基于wnt1信号通路探讨黑枸杞对肝干细胞的影响  

Discussion on the Effect of Lycium Ruthenicum on Hepatic Stem Cells Based on Wnt1 Signaling Pathway

作　　者：马力 马静 张常喜 MA Li;MA Jing;ZHANG Changxi(Ningxia Hui Autonomous Region Hospital of Traditional Chinese Medicine,Research Institute of Traditional Chinese Medicine,Ningxia,Yinchuan 750021,China)

机构地区：[1]宁夏回族自治区中医医院暨中医研究院,宁夏银川750021

出　　处：《光明中医》2022年第22期4066-4070,共5页GUANGMING JOURNAL OF CHINESE MEDICINE

基　　金：宁夏自然科学基金项目(No.NZ17214)。

摘　　要：目的基于相关信号通路探讨黑枸杞对肝干细胞增殖分化的影响。方法先制备药物血清,并将肝干细胞系WB-F344分为3组:空白组加10%磷酸盐缓冲液(PBS)、空白血清处理组加不含药胎牛血清(FBS)10%、实验药物血清处理组加入客户提供的含药血清处理(含药血清量10%),激光共聚焦观察肝干胞标记物甲胎蛋白(AFP)、肝细胞标记物白蛋白(ALB)、以及胆管上皮细胞标记物角蛋白(CK-19)以观察肝干细胞诱导分化的变化,药物诱导48 h后,过碘酸雪芙染色,QPCR和WB检测CYP1A、Wnt1、Notch2的表达。结果药物诱导48 h后,激光共聚焦结果显示肝干胞标记物AFP、ALB、以及CK-19均有表达。糖原PAS染色结果可知与空白对照组相比,不加药血清组阳性物质的表达少于空白对照组,加药血清组阳性物质少于空白对照组。Q-PCR检测结果表明:相较于空白对照组,不加药血清组和加药血清组中Wnt1、CYP1A1、Notch2的表达均高于空白对照组;与不加药血清组相比,加药血清组CYP1A1和Wnt1的表达均低于不加药血清组。WB检测结果表明与空白对照组相比不加药血清组和加药血清组CYP1A1、Notch2和Wnt1的表达均高于空白对照组;与不加药血清组相比,加药血清组CYP1A1、Notch2和Wnt1的表达均高于不加药血清组。结论黑枸杞可以通过抑制Wnt1信号表达从而对肝脏起到保护作用。Objective To explore the effect of lycium ruthenicum on the proliferation and differentiation of liver stem cells based on related signal pathways.Methods Preparing drug serum firstly,and the liver stem cell line WB-F344 were divided into blank group adding 10%PBS,blank serum treatment group adding drug-free FBS10%and experimental serum serum treatment group.Adding the drug-containing serum treatment provided by the customer(the amount of drug-containing serum 10%),laser confocal microscopy was used to observe liver stem cell marker alpha-fetoprotein AFP,hepatocyte marker albumin ALB,bile duct epithelial cell marker keratin CK-19 observation.The changes of hepatic stem cells induced differentiation,48 hours after drug induction,periodic acid Sheffield staining,QPCR and WB were used to detect the expression of CYP1A,Wnt1,Notch2.Results 48 hours after drug induction,the laser confocal results showed that liver stem cell marker alpha-fetoprotein AFP,hepatocyte marker albumin ALB,and bile duct epithelial cell marker keratin CK-19 were all expressed.The results of glycogen PAS staining showed that compared with the blank control group,the expression of positive substances in the non-medicated serum group was less than that of the blank control group,and the positive substances in the medicated serum group were less than the blank control group.The Q-PCR test results showed that compared with the blank control group,the expressions of Wnt1,CYP1A1,and Notch2 in the non-medicated serum group and the medicated serum group were higher than those in the blank control group;compared with the non-medicated serum group,the medicated serum group.The expressions of CYP1A1 and Wnt1 were lower than those of the serum group without medicine.The WB test results showed that the expressions of CYP1A1,Notch2 and Wnt1 in the serum group without medicine and the serum group were higher than those in the control group compared with the blank control group.Compared with the serum group without medicine,the expression of CYP1A1,Notch2 in th

关 键 词：黑枸杞 肝干细胞系 WNT1 CYP1A1 NOTCH2 实验研究 

分 类 号：R285[医药卫生—中药学]