背景:糖尿病周围神经病变(DPN)是糖尿病的常见并发症,主要表现为感觉异常。唐祖颗粒(TZG)是由著名的中药汤剂制成,经过长期的临时实践优化。TZG在改善麻木方面有很好的疗效,DPN患者下肢疼痛和瘙痒。然而,其对DPN影响的总体调节机制尚不清楚.
目的:本研究旨在探讨TZG治疗DPN的潜在机制。
方法:Sprague-Dawley(SD)大鼠采用链脲佐菌素(STZ)注射和高脂饮食(HFD)喂养建立DPN体内模型。此外,体外高糖诱导坐骨神经胶质RSC96细胞。干预组SD大鼠接受TZG治疗12周。治疗12周后,采用智能热板仪和神经电生理检测仪评价坐骨神经功能。用苏木精-伊红染色和透射电镜观察坐骨神经细胞的形态学变化。IL-1β,IL-18炎性细胞因子,蛋白质印迹法观察到细胞凋亡和P2X7R/NLRP3信号通路,免疫荧光染色和ELISA。
结果:TZG改善DPN大鼠的神经传导速度和坐骨神经病变的合理结构改变。它还抑制由高糖诱导的RSC96炎症反应和细胞死亡。这可能与TZG抑制P2X7R有关,降低NLRP3炎性体的激活,下调焦亡蛋白的水平,如caspase-1,calvedcaspase-1,gasderminD(GSDMD),和GSDMD-N,并抑制白细胞介素(IL)-18和IL-1β炎性细胞因子的释放。
结论:TZG通过P2X7R/NLRP3信号通路抑制细胞凋亡,缓解神经炎症,对DPN的治疗具有保护作用。
BACKGROUND: Diabetic peripheral neuropathy (DPN) is a common complication of diabetes mellitus, mainly manifested as paresthesia. Tangzu granule (TZG) is derived from famous traditional Chinese medicine decoctions and optimized by long-term temporary practice. TZG has good efficacy in improving numbness, pain and pruritus of the lower extremities of DPN patients. However, the overall regulatory mechanisms underlying its effects on DPN remain unclear.
OBJECTIVE: This study aims to explore the potential mechanism of TZG for treating DPN.
METHODS: Sprague-Dawley (SD) rats were used to establish an in vivo model of DPN with streptozotocin (STZ) injection and high-fat diet (HFD) feeding. Additionally, sciatic glial RSC96 cells were induced with high glucose in vitro. SD rats in intervention group received TZG treatment for 12 weeks. After 12 weeks of treatment, sciatic nerve function was evaluated by intelligent hot plate meter and neuro electrophysiology detector. The morphological changes of sciatic nerve cells were observed by hematoxylin-eosin staining and transmission electron microscope. IL-1β, IL-18 inflammatory cytokines, pyroptosis and P2X7R/NLRP3 signaling pathway were observed by Western blotting, immunofluorescence staining and ELISA.
RESULTS: TZG improved nerve conduction velocity and sciatic neuropathy rational structural changes in DPN rats. It also inhibited RSC96 inflammatory response and cell death that induced by high glucose. This may be related to TZG inhibiting P2X7R, decreasing the activation of NLRP3 inflammasomes, down-regulating the levels of pyroptosis proteins such as caspase-1, cleaved caspase-1, gasdermin D (GSDMD), and GSDMD-N, and inhibiting the release of interleuki (IL)-18 and IL-1β inflammatory cytokines.
CONCLUSIONS: TZG inhibited pyroptosis through P2X7R/NLRP3 signaling pathway, alleviated neuroinflammation, and showed protective effect in the treatment of DPN.