Objective

Chinese herbal formulas show considerable therapeutic benefits in dementia. This study specifically explored the protective action of Zishen Huoxue recipe on the neurovascular unit (NVU) of rats with vascular dementia (VD).

Methods

VD rat models were established by permanent bilateral common carotid artery occlusion and treated with Zishen Huoxue recipe. In vitro glucose‑oxygen deprivation (OGD)-injured NVU models were established and treated with miR-124-3p agomir or rat medicated serum. The neurological damage, histopathological changes, and neuronal injury in the rat hippocampus were assessed using Morris water maze test and histological stainings. Expression of miR-124-3p was determined using RT-qPCR. The blood-brain barrier/NVU injury, cell pyroptosis, NLRP3 inflammasome activation, and release of inflammatory factors were analyzed mainly by immunofluorescence analysis, TUNEL staining, Western blot, and ELISA. QS-21 (an NLRP3 activator) was used to verify the role of miR-124-3p/NLRP3.

Results

Zishen Huoxue recipe ameliorated the learning/memory deficits, neuronal injury, NVU insults, cell pyroptosis, activation of NLRP3 inflammasome, and extensive secretion of lactate dehydrogenase/IL-1β/IL-18 in VD rats. miR-124-3p was downregulated in VD rats but upregulated after treatment of this recipe. miR-124-3p overexpression ameliorated NVU insults, reduced cell pyroptosis, lowered NLRP3 inflammasome activation, and suppressed inflammatory responses in OGD-injured NVU models. NLRP3 inflammasome activation partly counteracted the amelioration effect of miR-124-3p on pyroptosis. Zishen Huoxue recipe could upregulate miR-124-3p to suppress pyroptosis and protect NVU function.

Conclusion

Zishen Huoxue recipe can upregulate miR-124-3p expression to repress the inflammatory cascade-evoked pyroptosis, thereby protecting against neuronal injury in the NVU of VD rats.

中文翻译：

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滋肾活血方通过干扰炎症级联诱导的细胞焦亡对血管性痴呆大鼠神经血管单元神经元损伤的保护作用

客观的

中草药配方对痴呆症具有显着的治疗功效。本研究专门探讨滋肾活血方对血管性痴呆（VD）大鼠神经血管单位（NVU）的保护作用。

方法

采用双侧颈总动脉永久性闭塞法建立VD大鼠模型，并采用滋肾活血方治疗。建立体外葡萄糖缺氧（OGD）损伤的 NVU 模型，并用 miR-124-3p agomir 或大鼠含药血清进行治疗。采用Morris水迷宫试验和组织学染色评估大鼠海马的神经损伤、组织病理学变化和神经元损伤。使用 RT-qPCR 测定 miR-124-3p 的表达。主要通过免疫荧光分析、TUNEL染色、Western blot和ELISA等方法分析血脑屏障/NVU损伤、细胞焦亡、NLRP3炎症小体激活和炎症因子释放。QS-21（一种NLRP3激活剂）用于验证miR-124-3p/NLRP3的作用。

结果

滋肾活血方可改善 VD 大鼠的学习/记忆缺陷、神经元损伤、NVU 损伤、细胞焦亡、NLRP3 炎性体激活以及乳酸脱氢酶/IL-1β/IL-18 的广泛分泌。miR-124-3p 在 VD 大鼠中下调，但在该配方治疗后上调。在 OGD 损伤的 NVU 模型中，miR-124-3p 过表达可改善 NVU 损伤、减少细胞焦亡、降低 NLRP3 炎性体激活并抑制炎症反应。NLRP3 炎性体激活部分抵消了 miR-124-3p 对细胞焦亡的改善作用。滋肾活血方可上调miR-124-3p，抑制细胞焦亡，保护NVU功能。

结论

滋肾活血方可以上调miR-124-3p表达，抑制炎症级联反应诱发的细胞焦亡，从而保护VD大鼠NVU免受神经元损伤。