目的: 观察电针对缺血性脑卒中大鼠miR-142-5p及ADAMTS1/磷脂酰肌醇-3激酶（PI3K）/丝氨酸-苏氨酸蛋白激酶（AKT）通路的影响，探讨电针对缺血性脑卒中后血管新生的调控机制。方法: 本研究分为两部分实验。第一部分实验：将SD大鼠随机分为假手术组、模型组、电针组，每组20只。采用改良的Longa法制备大脑中动脉栓塞大鼠模型。电针组于“水沟”进行电针干预（4 Hz/20 Hz），30 min/次，造模成功后即时干预1次，以后每日1次，共4次。采用亨氏评分法评价造模后大鼠神经功能缺损程度；TTC染色法检测大鼠脑梗死体积；HE染色法观察大鼠大脑皮层的病理改变；免疫组织化学法测定脑微血管密度（MVD）；实时荧光定量PCR法和Western blot法检测大鼠大脑皮层缺血半暗带的miR-142-5p及ADAMTS1/PI3K/AKT通路中ADAMTS1、血管内皮生长因子（VEGF）、PI3K、AKT、内皮型一氧化氮合酶（eNOS）mRNA和蛋白表达水平。第二部分实验：将同批次SD大鼠随机分为电针+对照组、电针+miR-142-5p Antagomir（拮抗剂）组，每组8只。电针+对照组给予右侧侧脑室注射0.9%氯化钠溶液，电针+拮抗剂组大鼠造模前30 min给予右侧侧脑室注射miR-142-5p拮抗剂，大鼠注射后进行造模，电针+对照组、电针+拮抗剂组均给予上述同样的电针治疗。实时荧光定量PCR法和Western blot法检测两组大鼠大脑皮层miR-142-5p、ADAMTS1 mRNA及蛋白表达。结果: 与假手术组比较，模型组大鼠神经缺损体征评分升高（P<0.01），脑梗死体积增加（P<0.01），脑组织水肿及神经元坏死程度增加，MVD升高（P<0.01），miR-142-5p表达水平及VEGF、AKT、eNOS mRNA表达水平均降低（P<0.01，P<0.05），VEGF、磷酸化（p）-AKT、eNOS蛋白表达水平均降低（P<0.01），ADAMTS1、PI3K mRNA及ADAMTS1、p-PI3K蛋白表达水平均升高（P<0.01，P<0.05）。与模型组比较，电针组大鼠神经缺损体征评分降低（P<0.01），脑梗死体积缩小（P<0.01），脑组织水肿及神经元坏死程度减轻，MVD升高（P<0.01），miR-142-5p及VEGF、PI3K、AKT、eNOS mRNA的表达水平升高（P<0.01），VEGF、p-PI3K、p-AKT、eNOS蛋白表达水平均升高（P<0.01，P<0.05），ADAMTS1 mRNA及蛋白表达水平降低（P<0.05，P<0.01）。注射miR-142-5p拮抗剂后，与电针+对照组比较，电针+拮抗剂组miR-142-5p表达水平降低（P<0.05），ADAMTS1 mRNA及蛋白表达水平升高（P<0.01，P<0.05）。结论: 电针“水沟”可改善缺血性脑卒中大鼠的神经功能损伤，减少脑梗死体积，促进血管新生，其机制可能是通过促进miR-142-5p表达抑制其靶基因ADAMTS1的表达，上调VEGF表达，从而进一步激活PI3K/AKT通路，促进eNOS释放，促进缺血性脑卒中大鼠脑血管新生。.

2024-07-01·3 Biotech

miR-338-5p regulated the NF-κB/MAPK pathway to alleviate inflammation and oxidative stress by targeting IL-6 in rats with atrial fibrillation

Article

作者: Zhang, Zhen ; Yao, Yali ; Zhang, Yujie ; Wei, Jia

The aim of this study was to investigate the functional effect of miR-338-5p targeting IL-6 on NF-κB/MAPK pathway-mediated inflammation and oxidative stress in atrial fibrillation (AF) rats. AF model rats were generated by tail vein injection of 0.1 mL Ach-CaCl₂ mixture. The overexpression and suppression of miR-338-5p were established by injecting a miR-338-5p-agomir and a miR-338-5p-antagomir, respectively, into AF rats. Cardiac morphological changes were detected by H&E and Masson staining. The levels of ROS, SOD, T-AOC, IL-6, IL-1β, and TNF-α were detected via ELISA. Dual luciferase assays, qRT‒PCR, and western blotting were used to verify that miR-338-5p targets IL-6. The expression of NF-κB/MAPK pathway proteins was detected by western blot. Overexpression of miR-338-5p ameliorated heart damage in AF rats. Increased miR-338-5p reduced the levels of CK, CK-MB, and cTnT to alleviate myocardial injury. Furthermore, overexpression of miR-338-5p relieved inflammation and oxidative stress by downregulating SOD and T-AOC and upregulating IL-6, IL-1β, TNF-α, and ROS. Further research revealed that upregulation of miR-338-5p reduced the protein levels of p-p38, p-p65 and p-ERK1/2. The opposite results were obtained following miR-338-5p-antagomir treatment. Taken together, these findings indicate that the upregulation of miR-338-5p alleviated inflammation and oxidative stress by targeting IL-6 to inhibit the NF-κB/MAPK pathway, thus providing a new therapeutic target for AF.

2024-03-31·Aging

Antagomir of miR-31-5p modulates macrophage polarization via the AMPK/SIRT1/NLRP3 signaling pathway to protect against DSS-induced colitis in mice

Article

作者: Gao, Fei ; Liu, Xingxing ; Fan, Heng ; Yang, Jia ; Yuan, Yuyi ; Wei, Chunzhu ; Zhu, Feng ; Liu, Yujin ; Zhang, Lijuan ; Deng, Shuangjiao ; Chen, Qianyun ; Shou, Zhexing

Macrophage-driven immune dysfunction of the intestinal mucosa is involved in the pathophysiology of ulcerative colitis (UC). Emerging evidence indicates that there is an elevation in miR-31-5p levels in UC, which is accompanied by a downregulation of adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) expression. Nevertheless, the precise influence of miR-31-5p on macrophage polarization and the integrity of the intestinal epithelial barrier in UC remains to be fully elucidated. This study explored the role of miR-31-5p and AMPK in UC through a bioinformatics investigation. It investigated the potential of miR-31-5p antagomir to shift macrophages from pro-inflammatory M1 phenotype to anti-inflammatory M2 phenotype and enhance the intestinal mucosal barrier in DSS-induced UC mice. Additionally, RAW264.7 cells stimulated with LPS were employed to confirm the reversal of miR-31-5p antagomir's therapeutic effect under AMPK inhibition. The findings demonstrated that miR-31-5p antagomir penetrated colonic tissues and ameliorated DSS-induced experimental colitis. Transformation of spleen and mesenteric lymph node macrophages from M1 to M2 type was seen in the DSS+miR-31-5p antagomir group. AMPK/Sirt1 expression increased while NLRP3 expression decreased. Expression of M2-related genes and proteins was enhanced and that of the M1 phenotype suppressed. Tight junction proteins, ZO-1 and occludin, were increased. The therapeutic effects of miR-31-5p antagomir transfection into RAW264.7 cells were repressed when AMPK expression was inhibited. Therefore, our results suggest that suppression of miR-31-5p expression transformed macrophages from M1 to M2, ameliorated inflammation and repaired the intestinal epithelium to alleviate DSS-induced colitis. AMPK/Sirt1/NLRP3 was involved.

100 项与 EBV-miR-BART1-5p-antagomiRs 相关的药物交易

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