Graduate School, Beijing University of Chinese Medicine, Beijing 100029, China; Department of Emergency, China-Japan Friendship Hospital, Beijing 100029, China.
Department of Emergency, China-Japan Friendship Hospital, Beijing 100029, China; Graduate School of Peking Union Medical College, Beijing 100730, China.
Int Immunopharmacol. 2023 May;118:110049. doi: 10.1016/j.intimp.2023.110049. Epub 2023 Apr 3.
Sepsis has complex pathophysiological mechanisms that bring new challenges in the treatment of sepsis at a time when the intestinal microcirculation in sepsis is receiving increasing attention. Dl-3-n-butylphthalide (NBP), which is a drug that can improve multiorgan ischemic diseases, is also worth examining to improve the intestinal microcirculation in sepsis.
In this study, male Sprague-Dawley rats were divided into the sham group (n = 6), CLP group (n = 6), NBP group (n = 6) and NBP + LY294002 group (n = 6). The rat model of severe sepsis was established by cecal ligation and puncture (CLP). Abdominal wall incisions and sutures were performed in the first group, and CLP was performed in the latter three groups. Normal saline/NBP/NBP + LY294002 solution was injected intraperitoneally 2 h or 1 h before modeling. Hemodynamic data (blood pressure and heart rate) were recorded at 0, 2, 4 and 6 h. Sidestream dark field (SDF) imaging and the Medsoft System were used to observe the intestinal microcirculation of rats and obtain data at 0, 2, 4, and 6 h. Six hours after the model was established, the serum levels of TNF-α and IL-6 were measured to evaluate the level of systemic inflammation. Pathological damage to the small intestine was evaluated by electron microscopy and histological analysis. The expression levels of P-PI3K, PI3K, P-AKT, AKT, LC3 and p62 in the small intestine were analyzed by Western blotting. The expressions of P-PI3K, P-AKT, LC3 and P62 in small intestine were detected by immunohistochemical staining.
NBP improved intestinal microcirculation disturbances in septic rats, alleviated the systemic inflammatory response, reduced the destruction of the small intestinal mucosa and the disruption of microvascular endothelial cells, and alleviated autophagy in vascular endothelial cells. NBP increased the ratio of P-PI3K/total PI3K, P-AKT/total AKT, and P62/β-actin and decreased the ratio of LC3 II/LC3 I.
NBP ameliorated intestinal microcirculation disturbances and the destruction of small intestinal vascular endothelial cells in septic rats by activating the PI3K/Akt signaling pathway and regulating autophagy.
脓毒症具有复杂的病理生理学机制，在肠道微循环越来越受到重视的情况下，脓毒症的治疗也带来了新的挑战。能够改善多器官缺血性疾病的药物 3-正丁基苯酞（NBP）也值得研究，以改善脓毒症的肠道微循环。
本研究将雄性 Sprague-Dawley 大鼠分为假手术组（n=6）、CLP 组（n=6）、NBP 组（n=6）和 NBP+LY294002 组（n=6）。通过盲肠结扎和穿孔（CLP）建立严重脓毒症大鼠模型。第一组进行腹壁切开和缝合，后三组进行 CLP。造模前 2 h 或 1 h 经腹腔注射生理盐水/NBP/NBP+LY294002 溶液。在 0、2、4 和 6 h 记录血流动力学数据（血压和心率）。采用边流暗场（SDF）成像和 Medsoft 系统观察大鼠肠道微循环并获得 0、2、4 和 6 h 的数据。模型建立后 6 h，检测血清 TNF-α和 IL-6 水平评估全身炎症反应水平。电镜和组织学分析评估小肠的病理损伤。通过 Western blot 分析小肠中 P-PI3K、PI3K、P-AKT、AKT、LC3 和 p62 的表达水平。免疫组化染色检测小肠中 P-PI3K、P-AKT、LC3 和 P62 的表达。
NBP 改善了脓毒症大鼠肠道微循环障碍，减轻了全身炎症反应，减少了小肠黏膜的破坏和微血管内皮细胞的破坏，并减轻了血管内皮细胞的自噬。NBP 增加了 P-PI3K/总 PI3K、P-AKT/总 AKT 和 P62/β-肌动蛋白的比值，降低了 LC3 II/LC3 I 的比值。
NBP 通过激活 PI3K/Akt 信号通路和调节自噬，改善脓毒症大鼠肠道微循环障碍和小肠血管内皮细胞的破坏。
