摘要
目的:探讨芪苓白头翁汤对弥漫大B细胞淋巴瘤(DLBCL)细胞增殖、凋亡、非受体型酪氨酸蛋白激酶2/信号转导及转录激活因子3(JAK2/STAT3)信号通路及炎症因子白细胞介素^(-1)0(IL^(-1)0)的影响。方法:以人DLBCL细胞OCI-LY10、U2932细胞为研究对象,细胞增殖与活性检测法(CCK-8)检测细胞增殖情况,并计算出0、4.6、9.3、18.7、37.5、75、150 mg·L^(-1)芪苓白头翁汤处理OCI-LY10、U2932细胞24 h后的半抑制浓度(IC_(50))分别为9.33、16.13 mg·L^(-1)。后续相关实验根据芪苓白头翁汤作用于OCI-LY10、U2932细胞24 h半抑制浓度(IC_(50)),选用芪苓白头翁汤9.5、19、38 mg·L^(-1)开展实验。用胱天蛋白酶(Caspase)-3、Caspase-8、Caspase-9酶原活化检测试剂盒检测经0、9.5、19、38 mg·L^(-1)芪苓白头翁汤处理OCI-LY10、U2932细胞24 h后OCI-LY10、U2932细胞中Caspase-3、Caspase-8、Caspase-9的酶原活化情况。酶联免疫吸附测定法(ELISA)检测经0、9.5、19、38 mg·L^(-1)芪苓白头翁汤处理OCI-LY10、U2932细胞24 h后OCI-LY10、U2932细胞中IL^(-1)0炎症因子表达情况。流式细胞仪检测不同浓度芪苓白头翁汤作用OCI-LY10、U2932细胞24 h后细胞凋亡率及细胞周期。蛋白免疫印迹法(Western blot)检测0、9.5、19、38 mg·L^(-1)芪苓白头翁汤作用OCI-LY10、U2932细胞24 h,OCI-LY10、U2932细胞中B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、活化的聚腺苷二磷酸核糖聚合酶(cleaved PARP)、活化的Caspase-3(cleaved Caspase-3)凋亡蛋白表达情况及JAK2、STAT3、磷酸化(p)-JAK2、p-STAT3通路蛋白表达情况,不同浓度药物作用OCI-LY10、U2932细胞24 h,OCI-LY10、U2932细胞中癌基因(c-Myc)蛋白表达情况。结果:芪苓白头翁汤作用于OCI-LY10、U2932细胞24 h,与空白组比较,芪苓白头翁汤各组细胞增殖均受到明显抑制(P<0.05,P<0.01);9.5、19、38 mg·L^(-1)芪苓白头翁汤组Caspase-3、Caspase-8、Caspase-9酶原显著活化(P<0.01),细胞凋亡增加(P<0.05,P
Objective:To investigate the effect of Qiling Baitouweng Tang(QLBTWT)on proliferation and apoptosis,Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway and interleukin^(-1)0(IL^(-1)0)in diffuse large B-cell lymphoma(DLBCL).Method:With human DLBCL cells OCI-LY10 and U2932 as research objects,cell proliferation was detected by cell counting kit-8(CCK-8)assay.After treatment with 0,4.6,9.3,18.7,37.5,75,150 mg·L^(-1)QLBTWT for 24 h,the halfinhibitory concentration(IC_(50))of OCL-LY10 and U2932 cells was calculated to be 9.33,16.13 mg·L^(-1),respectively,based on which,9.5,19,38 mg·L^(-1)QLBTWT were selected for subsequent experiments.After 0,9.5,19,38 mg·L^(-1)QLBTWT treatment for 24 h,the zymogen activities of Caspase-3,Caspase-8 and Caspase-9 in OCI-LY10 and U2932 cells were detected using corresponding activity assay kits(colorimetric),and the IL^(-1)0 expression was detected by enzyme-linked immuno sorbent assay(ELISA).The apoptosis rate and cell cycle of OCI-LY10 and U2932 cells treated with different concentrations of QLBTWT for 24 h were detected by flow cytometry.The expressions of apoptosis-related proteins[B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),cleaved poly adenosine diphosphate ribose polymerase(cleaved PARP),cleaved Caspase-3],JAK2,STAT3,phospho-JAK2(p-JAK2),phospho-STAT3(p-STAT3)pathway proteins,and c-Myc protein in OCL-LY10 and U2932 cells after 24 h treatment with 0,9.5,19,38 mg·L^(-1)QLBTWT were all tested by Western blot.Result:After QLBTWT treatment on OCI-LY10 and U2932 cells for 24 h,cell proliferation was inhibited in each QLBTWT group compared with that in the control group(P<0.05,P<0.01).The zymogens of Caspase-3,Caspase-8 and Caspase-9 were activated(P<0.01),and there was an increase in cell apoptosis(P<0.05,P<0.01)and cell cycle arrest at Gap phase1(G1)phase in 9.5,19 and 38 mg·L^(-1)QLBTWT group(P<0.05,P<0.01).After 9.5,19 and 38 mg·L^(-1)QLBTWT treatment on OCI-LY10 and U2932 cells for 24 h,the expressions of Bcl-2,p-JAK2
作者
刘宁
战昕卓
于慧
陈晓丽
孔祥图
倪海雯
LIU Ning;ZHAN Xinzhuo;YU Hui;CHEN Xiaoli;KONG Xiangtu;NI Haiwen(The First School of Clinical Medicine of Nanjing University of Chinese Medicine,Nanjing 210023,China;Affiliated Hospital of Nanjing University of Chinese Medicine,Jiangsu Province Hospital of Chinese Medicine,Nanjing 210029,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第13期10-19,共10页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家中医药管理局第四批全国中医优秀人才研修项目(国中医药办人教函〔2017〕24号)
江苏省中医药管理局课题(YB2017014)
江苏省卫生健康委重点项目(ZD2021040)
江苏省研究生科研与实践创新计划项目(KYCX22_1891)。
