First Clinical Medical College, Anhui University of Chinese Medicine, Hefei, Anhui, 230012, People's Republic of China.
Department of Rheumatology, First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei, Anhui, 230031, People's Republic of China.
Drug Des Devel Ther. 2024 Jun 20;18:2421-2433. doi: 10.2147/DDDT.S456783. eCollection 2024.
This study probed the mechanism of action of Xinfeng Capsule (XFC) in myocardial injury in rats with adjuvant arthritis (AA) via the growth arrest-specific transcript 5 (GAS5)/microRNA-21 (miR-21)/Toll-like receptor 4 (TLR4) axis.
Rats were injected with Freund's complete adjuvant to establish a rat model of AA. Then, some modeled rats were given normal saline or drugs only, and some modeled rats were injected with adeno-associated viruses or necrosulfonamide (NSA; a pyroptosis inhibitor) before drug administration. Toe swelling and arthritis index (AI) were calculated. Pathological and morphological changes in synovial and myocardial tissues were analyzed with hematoxylin-eosin staining, and pyroptotic vesicles and the ultrastructural changes of myocardial tissues were observed with transmission electron microscopy. The serum levels of interleukin (IL)-1β, IL-18, IL-6, and tumor necrosis factor (TNF)-α were detected, and lactate dehydrogenase (LDH) release was measured in myocardial tissues, accompanied by the examination of GAS5, miR-21, TLR4, nuclear factor-kB (NF-κB) p65, nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), Caspase-1, and Gasdermin D (GSDMD) expression in myocardial tissues.
After AA modeling, rats presented with significantly increased toe swelling and AI scores, synovial and myocardial tissue damage, elevated pyroptotic vesicles, and markedly enhanced serum levels of IL-1β, IL-18, IL-6, and TNF-α, accompanied by significantly diminished GAS5 expression, substantially augmented miR-21, TLR4, NF-κB p65, NLRP3, Caspase-1, and GSDMD expression, greatly increased LDH release in myocardial tissues. XFC treatment significantly declined toe swelling, AI scores, synovial and myocardial tissue damage, and the serum levels of IL-1β, IL-18, IL-6, and TNF-α in AA rats. Additionally, XFC treatment markedly elevated GAS5 expression and substantially lowered LDH release and miR-21, TLR4, NF-κB p65, NLRP3, Caspase-1, and GSDMD expression in myocardial tissues of AA rats. Moreover, the above effects of XFC in AA rats were further promoted by GAS5 overexpression or NSA treatment.
XFC alleviated myocardial injury in AA rats by regulating the GAS5/miR-21/TLR4 axis and inhibiting pyroptosis and pro-inflammatory cytokine secretion.
本研究通过生长停滞特异性转录物 5（GAS5）/微小 RNA-21（miR-21）/Toll 样受体 4（TLR4）轴探讨新风胶囊（XFC）在佐剂性关节炎（AA）大鼠心肌损伤中的作用机制。
用弗氏完全佐剂给大鼠注射建立 AA 大鼠模型。然后，部分造模大鼠给予生理盐水或仅给予药物，部分造模大鼠在给予药物前给予腺相关病毒或坏死磺胺（NSA；一种细胞焦亡抑制剂）。计算趾肿胀和关节炎指数（AI）。苏木精-伊红染色分析滑膜和心肌组织的病理和形态变化，透射电镜观察细胞焦亡小泡和心肌组织的超微结构变化。检测血清白细胞介素（IL）-1β、IL-18、IL-6 和肿瘤坏死因子（TNF）-α水平，测量心肌组织中乳酸脱氢酶（LDH）释放量，同时检测心肌组织中 GAS5、miR-21、TLR4、核因子-κB（NF-κB）p65、核苷酸结合寡聚化结构域样受体蛋白 3（NLRP3）、Caspase-1 和 Gasdermin D（GSDMD）表达。
AA 造模后，大鼠的趾肿胀和 AI 评分明显升高，滑膜和心肌组织损伤，细胞焦亡小泡明显增多，血清中 IL-1β、IL-18、IL-6 和 TNF-α水平明显升高，GAS5 表达明显降低，miR-21、TLR4、NF-κB p65、NLRP3、Caspase-1 和 GSDMD 表达明显升高，心肌组织中 LDH 释放明显增加。XFC 治疗可明显降低 AA 大鼠的趾肿胀、AI 评分、滑膜和心肌组织损伤以及血清中 IL-1β、IL-18、IL-6 和 TNF-α水平。此外，XFC 治疗还可明显升高 AA 大鼠心肌组织中 GAS5 表达，降低 LDH 释放和 miR-21、TLR4、NF-κB p65、NLRP3、Caspase-1 和 GSDMD 的表达。而且，用 GAS5 过表达或 NSA 处理可进一步促进 XFC 在 AA 大鼠中的上述作用。
XFC 通过调节 GAS5/miR-21/TLR4 轴抑制细胞焦亡和促炎细胞因子的分泌，缓解 AA 大鼠的心肌损伤。
