论文部分内容阅读
目的 探讨１８－α甘草酸二铵（甘利欣）的体外抗肝纤维化活性作用。方法 培养ＮＩＨ／３Ｔ３成纤维细胞，分别以含５％～２０％小牛血清的ＰＲＭＩ－１６４０培养液，或以无血清ＰＲＭＩ－１６４０培养液，或以含５％小牛血清ＰＲＭＩ－１６４０培养液稀释的１～４００μｇ／ｍ１甘利欣药物温育液，温育细胞。ＭＴＴ法检测药物对细胞的毒性作用与对小牛血清刺激的细胞增殖的影响， ＥＬＩＳＡ法测定细胞上清Ｉ型胶原含量，总蛋白测定（ＤＣ ｐｒｏｔｅｉｎ ａｓｓａｙ）试剂盒测定细胞层总蛋白含量，以校正细胞数。结果５０～４００μｇ／ｍ１浓度甘利欣抑制细胞ＭＴＴ转换，呈浓度依赖性增强，４００μｇ／ｍ１甘利欣呈现明显细胞毒作用。小牛血清一定浓度依赖性促进细胞增殖，而甘利欣对这种血清刺激细胞增殖作用明显呈浓度依赖性抑制。１０－２００μｇ／ｍ１甘利欣对Ｉ型胶原的分泌，亦有明显抑制作用。结论 甘利欣对ＮＩＨ／３Ｔ３成纤维细胞的增殖及Ｉ型胶原分泌有抑制作用，可能具有抗肝纤维化活性。
Objective To investigate the anti-hepatic fibrosis activity of 18-α-diammonium glycyrrhizinate (Glycyrrhizin) in vitro. Methods NIH / 3T3 fibroblasts were cultured in RPMI-1640 medium containing 5% ~ 20% fetal bovine serum or PRM-1640 serum-free medium or PRMI-1640 medium containing 5% fetal bovine serum Dilute 1 ~ 400μg / m1 Glycyrrhizin incubation solution, incubate the cells. MTT assay was used to determine the cytotoxicity of the drug on the cell proliferation induced by bovine serum. The content of type I collagen in the cell supernatant was measured by ELISA. The total protein content in the cell layer was determined by the kit of DC protein assay Calibrate cell number. Results The concentrations of 50 ~ 400μg / ml Glycyrrhizin inhibited the MTT conversion in a concentration-dependent manner, and 400μg / ml Glycyrrhizin showed obvious cytotoxicity. Bovine calf serum concentration-dependently promote cell proliferation, and salivariamine on this serum-stimulated cell proliferation in a significant concentration-dependent inhibition. 10-200μg / m1 Glycyrrhizin on the secretion of type I collagen, also significantly inhibited. Conclusion Glycyrrhizin can inhibit the proliferation and type I collagen secretion of NIH / 3T3 fibroblasts and may have anti-hepatic fibrosis activity.