补阳还五汤苷类成分对脑缺血再灌注损伤大鼠细胞焦亡的神经保护作用。

Neuroprotective effect of glycosides in Buyang Huanwu Decoction on pyroptosis following cerebral ischemia-reperfusion injury in rats.

机构信息

Laboratory of Vascular Biology, Medical College, Hunan University of Chinese Medicine, Changsha, 410208, China.

The Third Affiliated Hospital of Hunan University of Chinese Medicine, Zhuzhou, 412000, China.

出版信息

J Ethnopharmacol. 2019 Oct 5;242:112051. doi: 10.1016/j.jep.2019.112051. Epub 2019 Jul 3.

DOI:10.1016/j.jep.2019.112051

PMID:31279072

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Buyang Huanwu Decoction (BYHWD) is used in classical traditional Chinese medicine to prevent and treat cerebral ischemia. Glycosides, which are effective components extracted from BYHWD, mainly include astragaloside IV, paeoniflorin, and amygdalin. These glycosides are the primary pharmacologically effective constituents of BYHWD that act against cerebral ischemic nerve injury; however, the mechanism of action of BYHWD is still unclear.

AIM OF THE STUDY

The present study aimed to determine the effect of BYHWD glycosides on pyroptosis after cerebral ischemia reperfusion injury and explore whether its mechanism involves the classical pyroptosis pathway mediated by NLRP3.

MATERIAL AND METHODS

Adult male Sprague-Dawley rats (n = 140) were randomly divided into seven groups: sham, cerebral ischemia and reperfusion (I/R), glycosides (0.064 g/kg, 0.128 g/kg, and 0.256 g/kg), BYHWD, and AC-YVAD-CMK (caspase-1 inhibitor). A rat model of cerebral I/R was established via classic middle cerebral artery occlusion (MCAO) for 2 h, followed by 24-h reperfusion. Neurological function was estimated using neurological defect scores. Brain infarct volumes were determined by 2,3,5-triphenyltetrazolium chloride (TTC) staining, and nerve cell damage was evaluated by Nissl staining. Pyroptosis was detected using TUNEL and caspase-1 immunofluorescence double staining. Protein expression of NLRP3, ASC, caspase-1, pro-caspase-1, and IL-1β was analyzed using Western blot analysis.

RESULTS

Glycosides improved neurological dysfunction, alleviated neuronal damage, and inhibited neuronal pyroptosis. The 0.128 g/kg glycosides group showed the most significant effects. Furthermore, we observed that this group showed significant inhibition of the expression of NLRP3, ASC, pro-caspase-1, caspase-1, and IL-1β proteins of the NLRP3-mediated classical pathway of pyroptosis.

CONCLUSIONS

Glycosides exert neuroprotective effects by inhibiting pyroptosis of neurons after cerebral I/R injury. The underlying mechanism of action is closely related to the regulation of the classical pyroptosis pathway by NLRP3.

摘要

民族药理学相关性

补阳还五汤（BYHWD）在经典的中医中被用于预防和治疗脑缺血。从 BYHWD 中提取的糖苷类化合物是其主要的有效成分，包括黄芪甲苷、芍药苷和苦杏仁苷。这些糖苷类化合物是 BYHWD 对抗脑缺血性神经损伤的主要药效成分，但 BYHWD 的作用机制尚不清楚。

研究目的

本研究旨在探讨 BYHWD 糖苷类化合物对脑缺血再灌注损伤后细胞焦亡的影响，并探讨其是否通过 NLRP3 介导的经典焦亡途径发挥作用。

材料和方法

成年雄性 Sprague-Dawley 大鼠（n=140）随机分为 7 组：假手术组、脑缺血再灌注组（I/R 组）、糖苷组（0.064 g/kg、0.128 g/kg 和 0.256 g/kg）、BYHWD 组和 AC-YVAD-CMK（caspase-1 抑制剂）组。采用经典大脑中动脉闭塞（MCAO）法建立大鼠脑 I/R 模型，缺血 2 h 后再灌注 24 h。用神经功能缺损评分评估神经功能。用 2,3,5-三苯基氯化四氮唑（TTC）染色法测定脑梗死体积，用尼氏染色法评估神经细胞损伤。用 TUNEL 和 caspase-1 免疫荧光双重染色法检测细胞焦亡。用 Western blot 分析法检测 NLRP3、ASC、caspase-1、pro-caspase-1 和 IL-1β 蛋白的表达。