Türkcü Gül, Alabalık Ulaş, Keleş Ayşe Nur, Bozkurt Mehtap, İbiloğlu İbrahim, Fırat Uğur, Büyükbayram Hüseyin
Department of Pathology, Dicle University Faculty of Medicine Diyarbakir, Turkey.
Department of Physical Medicine and Rehabilitation, Dicle University Faculty of Medicine Diyarbakir, Turkey.
Int J Clin Exp Med. 2015 Sep 15;8(9):15474-81. eCollection 2015.
The purpose of this experimental study was to evaluate the efficacy of carvacrol (CVR) and pomegranate (PMG) against methotrexate (MTX)-induced intestinal damage using histopathological and immunohistochemical techniques.
Thirty-two male Sprague-Dawley rats, weighing 195-250 g, were divided into four groups: control, MTX treatment alone, MTX plus CVR and MTX plus PMG. A single dose of CVR (73 mg/kg) was administered intraperitoneally to group III on the first day of the experiment, PMG (225 mg/kg/day) was administered orogastrically (with a gavage needle) once daily for 7 days and a single dose of MTX (20 mg/kg) was administered intraperitoneally on the second day of the experiment. Intestinal tissues were obtained on 8(th) day, and examined for villus damage, crypt damage, and inflammation. Ki-67 and Caspase 3 staining was used for immunohistochemical evaluation.
MTX treatment induced villus shortening and fusion, epithelial atrophy, crypt loss, inflammatory infiltrate in the lamina propria, and goblet cell depletion. The CVR and PMG decreased the severity of intestinal damage caused by MTX treatment. In the MTX-received group, significant inflammatory cell infiltration was observed in the lamina propria. Compared to the MTX-received group, the PMG and CVR groups showed less villus and crypt damage and less inflammation in the lamina propria. Fewer Ki-67 positive cells were observed in the crypts of the MTX-received groups compared to the control group. There were more Ki-67 positive cells in the CVR and PMG groups compared to MTX group. The MTX-received group exhibited more caspase-3 positive cells than the control group, and the number of caspase-3 positive cells were decreased in the CVR and PMG treated groups.
This study is the first to show that PMG and CVR decrease MTX-related damage and apoptotic activity in intestinal tissue.
本实验研究旨在运用组织病理学和免疫组织化学技术，评估香芹酚（CVR）和石榴（PMG）对甲氨蝶呤（MTX）诱导的肠道损伤的疗效。
将32只体重195 - 250克的雄性Sprague-Dawley大鼠分为四组：对照组、单独MTX治疗组、MTX加CVR组和MTX加PMG组。实验第一天，向第三组腹腔注射单剂量的CVR（73毫克/千克），向PMG组每日经口胃管（用灌胃针）给予PMG（225毫克/千克/天），持续7天，实验第二天腹腔注射单剂量的MTX（20毫克/千克）。在第8天获取肠道组织，检查绒毛损伤、隐窝损伤和炎症情况。采用Ki-67和半胱天冬酶3染色进行免疫组织化学评估。
MTX治疗导致绒毛缩短和融合、上皮萎缩、隐窝缺失、固有层炎症浸润以及杯状细胞减少。CVR和PMG减轻了MTX治疗引起的肠道损伤严重程度。在接受MTX的组中，固有层观察到明显的炎性细胞浸润。与接受MTX的组相比，PMG组和CVR组的绒毛和隐窝损伤较轻，固有层炎症较少。与对照组相比，接受MTX的组隐窝中观察到的Ki-67阳性细胞较少。与MTX组相比，CVR组和PMG组的Ki-67阳性细胞更多。接受MTX的组比对照组表现出更多的半胱天冬酶-3阳性细胞，而CVR组和PMG治疗组的半胱天冬酶-3阳性细胞数量减少。
本研究首次表明，PMG和CVR可减少肠道组织中MTX相关的损伤和凋亡活性。
