痛泻要方对RIN-m细胞TRPA1表达及5-HT分泌的影响

Effects of Tongxie Yaofang on the expression of TRAP1 and secretion of 5-HT in RIN-m cells

目的:探讨痛泻要方对RIN-m细胞TRPA1表达及5-HT分泌的影响。方法:体外培养RIN-m细胞,对照组加入不含药物的RPMI-1640完全培养基,西药组加入浓度为0.5μg/m L钌红的RPMI-1640完全培养基,痛泻要方低(0.05%)、中(0.1%)、高(0.5%)浓度组分别加入相应浓度的含痛泻要方的RPMI-1640完全培养基,在静息与激活状态下,分别培养48h与72h,收集各组上清液及细胞,HPLC法检测5-HT浓度,Real-time PCR及Western blot法检测TRPA1表达。结果:在静息与激活状态下,与对照组比较,各组5-HT分泌量、TRPA1 mRNA与TRPA1蛋白表达均显著降低(P<0.05);与西药组比较,不同浓度痛泻要方各组5-HT分泌量、TRPA1 mRNA与TRPA1蛋白表达均显著升高(P<0.05);与本组培养48h比较,各组5-HT分泌量与TRPA1蛋白表达均显著升高(P<0.05);在一定时间内,TRPA1表达与5-HT分泌呈正相关(P<0.01)。结论:痛泻要方的作用机制可能与降低TRPA1介导的5-HT分泌有关。

Objective: To explore the effects of Tongxie Yaofang(TXYF) on the expression of TRAP1 and secretion of 5-HT in RIN-m cells. Methods: RIN-m cells were cultured in vitro and divided into five groups. The control group was cultured in RPMI-1640 medium, the Western medicine group was cultured in RPMI-1640 medium with 0.5μg/m L of ruthenium red, TXYF low concentration group(0.05%), TXYF middle concentration(0.1%) group and TXYF high concentration group(0.5%) were cultured in RPMI-1640 culture medium added with corresponding concentration of TXYF. After cultured for 48 hours and 72 hours, the supernatant and cells were collected, the concentration of 5-HT was detected by HPLC method and the expression of TRPA1 was detected by Real-time PCR and Western blot. Results: Compared with control group, 5HT secretion, TRPA1 mRNA and protein expression was decreased in each group during resting and activated condition(P<0.05). Western medicine group western medicine group, 5HT secretion, TRPA1 mRNA and protein expression was increased in TXYF each group(P<0.05). Compared with 48 hours culturing, 5HT secretion and TRPA1 protein expression was increased in each group(P<0.05). In a certain time, TRPA1 expression was positive correlation between 5HT secretion(P<0.01). Conclusion: The mechanism of TXYF may be associated with reducing TRPA1 expression and 5-HT secretion.