摘要
采用AB-8大孔树脂纯化沙棘黄酮初提物,用10%、30%、50%乙醇溶液梯度洗脱,以总黄酮含量和黄酮苷元含量为指标,筛选纯化样品;采用MTT法、实时无标记细胞分析技术检测沙棘黄酮样品体外对人前列腺癌PC-3细胞的增殖抑制作用,流式细胞技术检测细胞凋亡和细胞周期情况,Western blot检测Bax和Bcl-2蛋白表达情况。实验结果表明50%乙醇梯度洗脱样品(S50)中,总黄酮含量达到25.42%;水解后得到SH50样品,其槲皮素、异鼠李素含量分别达到5.03%、18.64%;25μg/m L的SH50样品对PC-3细胞即有增殖抑制作用,且呈剂量和时间依赖性;同时25μg/m L的SH50样品作用48 h即可显著诱导细胞凋亡(P<0.01),并将细胞周期阻滞在G2/M期,此外,样品可提高Bax蛋白和降低Bcl-2蛋白的表达。说明沙棘黄酮SH50样品对体外人前列腺癌PC-3细胞具有抑制增殖并诱导细胞凋亡作用,其机制可能与阻滞细胞周期,调节Bax和Bcl-2蛋白的表达有关。
Using AB-8 macroporous resin as carrier and gradiently eluted by 10%,30%,50%ethanol,the purified samples were screened with the content of total flavonoids and flavonoid aglycones.The MTT assay and RTCA were used to examine inhibitory effect of flavonoids from Sea buckthorn on PC-3 cells.Flow-cytometry was used to analyze the cell apoptosis and cell cycle of the PC-3 cells.The expressions level of Bax and Bcl-2 were detected by Western blot.The results showed that in the gradient elution of 50%ethanol solution(S50),the content of total flavonoids was 25.42%,hydrolyzing it to get SH50 sample,which quercetin and isorhamnetin content reached 5.03%and 18.64%respectively;SH50(25μg/mL)can inhibit the activity of PC-3 cells and showed the feature of time/dose dependent;25μg/mL SH50 can also induce the apoptosis of PC-3 cells significantly(P<0.01)and block the cell cycle at G2/M stage after treated 48h.Moreover,SH50 can increase the activities of Bax but decrease the expression level of Bcl-2.These resuls demonstrated that SH50 can inhibit the proliferation and induce cell apoptosis of PC-3 cells in vitro,the mechanism might be related with the blocking of the cell cycle and regulating the expression levels of Bax and Bcl-2.
作者
赵波
向晓玲
王微
李春松
吴晓琴
沈建福
ZHAO Bo;XIANG Xiao-lin;WANG Wei;LI Chun-song;WU Xiao-qin;SHEN Jian-fu(Colleage of Biosystems Engineering and Food Science,Zhejiang University,Hangzhou 310058,China;Institute of food and Drug Administration of Jinhua,Jinhua 321000,China)
出处
《天然产物研究与开发》
CAS
CSCD
北大核心
2018年第1期27-32,160,共7页
Natural Product Research and Development
基金
国家自然科学基金面上项目(J20120653)
关键词
沙棘黄酮
人前列腺癌
PC-3细胞
抑制增殖
sea buckthorn flavonoids
human prostate cancer
PC-3 cells
inhibit the proliferation
