基于Wnt/β-catenin通路探讨当归补血汤防治化疗后骨髓造血细胞衰老的机制  

Exploration of Mechanism of Danggui Buxue Decoction in Preventing and Treating Bone Marrow Hematopoietic Cell Aging After Chemotherapy Based on Wnt/β-catenin pathway

作　　者：白莹 刘俊岑 王佳佳[1] 贾富霞[1] 王文娟[1] BAI Ying;LIU Juncen;WANG Jiajia;JIA Fuxia;WANG Wenjuan(School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学中医药学院,北京100069

出　　处：《中国中医基础医学杂志》2023年第11期1840-1846,共7页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然科学基金项目(81774176);国家重点研发计划“中医药现代化研究”专项(2018YFC1704102)。

摘　　要：目的 研究当归补血汤(Danggui Buxue decoction, DBD)对化疗后骨髓造血细胞衰老的改善作用与机制。方法 将60只雄性BALB/c小鼠随机分为空白对照组,模型对照组(100 mg/kg),阳性对照组(30μg/kg)和DBD高、中、低剂量组(24 g/kg、12 g/kg、6 g/kg),每组10只。阳性对照组皮下注射给予重组人粒细胞集落刺激因子(recombinant human granulocyte colony stimulating factor, rhG-CSF),DBD组按照体重灌胃,给药5 d后各组腹腔注射环磷酰胺(cyclophosphamide, CTX)3 d,建立化疗后骨髓细胞衰老模型,造模完成后,继续各类药物干预2 d。药物干预结束后检测小鼠外周血常规;称重器官;苏木素伊红(htoxylin eosin, HE)染色观察髂骨结构;ELISA测骨髓造血细胞SA-β-半乳糖苷酶(SA-β-galactosidase,SA-β-Gal)含量;Western blot法测定骨髓造血细胞中p21、p53、β-catenin、GSK-3β蛋白表达;实时荧光定量聚合酶链式反应(real time quantitative polymerase chain reaction,RT-qPCR)法测定骨髓造血细胞p21、p53、β-catenin、GSK-3β、c-myc m RNA表达。结果 与空白对照组相比较,模型对照组白细胞数、淋巴细胞数和中性粒细胞数下降(P<0.05);HE染色中骨髓中造血细胞数减少,造血组织间隙较大;骨髓造血细胞SA-β-Gal含量上升(P<0.01),p21、p53蛋白及m RNA表达上调(P<0.05);β-catenin蛋白及m RNA表达上调(P<0.01),c-myc m RNA表达上调(P<0.05)。与模型对照组相比较,DBD高剂量组白细胞数上升(P<0.05),SA-β-Gal含量下降(P<0.05),HE染色中骨髓中造血细胞数增多,造血组织间隙减小,p21、p53、β-catenin蛋白及m RNA表达均下调(P<0.05),GSK-3β m RNA表达上调(P<0.01);DBD中剂量组SA-β-Gal含量下降(P<0.01),DBD中剂量组β-catenin蛋白及m RNA表达下调(P<0.05);DBD低剂量组GSK-3βm RNA表达上调(P<0.05);β-catenin m RNA表达下调(P<0.01)。结论 DBD能保护化疗过程中的骨髓造血细胞并缓解其衰老,提示其机制与抑制Wnt/β-catenin通路有关。Objective To study the improvement effect and mechanism of Danggui Buxue decoction(DBD)on bone marrow hematopoietic cell senescence after chemotherapy.Methods Sixty male BALB/c mice were randomly divided into a blank control group,a model control group(100 mg/kg),a positive control group(30μg/kg),and DBD high,medium,and low dose groups(24 g/kg,12 g/kg,and 6 g/kg),with 10 mice in each group.The positive control group was given recombinant human granulocyte colony stimulating factor(rhG-CSF)by subcutaneous injection,and the DBD group was gavaged according to body weight,and after 5 d of administration,each group was injected intraperitoneally with cyclophosphamide(CTX)for 3 d to establish a positive control group.After 5 d of drug administration,each group was injected intraperitoneally with CTX for 3 d to establish a model of bone marrow cell senescence after chemotherapy,and after the modeling was completed,the intervention with various types of drugs was continued for 2 d.After the administration,the peripheral blood of the mice was detected,the organs were weighed.HE staining was used to observe the structure of the iliac bone.ELISA was used to measure the content of SA-β-galactosidase(SA-β-Gal)in bone marrow hematopoietic cells.Western blot was used to determine the protein expressions of p21,p53,β-catenin and GSK-3βin bone marrow hematopoietic cells and real time quantitative polymerase chain reaction(RT-qPCR)was used to determine the mRNA expressions of p21,p53,β-catenin,GSK-3βand c-myc in bone marrow hematopoietic cells.Results Compared with the blank control group,the number of white blood cells,lymphocytes and neutrophils in the model group decreased(P<0.05),the number of hematopoietic cells in the bone marrow decreased in HE staining and the space between hematopoietic tissues was larger.The content of SA-β-Gal in cells increased(P<0.01),the protein and mRNA expressions of p21,p53 andβ-catenin increased(P<0.05)and c-myc mRNA increased(P<0.01).Compared with the model control group,the number of w

关 键 词：当归补血汤 化疗 衰老 WNT/Β-CATENIN通路 p53/p21途径 

分 类 号：R285.5[医药卫生—中药学]