砂仁水提物对5-FU致大鼠肠黏膜屏障损伤的保护作用

Protective Effect of Amomi Fructus Water Extraction on Intestinal Mucositis Induced by 5-FU

关键词:  砂仁水提物  肠黏膜屏障  5-氟尿嘧啶  炎症

Key words:  Amomi Fructus water extraction  intestinal mucosal barrier  5-FU  inflammation

摘要:

目的 研究砂仁水提物对5-氟尿嘧啶（5-FU）诱导的大鼠肠黏膜损伤的保护作用。方法 清洁级SD大鼠随机分成正常对照组、模型组、阳性药组、砂仁水提物48，96，192 mg·kg^-1组，每组8只。正常对照组腹腔注射生理盐水5 d，其余各组腹腔注射35 mg·kg^-1 5-FU 5 d，且从实验开始正常对照组空腹灌胃生理盐水、其余各组空腹灌胃相应治疗药物，给药12 d。实验期间记录大鼠腹泻情况、体质量、食量；实验结束后取小肠做病理切片；ELISA法检测大鼠IL-6、ROS、NF-κB、MPO、TNF-α、LPS的含量，流式细胞术检测肠道紧密连接蛋白ZO-1、occludin及caspase-3的含量。结果 砂仁水提物可显著改善5-FU导致的大鼠体质量、食量下降；显著改善5-FU诱导的大鼠肠屏障损伤及腹泻；显著降低大鼠血液中炎症因子IL-6、ROS的含量；显著抑制5-FU导致的大鼠小肠中NF-κB、TNF-α、MPO的升高与阳性药效果相当。同时砂仁水提物可增加occludin、抑制caspase-3的表达，在一定程度上抑制肠源性LPS易位，保护肠屏障功能。结论 砂仁水提物对5-FU引起的肠黏膜屏障损伤具有一定保护作用，其中96 mg·kg^-1的效果较好，推测其作用机制可能与砂仁水提物抑制肠黏膜炎前体物质ROS的产生，从而抑制核因子NF-κB通路，降低炎症反应；抑制MPO的激活及caspase-3表达，维持细胞间紧密完整性进而抑制肠源性LPS易位有关。

Abstract:

OBJECTIVE To study the effect of Amomi Fructus water extraction on intestinal mucosal barrier function in rats induced by 5-FU. METHODS SPF SD rats were divided into normal control group, model group, positive drug group and water extract of Amomi Fructus(48, 96, 192 mg·kg^-1) groups, 8 rats in each group. The normal control group was intraperitoneally injected with saline for 5 d, and the other groups were intraperitoneally injected with 35 mg·kg^-1 5-FU for 5 d. From the beginning of the experiment, the normal control group were given saline, and the other groups were given the corresponding therapeutic drugs for fasting and gavage for 12 d. Diarrhea occurrence, weight and food intakes were recorded daily throughout the study in rats. Blood samples and the small intestinal were collected after killing of rats for evaluation of histopathology, the content of cytokines (IL-6, ROS, NF-kB, MPO and TNF-α) and LPS were detected by ELISA. The intestinal tight junction protein ZO-1, occludin and caspase-3 were measured by flow cytometry. RESULTS Water extract of Amomi Fructus significantly improved rats body weight, food intakes diarrhea and significantly reversed the hisopathological changes in intestinal mucositis induced by 5-FU and reduced the inflammatory parameters (IL-6, ROS) in blood sample. It could also significantly effectively reduce the content of NF-κB, TNF-α, MPO in small intestinal tissue and the effect was similar to the positive drug. Meanwhile, it showed that water extract of Amomi Fructus could increase the expression of occludin and decrease the expression of caspase-3, inhibit the translocation of LPS, protect the integrity of intestinal barrier function. CONCLUSION It is suggested that the water extract of Amomi Fructus may play an important protective role in intestinal mucositis and gut barrier injury in rats challenged by 5-FU, 96 mg·kg^-1 shows best effect, which may contribute to inhibit ROS generated, decrease pro-inflammatory cytokines secrete, inhibit NF-κB signaling, caspase-3 signaling and suppress the MPO activity, further decrease apoptosis, inhibit the translocation of LPS and protect the integrity of mucosal.