迷迭香酸抑制人多发性骨髓瘤ARH-77细胞增殖的作用机制  

Mechanism of rosmarinic acid inhibiting proliferation of human multiple myeloma ARH-77 cells

作　　者：娄诤 赵肖涯 杜丽君[2] 王大维[3] LOU Zheng;ZHAO Xiao-ya;DU Li-jun;WANG Da-wei(The Second Clinical Medical School;School of Life Sciences;Basic Medical School,Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学第二临床医学院,浙江杭州310053 [2]浙江中医药大学生命科学学院,浙江杭州310053 [3]浙江中医药大学基础医学院,浙江杭州310053

出　　处：《中国药理学通报》2018年第8期1078-1082,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金面上项目(No 81373633);浙江省自然科学基金资助项目(No LY18H270014)

摘　　要：目的研究迷迭香酸(rosmarinic acid,RosA)对ARH-77细胞的抑制作用及其机制。方法采用CCK-8法检测RosA对ARH-77细胞增殖的影响,并由该结果确定实验组用药浓度分别为:20、40、80μmol·L^(-1),作用时间48 h;通过流式细胞术检测凋亡率;采用qPCR法检测Fas、caspase-8、caspase-9 mRNA表达;采用分光光度法检测caspase-8的活性;Western blot法检测t Bid、Fas、活性caspase-3和活性caspase-9的蛋白水平。结果 RosA能明显抑制ARH-77细胞的增殖;与对照组相比:各实验组均主要发生早期凋亡(P<0.01);各实验组活性的caspase-3、caspase-9水平均明显升高(P<0.01),caspase-8 mRNA表达也明显上调,但其蛋白活性仅在Ros A 80μmol·L^(-1)时明显升高(P<0.01);在Ros A(40、80μmol·L^(-1))组,caspase-9 mRNA表达明显上调(P<0.05,P<0.01),t Bid的蛋白表达也明显升高(P<0.01);Fas mRNA和蛋白表达仅在Ros A 80μmol·L^(-1)组明显升高(P<0.01)。结论 RosA诱导ARH-77细胞早期凋亡而导致了增殖抑制,死亡受体途径和线粒体途径可能共同参与了Ros A诱导的ARH-77细胞凋亡。Aim To study the inhibitory effect of rosmarinic acid（RosA） on ARH-77 cells and its mechanism. Methods The effect of RosA on the proliferation of ARH-77 cells was detected by CCK-8 method.The results confirmed that the concentration of the drug in the experimental group was 20,40 and 80 μmol ·L-1,respectively,for 48 h; the apoptosis rate was detected by flow cytometry; mRNA expression of Fas,caspase-8,and caspase-9 was measured by qPCR; the activity of caspase-8 was measured by spectrophotometry; t Bid,Fas,active caspase-3,and active caspase-9 were detected by Western blot. Results RosA obviously inhibited the proliferation of ARH-77 cells. Compared with control group,early apoptosis was observed in all experimental groups（P〈0. 01）. In each experimental group,the levels of active caspase-3 and caspase-9 markedly increased（P〈0. 01）,caspase-8 mRNA expression was also significantly up-regulated,but its protein activity only evidently increased at 80μmol· L-1（P〈0. 01）; at 40 and 80 μmol · L-1 groups,the mRNA expression of caspase-9 was significantly up-regulated（P〈0. 05,P〈0. 01）,and the protein expression of t Bid also notably increased（P〈0. 01）; the mRNA and protein expression of Fas dramatically increased only at 80 μmol·L-1 group（P〈0. 01）. Conclusions The early apoptosis of ARH-77 cells induced by RosA leads to the inhibition of proliferation. The death receptor and mitochondrial pathways may participate in the apoptosis of ARH-77 cells induced by RosA.

关 键 词：迷迭香酸 ARH-77细胞 TBID FAS CASPASE-3 CASPASE-9 CASPASE-8 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学]