Wu Jiao, Xie Wen, Xie Yucen, Mazhar Maryam, Duan Junguo
Eye School of Chengdu University of TCM, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
Department of Cardiology, Affiliated Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, China.
Front Pharmacol. 2025 May 21;16:1573693. doi: 10.3389/fphar.2025.1573693. eCollection 2025.
Zhilong Huoxue Tongyu Capsule (ZLHXTY) has been used in clinical treatment of vascular diseases caused by hypertension over 20 years. However, the specific mechanisms by ZLHXTY alleviate hypertensive retinopathy (HR) needs to be further explored.
HR mouse model was established by infusing Ang II via subcutaneously implanted osmotic mini-pumps, followed by oral administration of ZLHXTY (0.35, 0.7, 1.4 g/kg/day) 28 days for treatment. To assess the impacts of ZLHXTY on retinal neurodegeneration and vascular injury, multiple experiments such as OCTA, ERG and HE staining were performed. Subsequently, network pharmacological and 4D-label-free proteomics to clarify the potential targets and mechanisms of ZLHXTY alleviated HR. Finally, Western blot, ELISA, IF, and other techniques were utilized to detect the expression of proteins related to inflammation, oxidative stress, and NLRP3 inflammasome activation.
ZLHXTY significantly alleviated retinal dysfunction, increased retinal blood flow, and mitigated pathological changes such as retinal tissues edema in HR mice. Network pharmacology indicated that ZLHXTY might exert anti-inflammatory and anti-oxidative stress effects through targets such as TNF and NF-κB. Proteomic analysis showed that the differential proteins between the ZL group and the Ang II group were mainly enriched in the immune-inflammatory response, and the main mechanism of which might be related to the assembly of NLRP3 inflammasome. Subsequent experiments corroborated that ZLHXTY remarkably attenuated inflammation and oxidative stress damage in retinal tissues. Further experiments demonstrated that ZLHXTY inhibited the NLRP3/Caspase-1/GSDMD signaling pathway and related protein expression. Finally, TEM results also verified that ZLHXTY alleviated pyroptosis in retinal cells.
Our results suggest that ZLHXTY by regulating the NLRP3/Caspase-1/GSDMD axis, inhibiting pyroptosis, thereby relieving retinal dysfunction and vascular injury in HR mice.
蛭龙活血通瘀胶囊（ZLHXTY）已用于高血压所致血管疾病的临床治疗20多年。然而，ZLHXTY缓解高血压性视网膜病变（HR）的具体机制仍需进一步探索。
通过皮下植入渗透微型泵输注血管紧张素II建立HR小鼠模型，随后口服ZLHXTY（0.35、0.7、1.4 g/kg/天）进行28天治疗。为评估ZLHXTY对视网膜神经变性和血管损伤的影响，进行了光学相干断层扫描血管造影（OCTA）、视网膜电图（ERG）和苏木精-伊红（HE）染色等多项实验。随后，采用网络药理学和无标记4D蛋白质组学来阐明ZLHXTY缓解HR的潜在靶点和机制。最后，利用蛋白质免疫印迹法（Western blot）、酶联免疫吸附测定（ELISA）、免疫荧光（IF）等技术检测与炎症、氧化应激和NLRP3炎性小体激活相关的蛋白质表达。
ZLHXTY显著缓解了HR小鼠的视网膜功能障碍，增加了视网膜血流量，并减轻了视网膜组织水肿等病理变化。网络药理学表明，ZLHXTY可能通过肿瘤坏死因子（TNF）和核因子κB（NF-κB）等靶点发挥抗炎和抗氧化应激作用。蛋白质组学分析显示，ZL组和血管紧张素II组之间的差异蛋白主要富集于免疫炎症反应，其主要机制可能与NLRP3炎性小体的组装有关。随后实验证实，ZLHXTY显著减轻了视网膜组织中的炎症和氧化应激损伤。进一步实验表明，ZLHXTY抑制了NLRP3/半胱天冬酶-1（Caspase-1）/Gasdermin D（GSDMD）信号通路及相关蛋白表达。最后，透射电子显微镜（TEM）结果也证实ZLHXTY减轻了视网膜细胞的焦亡。
我们的结果表明，ZLHXTY通过调节NLRP3/Caspase-1/GSDMD轴，抑制焦亡，从而缓解HR小鼠的视网膜功能障碍和血管损伤。
