Department of Vascular Surgery, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing, 210008, China.
Jiangsu Key Laboratory of Molecular Medicine, Medical School of Nanjing University, Nanjing, 210093, China.
J Ethnopharmacol. 2022 Jun 28;292:115206. doi: 10.1016/j.jep.2022.115206. Epub 2022 Mar 14.
Oridonin (Ori), extracted from Isodon rubescens (Hemsl.) H.Hara, is a well-known traditional Chinese herbal medicinal product that possesses antioxidant and anti-inflammatory activities. Oxidative stress and inflammation are the main pathophysiological mechanisms in hindlimb IR injury. However, whether Ori has a protective effect on hind limb IR injury is unknown.
The present study was designed to determine the effect of Ori on hindlimb IR injury and its relationship with oxidative stress and inflammation.
The hind limb IR injury model in mice was used to evaluate the protective effect and related mechanisms of Ori. Forty-eight C57BL/6 mice (n = 12 per group) were randomly divided into four groups: Sham group; IR group; IR + Ori (10 mg/kg) group and IR + Ori (20 mg/kg) group. Mice in the IR and IR + Ori groups were subjected to hindlimb IR injury, while mice in the Sham group were subjected to no hindlimb IR injury. HE staining, Masson's staining, TTC staining, DHE staining, TUNEL staining, western blotting analysis and quantitative real-time PCR were employed to explore the mechanisms by which Ori exerts a protective effect on a classical hindlimb IR model in mice.
We found that Ori pretreatment prevented muscle damage and decreased cell apoptosis levels compared with the vehicle control. Moreover, the SOD2, CAT, MDA and ROS levels in muscle showed that Ori could significantly reduce oxidative stress in hindlimb IR mice, while the IL-1β and TNF-α levels in muscle showed that Ori could significantly attenuate IR-induced inflammation. We also found that Ori could increase the expression of Nrf2 and its downstream protein HO-1 and inhibit the expression levels of NLRP3-related proteins (NLRP3, ASC and Caspase-1) in vivo.
Our study suggested that Ori has a protective effect on hindlimb IR injury, which may be related to Nrf2-mediated oxidative stress and NLRP3-mediated inflammasome activation.
冬凌草甲素（Ori）是从冬凌草（Isodon rubescens（Hemsl.）H.Hara）中提取的一种著名的传统中药，具有抗氧化和抗炎作用。氧化应激和炎症是下肢缺血再灌注损伤的主要病理生理机制。然而，冬凌草甲素（Ori）是否对下肢缺血再灌注损伤具有保护作用尚不清楚。
本研究旨在探讨冬凌草甲素（Ori）对下肢缺血再灌注损伤的作用及其与氧化应激和炎症的关系。
采用小鼠下肢缺血再灌注损伤模型评价冬凌草甲素（Ori）的保护作用及相关机制。将 48 只 C57BL/6 小鼠（每组 12 只）随机分为 4 组：假手术组；缺血再灌注组；缺血再灌注+冬凌草甲素（10mg/kg）组；缺血再灌注+冬凌草甲素（20mg/kg）组。缺血再灌注组和缺血再灌注+冬凌草甲素（Ori）组小鼠进行下肢缺血再灌注损伤，假手术组小鼠不进行下肢缺血再灌注损伤。采用 HE 染色、Masson 染色、TTC 染色、DHE 染色、TUNEL 染色、Western blot 分析和实时定量 PCR 等方法探讨冬凌草甲素（Ori）对经典小鼠下肢缺血再灌注模型发挥保护作用的机制。
我们发现，与载体对照组相比，冬凌草甲素（Ori）预处理可防止肌肉损伤并降低细胞凋亡水平。此外，肌肉中的 SOD2、CAT、MDA 和 ROS 水平表明，冬凌草甲素（Ori）可显著减轻下肢缺血再灌注小鼠的氧化应激，而肌肉中的 IL-1β和 TNF-α水平表明，冬凌草甲素（Ori）可显著抑制 IR 诱导的炎症。我们还发现，冬凌草甲素（Ori）可增加 Nrf2 及其下游蛋白 HO-1 的表达，并抑制体内 NLRP3 相关蛋白（NLRP3、ASC 和 Caspase-1）的表达水平。
本研究表明，冬凌草甲素（Ori）对下肢缺血再灌注损伤具有保护作用，这可能与 Nrf2 介导的氧化应激和 NLRP3 介导的炎症小体激活有关。
