22-氧代胆钙化醇通过抑制 NF-κB/TNF-α 通路保护心肌缺血再灌注损伤。
22-oxacalcitriol protects myocardial ischemia-reperfusion injury by suppressing NF-κB/TNF-α pathway.
机构信息
Department of Public Health, Yantaishan Hospital, Yantai, China.
出版信息
Eur Rev Med Pharmacol Sci. 2019 Jun;23(12):5495-5502. doi: 10.26355/eurrev_201906_18219.
OBJECTIVE
The aim of this study was to explore whether 22-oxacalcitriol could protect inflammatory response induced by ischemia-reperfusion injury (IRI) in rats, and to investigate its underlying mechanism.
MATERIALS AND METHODS
24 male Sprague Dawley rats were randomly assigned into the sham group, the IRI group and the 22-oxacalcitriol group, with 8 rats in each group. Serum and heart samples of each rat were collected 10 days after the animal procedure. The serum levels of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) in each rat were detected by relative commercial kits. Pathological lesions in rat myocardium were observed by hematoxylin and eosin (HE) staining. Cardiomyocyte apoptosis in rat heart was accessed by TUNEL staining. Meanwhile, the serum levels of tumor necrosis factor-α (TNF-α), interleukin 1 beta (IL-1β), interleukin-6 (IL-6), and KC-GRO were detected by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR). Also, the protein expression levels of NF-κB, TNF-α, VCAM-1, ICAM-1, and MCP-1 in rat myocardium were detected by Western blot and immunohistochemistry.
RESULTS
The serum levels of CK-MB and LDH in rats of the IRI group were significantly higher than those of the sham group. 22-oxacalcitriol treatment remarkably decreased the serum levels of CK-MB and LDH when compared with the IRI group. However, cardiomyocyte apoptosis of the 22-oxacalcitriol group was markedly less than the IRI group. The activities of SOD, GSH, CAT and T-AOC in the cardiac homogenate of the 22-oxacalcitriol group were significantly elevated than those of the IRI group. Meanwhile, malondialdehyde (MDA) and reactive oxygen species (ROS) levels were remarkably decreased by 22-oxacalcitriol treatment. Furthermore, the serum levels of TNF-α, IL-1β, IL-6 and KC-GRO were significantly downregulated in the 22-oxacalcitriol group. Western blot results showed that the protein expression levels of NF-κB, TNF-α, VCAM-1, ICAM-1 and MCP-1 in the 22-oxacalcitriol group were significantly lower than those of the IRI group.
CONCLUSIONS
22-oxacalcitriol inhibits the inflammatory response in the myocardium by suppressing NF-kB/TNF-α pathway, thereby protecting myocardial ischemia-reperfusion injury in rats.
目的
本研究旨在探讨 22-氧代钙三醇是否能减轻大鼠缺血再灌注损伤（IRI）引起的炎症反应，并探讨其作用机制。
材料和方法
将 24 只雄性 Sprague Dawley 大鼠随机分为假手术组、IRI 组和 22-氧代钙三醇组，每组 8 只。在动物手术后 10 天，收集每组大鼠的血清和心脏样本。通过相对商业试剂盒检测大鼠血清肌酸激酶同工酶（CK-MB）和乳酸脱氢酶（LDH）水平。苏木精和伊红（HE）染色观察大鼠心肌病理损伤。TUNEL 染色检测大鼠心肌细胞凋亡。同时，通过实时定量聚合酶链反应（RT-qPCR）检测血清肿瘤坏死因子-α（TNF-α）、白细胞介素 1β（IL-1β）、白细胞介素-6（IL-6）和 KC-GRO 的水平。Western blot 和免疫组织化学检测大鼠心肌组织中 NF-κB、TNF-α、VCAM-1、ICAM-1 和 MCP-1 的蛋白表达水平。
结果
IRI 组大鼠血清 CK-MB 和 LDH 水平明显高于假手术组。与 IRI 组相比，22-氧代钙三醇治疗显著降低了血清 CK-MB 和 LDH 水平。然而，22-氧代钙三醇组的心肌细胞凋亡明显少于 IRI 组。22-氧代钙三醇组心脏匀浆中 SOD、GSH、CAT 和 T-AOC 的活性明显高于 IRI 组。同时，22-氧代钙三醇治疗可显著降低丙二醛（MDA）和活性氧（ROS）水平。此外，22-氧代钙三醇组血清 TNF-α、IL-1β、IL-6 和 KC-GRO 水平明显下调。Western blot 结果显示，22-氧代钙三醇组 NF-κB、TNF-α、VCAM-1、ICAM-1 和 MCP-1 的蛋白表达水平明显低于 IRI 组。
结论
22-氧代钙三醇通过抑制 NF-κB/TNF-α 通路抑制心肌炎症反应，从而保护大鼠心肌缺血再灌注损伤。
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