沙芭特在非细菌性前列腺炎症中的抗炎、抗纤维化及促进上皮细胞凋亡作用的研究

摘要

Objective To explore the role of saw palmetto extract on non-bacterial prostatitis. Methods Prostate epithelial cell RWPE-1 was cultured with or without conditioned medium from M1 type THP-1 cell and treated with saw palmetto extract.Cell viability was measured by MTS and cell apoptosis was assessed with flow cytometry and the expression of inflammation,fibrosis and ap-optosis-related molecules (IL-1β,IL-1 7,TNF-α,Fas,COX2,Bax,TGF-β,DDR1,α-SMA and CollagenⅠ)were detected by RT-PCR,ELISA and Western blot.In the Xiaozhiling-induced non-bacterial prostatitis rat model,mechanical paw withdrawal thresholds were tested with von Frey fila-ments before and after saw palmetto extract treatment.At last,blood from vena cava was collected under anesthesia,then the prostate tissues were extracted,weighed and further investigated with masson staining,ELISA and IHC. Results Saw palmetto extract inhibited cell proliferation and induced the apoptosis of RWPE-1,M1 type THP-1 cell conditioned medium upregulated the gene ex-pression of IL-1β,TNF-αand COX-2 which could be partly impaired by saw palmetto extract treat-ment.Besides,saw palmetto extract treatment inhibited the expression of COX-2,DDR1,SMA, CollagenⅠand elevated the expression of Fas and Cleaved-caspase3.In the Xiaozhiling-induced non-bacterial prostatitis rat model,saw palmetto extract group showed higher mechanical paw withdraw-al thresholds,lower prostate index,and lower expression of TGF-β,TNF-α,COX-2 and CD68 in prostate tissues. Conclusions Saw palmetto extract could inhibit inflammation,fibrosis and cell growth of prostate cells under the circumstance of inflammation both in vitro and in vivo.%目的 探讨锯叶棕果实提取物沙芭特在非细菌性前列腺炎中的效应及其机制.方法 前列腺上皮细胞RWPE-1经M1表型的单核巨噬细胞THP-1细胞上清液共培养,治疗组加用沙芭特(10μg/ml)培养,MTS检测细胞增殖,流式细胞术检测细胞凋亡,RT-PCR、ELISA及West-ern blot检测炎症、纤维化及凋亡相关的蛋白分子(IL-1β、IL-17、TNF-α、Fas、COX2、Bax、TGF-β、DDR1、α-SMA及CollagenⅠ)的表达.消痔灵前列腺注射诱导大鼠非细菌性前列腺炎模型,用von Frey纤维检测沙芭特治疗前后各组大鼠的痛觉敏感性,然后麻醉SD大鼠,腔静脉取血后摘取前列腺组织并称重,免疫组化及ELISA检测炎症因子的表达,masson染色检测纤维化.结果 沙芭特抑制RWPE-1的增殖并促进其凋亡.M1表型THP-1条件培养液能促进上皮细胞表达IL-1β、TNF-α及COX-2,沙芭特处理能抑制炎症刺激所致的炎症因子表达,Western blot检测表明沙芭特处理能抑制COX-2、DDR1、SMA及CollagenⅠ的表达,增加Fas及Cleaved-caspase 3的表达.消痔灵100μl每侧叶注射可以成功诱导前列腺炎症模型,沙芭特60mg/kg能减少大鼠前列腺组织炎症因子的表达,缩小前列腺体积,降低大鼠痛觉敏感性.结论 沙芭特能抑制前列腺上皮细胞RWPE-1的增殖并促进其凋亡,减小前列腺体积,抑制炎症因子及纤维化相关分子的表达,减轻前列腺炎症及纤维化从而起到治疗作用.