背景:金脉通(JMT)是由12种生药组成的中药处方。它已经用于治疗糖尿病性神经性疼痛(DNP)超过30年。
目的:小胶质细胞被认为在神经性疼痛中起重要作用。本研究旨在评估JMT对DNP的保护作用,并探讨小胶质细胞和JAK2/STAT3信号通路的主要参与机制。
方法:使用液相色谱串联质谱法分析JMT的化学成分。使用11至12周龄的雄性Zucker糖尿病脂肪(ZDF)大鼠(fa/fa)构建糖尿病模型。模型大鼠分为5组,分别给予JMT3个剂量(11.6、23.2、46.4g/kg,分别,按原料药材料计算),JAK抑制剂AG490(阳性药物,10μg/天),和安慰剂(去离子水),分别,八周(n=6)。同时,Zucker瘦对照(fa/+)给予安慰剂(n=6)。每周测试体重并每2周监测血糖。使用机械退缩阈值(MWT)和热退缩潜伏期(TWL)测试评估机械异常性疼痛和热痛觉过敏。治疗后,小胶质细胞激活标记Iba-1,CD11B,CD68,神经炎症介质,不同组间比较JAK2/STAT3信号通路的介导因子。通过实时定量PCR和WesternBlot检测靶基因的mRNA和蛋白水平,分别。
结果:我们发现JMT显著抑制脊髓小胶质细胞的过度激活,抑制DNP模型大鼠的神经炎症,从而改善神经功能障碍和损伤。此外,JMT的这些作用可归因于JAK2/STAT3信号通路的抑制.
结论:我们的研究结果表明,JMT通过抑制JAK2/STAT3信号通路调节小胶质细胞活化,从而有效改善DNPJMT。本研究为进一步研究DNP的治疗策略奠定了基础。
BACKGROUND: Jinmaitong (JMT) is a prescription of Traditional Chinese Medicine that is composed of 12 crude drugs. It has been used in the treatment of diabetic neuropathic pain (DNP) for more than 30 years.
OBJECTIVE: Microglia are thought to play an important role in neuropathic pain. This study aimed to evaluate the protective effect of JMT against DNP and to investigate the underlying mechanisms in which the microglia and JAK2/STAT3 signaling pathway were mainly involved.
METHODS: The chemical composition of JMT was analyzed using liquid chromatography tandem mass spectrometry. The diabetes model was constructed using 11 to 12-week-old male Zucker diabetic fatty (ZDF) rat (fa/fa). The model rats were divided into 5 groups and were given JMT at three dosages (11.6, 23.2, and 46.4 g/kg, respectively, calculated as the crude drug materials), JAK inhibitor AG490 (positive drug, 10 μg/day), and placebo (deionized water), respectively, for eight weeks (n = 6). Meanwhile, Zucker lean controls (fa/+) were given a placebo (n = 6). Body weight was tested weekly and blood glucose was monitored every 2 weeks. The mechanical allodynia and heat hyperalgesia were assessed using mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) tests. After treatment, the microglia activation marker Iba-1, CD11B, CD68, neuroinflammatory mediators, and mediators of the JAK2/STAT3 signaling pathway were compared between different groups. The mRNA and protein levels of target genes were assessed by quantitative real-time PCR and Western Blot, respectively.
RESULTS: We found that JMT significantly inhibited the overactivation of microglia in spinal cords, and suppressed neuroinflammation of DNP model rats, thereby ameliorating neurological dysfunction and injuries. Furthermore, these effects of JMT could be attributed to the inhibition of the JAK2/STAT3 signaling pathway.
CONCLUSIONS: Our findings suggested that JMT effectively ameliorated DNP by modulating microglia activation via inhibition of the JAK2/STAT3 signaling pathway. The present study provided a basis for further research on the therapeutic strategies of DNP.