰＣＲ）的方法检测小鼠子宫组织中ｍｉＲ－２９ａ的表达；构建去除卵巢及其甾体激素的处理模型，观察甾体激素对ｍｉＲ－２９ａ表达的影响。采用雌二醇（Ｅ２）和孕酮（Ｐ４）对分离培养的小鼠子宫内膜基质细胞进行蜕膜化处理，并通过瞬时转染的方法下调ｍｉＲ－２９ａ的表达，分别收集蜕膜化处理不同时间及干扰前后的细胞，采用ｑＲＴ－ＰＣＲ检测催乳素相关蛋白（ｄＰＲＰ）ｍＲＮＡ的表达，Ｗｅｓｔｅｒｎ ｂｌｏｔ法检测细胞周期蛋白Ｄ３（ｃｙｃｌｉｎ Ｄ３）、孕酮受体（ＰＲ）蛋白的表达。 【结果】 在小鼠妊娠第１～５ 天，子宫组织中的ｍｉＲ－２９ａ 表达量较低，随着蜕膜化的进程（妊娠第６～９天），ｍｉＲ－２９ａ的表达量均显著性增加（与妊娠Ｄ１相比）；在去除卵巢及其甾体激素的处理模型中，一旦给予雌孕激素刺激，ｍｉＲ－２９ａ的表达量显著增加；在小鼠子宫内膜基质细胞体外诱导蜕膜化过程中，ｑＲＴ－ＰＣＲ结果显示ｄＰＲＰ ｍＲＮＡ的表达以及ｍｉＲ－２９ａ的表达随着蜕膜化时间的延长逐渐升高（与２４ ｈ相比），Ｗｅｓｔｅｒｎ ｂｌｏｔ法显示蜕膜化标志分子ｃｙｃｌｉｎ Ｄ３、ＰＲ的表达也随着蜕膜化时间的延长逐渐升高；转染下调ｍｉＲ－２９ａ后，蜕膜化标志分子及ｍｉＲ－２９ａ的表达也相应下降。【结论】 妊娠早期ｍｉＲ－２９ａ表达于母胎界面，且受子宫内膜蜕膜化的影响，说明ｍｉＲ－２９ａ具有促进子宫内膜蜕膜化的作用。

Objective: CRISPR-C'as9 technology was used to edit miR-29a gene in GT1-7 cells and construct miR-29a knockotit GT1-7 cell model. Methods: To induce mutation on miR-29a loci. Cas9 steadily expressed GT1-7 cells were constnicted and transfected by sgRNA plasmids. Tlien a plasmid co-expressing EGFP and sgRNA was constructed and transfected into Cas9 expressing cells, flow cytometry was applied to enrich EGFP positive cells and test positive Monoclonal cell. Finally RT-PCR was used to detect the expression levels of miR-29a in enriched cells and Monoclonal cell. Results: Mutation could be efficiently induced on miR-29a loci by CRISPR-Cas9 system detected by T7E1 assays. Compared with control group, the expression of miR-29a in enriched positive cells totally decreased by 50 percent (P<0.05) using RT-PC'R. Furthermore, one homogeneous miR-29a gene knockout cell clone was obtained by flow screening. The miR-29a expression of this cell clone decreased by 75 percent (P<0.05)compared with control group. Conclusions: An effective gene edit- ing method in GT1-7 cells was established, which constructed a miR-29a steadily knockout cell model. [ABSTRACT FROM AUTHOR]

Objective To investigate the expression levels of microRNA(miR)-125b, miR-29a and miR-155-5p in cerebrospinal fluid (CSF) and plasma from the patients with tuberculous meningitis and their clinic significance. Methods Cerebrospinal fluid and plasma samples were collected from 20 patients with tuberculous meningitis (tuberculous meningitis group) and 20 patients suffered from primary headache (control group). The total RNAs were extracted.The levels of miR- 125b, miR-29a and miR-155-5p were determined by real-time quantitative PCR (q-PCR). Results Levels of miR-29a and miR-125b in both CSF and plasma of patients in tuberculous meningitis group were significantly higher than those in patients from control group with statistical significance (P<0.01). Mean time, the level of miR-155-5p in plasma but not in CSF of patients in tuberculous meningitis group was higher than those in control group with statistical significance (P< 0.01). Conclusion Expression of miR-125b, miR-29a and miR-155-5p may participate in regulating the occurrence and development of tuberculous meningitis. And miR-125b and miR-29a may be used as potential biomarkers for diagnosing tuberculous meningitis. [ABSTRACT FROM AUTHOR]

目的评价血清miR-103a、miR-30b、miR-29a相对表达量在帕金森病中的诊断效能。方法采用RTPCR法检测90例帕金森病患者(观察组)及24例健康人(对照组)外周血清中miR-103a、miR-30b、miR-29a。采用受试者工作曲线法观察血清miR-103a、miR-30b、miR-29a用于诊断帕金森病的效能。结果观察组外周血清miR-103a、miR-30b、miR-29a相对表达量分别为1.12±0.10、1.57±0.37、1.23±0.09,对照组分别为0.43±0.03、0.97±0.07、0.51±0.04,观察组外周血清miR-103a、miR-30b、miR-29a相对表达量均高于对照组,P均<0.05。外周血清miR-103a诊断帕金森病的AUC为0.816,选择0.715为诊断阈值时约登指数最大,此时外周血清miR-103a诊断帕金森病的灵敏度为71.3%,特异度为83.9%。外周血清miR-30b诊断帕金森病的AUC为0.789,选择1.120为诊断阈值时约登指数最大,此时外周血清miR-30b诊断帕金森病的灵敏度为70.5%,特异度为84.6%。外周血清miR-29a诊断帕金森病的AUC为0.756,选择1.001为诊断阈值时约登指数最大,此时外周血清miR-29a诊断帕金森病的灵敏度为72.1%,特异度为80.8%。结论帕金森病患者外周血清miR-103a、miR-30b、miR-29a相对表达量升高。外周血清miR-103a、miR-30b、miR-29a相对表达量在帕金森疾病中的诊断效能好。 [ABSTRACT FROM AUTHOR]

The article presents a study which aims to determine the effect of microRNA expression in the neuronal dysfunction caused by rabies virus (RABV) infection using the bioinformatics and gene analysis method.