表儿茶素及其衍生物表儿茶素没食子酸酯通过MAPK-ERK44/42通路抑制前列腺间质细胞增殖

Epicatechin and its derivate Epicatechin gallate inhibit prostatic stromal cell proliferation through selective MAPK-ERK44/42 pathway

关键词:  表儿茶素没食子酸酯  表儿茶素  良性前列腺增生  细胞增殖  MAPK通路

Key words:  Epicatechin gallate  Epicatechin  benign prostatic hyperplasia  cell proliferation  MAPK pathway

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10 μmol/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。

Abstract:

[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 C6-Ceramide (C6) or dual agonist Anisomycin (ANI) of P38, SAPK/JNK.[Results] Both ECG and EC (1 nmol/L to 10 μmol/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 altered. After the adding of C6 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 stromal cell proliferation through ERK44/42 pathway, which provides evidence for their mechanism on benign prostatic hyperplasia prevention.