Zhang Wen-sheng, Li Feng, Bao Jun-qiang
Department of Traditional Chinese Medicine, Xijing Hospital, Fourth Military Medical University, Xi'an.
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2008 Sep;28(9):818-23.
To investigate the cathartic effect of total anthraquinone (AQ) from rhubarb on SD rats and its regulatory effect on aquaporin 4 (AQP4) expression in rat colon and in vitro cultured LoVo cell line.
Twenty-four SD rats were randomly divided into the normal control group treated with distilled water, and the two AQ groups administered with AQ suspension in cathartic and high dose (AQcd and AQhd) respectively via gastrogavage for 5 days. Water content in colonic stool was detected and the expression of AQP4 in rat's proximal colon was measured using Western blot and RT-PCR. LoVo cells cultured in vitro were used in the experimental study. The AQP4 protein and mRNA expressions in the cells were detected by Western blot and semiquantitative RT-PCR after they were cultured for 24 h with RPMI-1640 medium containing rhein/emodin in different concentrations, and those cultured with RPMI-1640 containing 20 mg/L rhein/emodin for different time points.
After treatment, the stool water content in the AQcd and AQhd groups was higher than that in the control group and the AQP4 expression in rats treated with AQ decreased in a dose-dependent manner. The study showed that rhein/emodin could significantly down-regulate the protein and mRNA expressions of AQP4 in cultured LoVo cells, with the effectiveness related with dose and acting time.
At the same time of playing cathartic action, total AQ of rhubarb can effectively down-regulate the expression of AQP4 in rat's proximal colon; rhein/emodin can suppress the AQP4 expression in LoVo cells in vitro. One mechanism of cathartic effect of rhubarb AQ is possibly its down-regulation on AQP4 expression.
探讨大黄总蒽醌(AQ)对SD大鼠的泻下作用及其对大鼠结肠和体外培养的LoVo细胞系中水通道蛋白4(AQP4)表达的调控作用。
将24只SD大鼠随机分为用蒸馏水治疗的正常对照组,以及分别通过灌胃给予泻下剂量和高剂量AQ悬浮液(AQcd和AQhd)的两个AQ组,连续给药5天。检测结肠粪便中的水分含量,并使用蛋白质免疫印迹法和逆转录-聚合酶链反应(RT-PCR)检测大鼠近端结肠中AQP4的表达。体外培养的LoVo细胞用于实验研究。在用不同浓度大黄酸/大黄素的RPMI-1640培养基培养24小时后,以及用含20mg/L大黄酸/大黄素的RPMI-1640培养基培养不同时间点后,通过蛋白质免疫印迹法和半定量RT-PCR检测细胞中AQP4蛋白和mRNA的表达。
治疗后,AQcd组和AQhd组的粪便水分含量高于对照组,且AQ处理的大鼠中AQP4表达呈剂量依赖性降低。研究表明,大黄酸/大黄素可显著下调培养的LoVo细胞中AQP4的蛋白和mRNA表达,其效果与剂量和作用时间有关。
大黄总AQ在发挥泻下作用的同时,可有效下调大鼠近端结肠中AQP4的表达;大黄酸/大黄素可在体外抑制LoVo细胞中AQP4的表达。大黄AQ泻下作用的一个机制可能是其对AQP4表达的下调。