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目的探讨四君子汤(SJZT)对多胺合成特异性抑制剂二氟甲基鸟氨酸(α-difluoromethyornithine,DFMO)作用下的Caco-2人结直肠腺癌细胞微绒毛及腐胺(PUT)、精眯(SPD)、精胺(SPM)等多胺含量的影响,阐明SJZT的肠黏膜修复机制。方法大鼠灌服SJZT 17 g·kg~(-1),空白对照组给予等量生理盐水,每日2次,连续3 d,末次给药1 h后腹主动脉取血,制备SJZT含药血清。以Caco-2细胞为研究对象,实验分为对照组、DFMO组、SPD+DFMO组、SJZT(10%,5%,2.5%含药血清)+DFMO组。采用MTT法检测细胞增殖;透射电镜观察细胞微绒毛结构;柱前衍生-高效液相色谱(HPLC)法检测细胞内多胺含量。结果 DFMO组与对照组比较,DFMO对Caco-2细胞增殖、微绒毛的生长及多胺含量均有明显抑制作用,差异有统计学意义(P<0.05,P<0.01);而SPD及SJZT各剂量组能明显促进Caco-2细胞生长,改善微绒毛的生长抑制,提高细胞内多胺的含量。结论SJZT对肠黏膜损伤的修复作用可能与提高细胞多胺含量,促进Caco-2细胞增殖,改善微绒毛结构有关。
Objective To investigate the effects of SJZT on the microvilli and putrescine (PUT) of Caco-2 human colorectal adenocarcinoma cells under the action of DFMO, a polyamine specific inhibitor. Fine squint (SPD), spermine (SPM) and other polyamine content, clarify the SJZT intestinal mucosal repair mechanism. Methods SJZT 17 g · kg -1 was administered to rats in the control group. The rats in the blank control group were given normal saline twice a day for 3 consecutive days. Blood was collected from the abdominal aorta 1 h after the last administration, serum. The Caco-2 cells were divided into control group, DFMO group, SPD + DFMO group and SJZT (10%, 5%, 2.5% serum containing) + DFMO group. Cell proliferation was detected by MTT assay. Microvilli structure was observed by transmission electron microscopy. Pre-column derivatization-high performance liquid chromatography (HPLC) was used to detect intracellular polyamines content. Results Compared with the control group, DFMO had a significant inhibitory effect on proliferation, microvilli growth and polyamines content in Caco-2 cells (P <0.05, P <0.01) The dose group can significantly promote the growth of Caco-2 cells, improve the growth inhibition of microvilli and increase the intracellular polyamines content. Conclusion The repair effect of SJZT on intestinal mucosal injury may be related to increasing the content of polyamines, promoting the proliferation of Caco-2 cells and improving the structure of microvilli.