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目的利用正交试验结合体外模型测定固公果根总三萜(TTROG)提取物抑制大鼠离体肠平滑肌收缩活性,优化其提取工艺。方法以熊果酸为对照品,采用香草醛-冰醋酸显色,紫外分光光度法测定TTROG提取物中三萜含量,单因素考察提取溶剂、料液比、提取温度、提取时间、提取次数对TTROG提取率的影响;在单因素实验的基础上采用正交试验法考察提取溶剂、料液比、提取时间对TTROG提取率的影响;通过张力传感器与BL-420生物实验多通道生理信号采集处理系统记录不同提取物对体外肠平滑肌收缩的抑制作用,利用加权方法使生物活性与提取率相结合评价TTROG的提取工艺。结果 TTROG的最佳提取工艺条件为:提取溶剂80%乙醇,料液比1∶10,提取时间2 h,提取温度80℃,提取次数3次;最优条件下TTROG的提取率可达42.12 mg/g;TTROG提取物对十二指肠、空肠、回肠、结肠4个肠段都具有显著的抑制离体肠平滑肌收缩的作用,呈剂量相关性,对空肠的作用最强,抑制率可达41.96%。结论经优化后的TTROG提取工艺提取率高、稳定,且获取的TTROG提取物具有显著的抑制大鼠离体肠平滑肌收缩功能。
OBJECTIVE: To determine the contractile activity of rat intestinal smooth muscle in vitro by orthogonal test combined with in vitro model, and to optimize the extraction process. Methods ursolic acid as reference substance, using vanillin - glacial acetic acid colorimetric determination of triterpene content of TTROG extract, single factor to investigate the extraction solvent, solid-liquid ratio, extraction temperature, extraction time, extraction times TTROG extraction rate on the basis of single factor experiments using orthogonal test method to investigate the extraction solvent, the ratio of solid to liquid extraction time on the extraction rate of TTROG; tension sensor and BL-420 biological experiments multi-channel physiological signal acquisition and processing The inhibitory effect of different extracts on intestinal smooth muscle contraction in vitro was systematically recorded, and the extraction process of TTROG was evaluated by combining the biological activity with the extraction rate by using the weighted method. Results The optimal extraction conditions of TTROG were as follows: extraction solvent 80% ethanol, solid-liquid ratio 1:10, extraction time 2 h, extraction temperature 80 ℃, extraction times 3 times; Under optimal conditions, the extraction rate of TTROG was up to 42.12 mg / g; TTROG extract on the duodenum, jejunum, ileum, colon, four intestine have significant inhibition of intestinal smooth muscle contraction in a dose-dependent manner, the strongest effect on the jejunum, the inhibition rate up 41.96%. Conclusion The extraction rate of the optimized TTROG extraction process is high and stable, and the obtained TTROG extract can remarkably inhibit the contractile function of isolated rat smooth muscle in intestine.