Investigation of the role of rosmarinic acid treatment in regulating inflammation, cell damage, and angiogenesis in rat ovarian torsion and detorsion models

Abstract Purpose To investigate the protective effect of rosmarinic acid (RA) in ovarian ischemia/reperfusion injury using biochemical, histopathological, and immunohistochemical methods. Methods Wistar female rats (n = 32) were randomly divided into four groups: control, ischemia, ischemia-reperfusion, and ischemia-reperfusion with RA. Rosmarinic acid was given at a dose of 50 mg/kg by oral gavage three hours after reperfusion. Malondialdehyde (MDA) levels and glutathione peroxidase (GSH-Px) activities were determined in the ovary tissue homogenates for each rat. Results In the ischemia-reperfusion with RA group, the epithelial cells are regularly regulated at the periphery, and the degenerative changes in preantral and antral follicle cells are reduced. Follicle cells and cells in the corpus luteum showed a decrease in vascular endothelial growth factor (VEGF) expression, while VEGF demonstrated a positive reaction in vascular endothelial cells and stromal cells. The TNF-α expression due to the decreased degenerative effect and inflammation was positive in the macrophage cells. The expression of caspase-3 as an apoptosis change was negative in antral follicle cells and granular cells around the antral follicle. Conclusion Different doses of RA may be useful in preventing ischemic damage after vascularization, inflammation, and apoptotic development after ischemia/reperfusion.

摘要
研究目的：采用生化、组织病理及免疫组织化学方法，探究迷迭香酸（rosmarinic acid, RA）对卵巢缺血再灌注损伤的保护作用。
方法：将32只Wistar雌性大鼠随机分为4组：对照组、缺血组、缺血再灌注组以及迷迭香酸干预缺血再灌注组。于再灌注3小时后，以50 mg/kg的剂量通过灌胃方式给予迷迭香酸。对每只大鼠的卵巢组织匀浆进行丙二醛（Malondialdehyde, MDA）水平及谷胱甘肽过氧化物酶（glutathione peroxidase, GSH-Px）活性检测。
结果：在迷迭香酸干预缺血再灌注组中，上皮细胞周边排列规整，窦前卵泡与窦状卵泡细胞的退行性改变有所减轻。卵泡细胞与黄体细胞的血管内皮生长因子（vascular endothelial growth factor, VEGF）表达量降低，而血管内皮细胞及基质细胞中的VEGF呈阳性反应。因退行性改变与炎症反应得到缓解，巨噬细胞中的肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）表达呈阳性。作为细胞凋亡标志物的半胱氨酸天冬氨酸蛋白酶-3（caspase-3）在窦状卵泡细胞及窦状卵泡周围的颗粒细胞中表达呈阴性。
结论：不同剂量的迷迭香酸或可用于预防血管化后缺血损伤、炎症反应以及缺血再灌注后的细胞凋亡进展。