Department of Cardiology, Beijing Anzhen Hospital, Capital Medical University, Beijing 100020, P.R. China.
Changchun University of Chinese Medicine, Changchun, Jilin 130117, P.R. China.
Mol Med Rep. 2022 Oct;26(4). doi: 10.3892/mmr.2022.12834. Epub 2022 Aug 25.
Morphine is the most common drug of choice in clinical pain management; however, morphine tolerance presents a significant clinical challenge. The pathogenesis of morphine tolerance is known to be closely associated with angiotensin II receptor type 1 (AT1R) in microglia. As an AT1R antagonist, candesartan may serve an important role in regulating morphine tolerance. Therefore, the present study aimed to investigate the role of candesartan in morphine tolerance, and to explore the underlying mechanism. To meet this aim, BV2 microglial cells were treated with morphine or candesartan alone, or as a combination, and the expression levels of AT1R in BV2 cells were detected by reverse transcription‑quantitative PCR (RT‑qPCR) and western blotting. The levels of the inflammatory cytokines tumor necrosis factor‑α, interleukin (IL)‑1β and IL‑6 were subsequently detected by ELISA and western blotting. In addition, immunofluorescence analysis, western blotting and RT‑qPCR were used to detect the expression levels of the BV2 cell activation marker, ionized calcium‑binding adaptor molecule 1 (IBA‑1). Western blotting was also used to detect the expression levels of peroxisome proliferator‑activated receptor‑γ/AMP‑activated protein kinase (PPARγ/AMPK) signaling pathway‑associated proteins. Finally, the cells were treated with the PPARγ antagonist GW9662 and the AMPK inhibitor compound C to further explore the mechanism underlying the effects of candesartan on improving morphine tolerance. The expression levels of AT1R were revealed to be significantly increased following morphine induction; however, candesartan treatment inhibited the expression levels of AT1R, the levels of inflammatory cytokines and the protein expression levels of IBA‑1 in morphine‑induced BV2 cells in a dose‑dependent manner. These processes may be associated with activation of the PPARγ/AMPK signaling pathway. Taken together, the present study revealed that treatment with candesartan reduced morphine‑induced inflammatory response and cellular activation of BV2 cells via PPARγ/AMPK signaling.
吗啡是临床疼痛管理中最常用的药物选择；然而，吗啡耐受是一个重大的临床挑战。已知吗啡耐受的发病机制与小胶质细胞中的血管紧张素 II 受体 1（AT1R）密切相关。作为 AT1R 拮抗剂，坎地沙坦可能在调节吗啡耐受方面发挥重要作用。因此，本研究旨在探讨坎地沙坦在吗啡耐受中的作用，并探讨其潜在机制。为了达到这一目的，用吗啡或坎地沙坦单独或联合处理 BV2 小胶质细胞，并通过逆转录-定量 PCR（RT-qPCR）和 Western blot 检测 BV2 细胞中 AT1R 的表达水平。随后通过 ELISA 和 Western blot 检测炎症细胞因子肿瘤坏死因子-α、白细胞介素（IL）-1β和 IL-6 的水平。此外，通过免疫荧光分析、Western blot 和 RT-qPCR 检测 BV2 细胞激活标志物离子钙结合接头分子 1（IBA-1）的表达水平。Western blot 还用于检测过氧化物酶体增殖物激活受体-γ/AMP 激活蛋白激酶（PPARγ/AMPK）信号通路相关蛋白的表达水平。最后，用 PPARγ 拮抗剂 GW9662 和 AMPK 抑制剂化合物 C 处理细胞，进一步探讨坎地沙坦改善吗啡耐受作用的机制。结果表明，吗啡诱导后 AT1R 的表达水平明显增加；然而，坎地沙坦处理以剂量依赖性方式抑制吗啡诱导的 BV2 细胞中 AT1R 的表达水平、炎症细胞因子的水平和 IBA-1 的蛋白表达水平。这些过程可能与 PPARγ/AMPK 信号通路的激活有关。综上所述，本研究表明，坎地沙坦通过 PPARγ/AMPK 信号通路降低吗啡诱导的 BV2 细胞炎症反应和细胞激活。
