Department of Pharmacology, Faculty of Medicine, Kastamonu University, 3700, Kastamonu, Turkey.
Faculty of Medicine, Department of Pharmacology, Kafkas University, Kars, Turkey.
Cardiovasc Toxicol. 2022 Jan;22(1):52-62. doi: 10.1007/s12012-021-09698-3. Epub 2021 Oct 1.
The aim of the study was to examine the protective effects and possible mechanism of gossypin against isoproterenol (ISO)-mediated myocardial damage in vivo and H9c2 cell damage in vitro. H9c2 cells were categorized into five groups. Viability was evaluated with MTT and LDH release in H9c2 cells. Apoptotic parameter analysis was performed with cytochrome c (Cyt-c), caspase-3 (CASP-3), and BCL2/Bax mRNA expression levels. In vivo, gossypin was administered orally to mice at doses of 5, 10, and 20 mg/kg for 7 days. ISO groups were injected with isoproterenol (150 mg/kg) subcutaneously (on 8th and 9th) for 2 days. Afterward, lactate dehydrogenase (LDH), creatine kinase-MB (CK-MB) levels and Troponin-I (Tn-I) amount from their serum, oxidative stress parameters superoxide dismutase (SOD) activity, glutathione (GSH) and malondialdehyde (MDA) levels, and tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1 β), and NF-kB mRNA expression levels with inflammatory markers from heart tissue were evaluated. In addition, IL-1B, BCL-2, and cas-3 immunohistochemical staining was performed from heart tissue and TNF-a level was measured by ELISA method. Administration of Gossypin protected the cells by dose-dependent, eliminating the reduced cell viability and increased LDH release of ISO in H9c2 cells. In mice serum analyses, increased LDH, CK-MB levels, and Tn-I levels were normalized by gossypin. ISO administration in heart tissue is regulated by gossypin with increased SOD activity, GSH amount, TNF-α, IL-6, IL-1β, and NF-kB mRNA expression levels and decreased MDA amount. Overall, the present results demonstrated that gossypin has a potential cardioprotective treatment for ischemic heart disease on in vivo and in vitro.
本研究旨在探讨棉花素对体内异丙肾上腺素（ISO）介导的心肌损伤和体外 H9c2 细胞损伤的保护作用及其可能机制。将 H9c2 细胞分为五组。通过 MTT 和 LDH 释放评估 H9c2 细胞活力。通过细胞色素 c（Cyt-c）、半胱天冬酶-3（CASP-3）和 BCL2/Bax mRNA 表达水平分析凋亡参数。体内，棉花素以 5、10 和 20mg/kg 的剂量口服给药，持续 7 天。ISO 组在第 8 和第 9 天皮下注射异丙肾上腺素（150mg/kg），持续 2 天。随后，评估其血清中乳酸脱氢酶（LDH）、肌酸激酶-MB（CK-MB）水平和肌钙蛋白-I（Tn-I）含量、氧化应激参数超氧化物歧化酶（SOD）活性、谷胱甘肽（GSH）和丙二醛（MDA）水平，以及肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）和核因子-κB（NF-kB）mRNA 表达水平以及心脏组织中的炎症标志物。此外，还进行了心脏组织中 IL-1B、BCL-2 和 cas-3 的免疫组织化学染色，并通过 ELISA 法测量 TNF-a 水平。棉花素通过剂量依赖性方式保护细胞，消除 ISO 对 H9c2 细胞活力的降低和 LDH 释放的增加。在小鼠血清分析中，棉花素使 LDH、CK-MB 水平和 Tn-I 水平升高得到正常化。棉花素对心脏组织中 ISO 的作用受调节，SOD 活性、GSH 含量、TNF-α、IL-6、IL-1β 和 NF-kB mRNA 表达水平升高，MDA 含量降低。总的来说，这些结果表明棉花素具有体内和体外缺血性心脏病的潜在心脏保护治疗作用。
