摘要
[目的]探究锁阳中表儿茶素(EC)及其衍生物表儿茶素没食子酸酯(ECG)对前列腺间质细胞WPMY-1增殖的影响及其机制。[方法]体外培养WPMY-1细胞,加入不同剂量的ECG和EC,用噻唑蓝(MTT)法检测对细胞增殖的影响,用聚合酶链反应(PCR)和蛋白免疫印迹(Western blotting)法检测对增殖相关抗原PCNA mRNA和蛋白水平表达的影响;用Western blotting法分别检测ECG和EC对MAPK通路ERK44/42、P38、SAPK/JNK激活的抑制作用;加入ERK44/42激动剂C6-Ceramide(C6)及P38、SAPK/JNK联合激动剂Anisomycin(ANI)后,用MTT法检测对ECG和EC抑制细胞增殖的影响。[结果]1 nmol/L至10μmol/L的ECG和EC均可以明显抑制WPMY-1细胞的增殖(P<0.01)。10 nmol/L的ECG和EC可以明显抑制WPMY-1细胞中PCNA mRNA和蛋白水平的表达(P<0.05)。ECG和EC均能抑制ERK44/42和SAPK/JNK的磷酸化,但对P38的磷酸化作用不明显;加入C6后,ECG和EC抑制WPMY-1增殖的能力明显降低(P<0.05或P<0.01),加入ANI后,则细胞增殖能力改变不明显。[结论]ECG和EC通过ERK44/42抑制前列腺间质细胞增殖,这可能是其抗良性前列腺增生的重要机制之一。
[Objective] To study the effect and mechanism of Epicatechin （EC） and its derivate Epicatechin gallate （ECG） in cynomorium songaricum on the proliferation of prostrate stromal cell line WPMY-1. [Methods] WPMY-1 cells were treated with different doses of ECG and EC in vitro. MTT assay was performed to detect the effects of ECG and EC on WPMY-1 cell proliferation. PCR and western blot were performed to detect the mRNA and protein expressions of proliferating cell nuclear antigen proliferating cell nuclear antigen（PCNA） separately in WPMY-1 cells treated with ECG or EC. The phosphorylation levels of MAPK signaling pathway ERK44/42, P38, and SAPK/ JNK were then detected by western blot in WPMY-1 cells after treatment with ECG or EC. The effects of different MAPK signalings on ECG and EC-suppressed cell proliferation were investigated in WPMY-1 cells after treatment with or without ERK44/42 agonist C 6- C e - ramide （C 6 ） or dual agonist Anisomycin （ANI） of P38, SAPK/JNK. [Results] Both ECG and EC （ 1 nmol / L to 1 0 pmol / L ） inhibited the proliferation of WPMY-1 cells significantly （P〈0.01） and PCNA expressions in mRNA and protein levels in WPMY-1 cell（P〈0.05）. the constitutive phosphorylation levels of ERK44/42 and SAPK/JNK were significantly suppressed by ECG and EC, which of P38 was not al-tered. After the adding of C 6 blocked ECG and EC-induced suppression of WPMY-1 cell proliferation （P〈0.05 or P〈0.01）. The change of cell proliferation was not significant after the adding of ANI. [Conclusion] ECG and EC inhibit prostatic s t r omal cell proliferation through ERK44/42 pathway, which provides evidence for their mechanism on benign prostatic hyperplasia prevention.
出处
《天津中医药》
CAS
2017年第3期200-203,共4页
Tianjin Journal of Traditional Chinese Medicine
基金
国家自然科学基金青年科学基金项目(81303293)
天津市应用基础及前沿技术研究计划青年基金项目(14JCQNJC13400)
