针灸对CTX荷瘤小鼠骨髓细胞中Notch信号通路差异基因jag1、notch2的影响  

Effects of Acupuncture and Moxibustion on Regulation of Jag1 and Notch2 in Notch Signaling Pathway of Bone Marrow Cells in CTX Chemotherapy Tumor Mice

作　　者：路玫 牛云云[2] 曹大明[3] 于冬冬[2] 滕迎春[3] LU Mei;NIU Yunyun;CAO Daming;YU Dongdong;TENG Yingchun(School of International Education,Henan University of TCM,Zhengzhou 450046,Henan,China;School of Acupuncture andMoxibustion,Henan University of TCM,Zhengzhou 450046,Henan,China;The First Affiliated Hospital of Henan University of TCM,Zhengzhou 450008,Henan,China)

机构地区：[1]河南中医药大学国际教育学院,河南郑州450046 [2]河南中医药大学针灸推拿学院,河南郑州450046 [3]河南中医药大学第一附属医院,河南郑州450008

出　　处：《中华中医药学刊》2020年第1期1-5,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81674080)。

摘　　要：目的采用荷瘤小鼠为研究对象,以Notch信号通路为切入点,通过观察针刺与艾灸干预后环磷酰胺(CTX)模型荷瘤小鼠骨髓细胞中Notch信号通路差异基因jag1、notch2的表达、蛋白量以及mRNA转录情况,探讨针灸改善荷瘤小鼠骨髓抑制的相关作用机制及途径,为临床中针灸减轻肿瘤化疗患者骨髓抑制、提高生存质量及改善造血机能方面提供实验室依据及治疗思路。方法将40只昆明种雄性、清洁级小鼠自由饲养3 d后,在温度20~25℃,湿度60%左右的无菌条件下进行移植瘤模型制备,植瘤7 d后,对40只带瘤小鼠进行尾静脉采血,查其白细胞(WBC)总数并选取WBC数目接近正常范围,瘤体生长良好的荷瘤小鼠32只。将选取的荷瘤小鼠,随机分4组,每组8只,即空白组(A),模型组(B),针疗组(C),灸疗组(D)。B、C、D组均一次性腹腔注射环磷酰胺(CTX)150 mg/kg,A组给予相同体积的生理盐水。C、D组选取"大椎""膈俞""肾俞""足三里"穴位分别施以针刺、艾灸,A、B组每日陪同固定,不治疗。在课题组前期初步筛选的相关差异基因jag1、notch2的基础上,进一步运用免疫组化法、Real-time PCR法、Western Blot法全面深入地测定荷瘤小鼠骨髓细胞中Notch信号通路jag1、notch2基因的变化。结果(1)与空白组比,模型组中jag1和notch2基因蛋白表达升高,具有统计学意义(P<0.05);与模型组比,针疗组与灸疗组中jag1和notch2基因蛋白表达下降,具有统计学意义(P<0.05);(2)与空白组比,模型组中jag1、notch2基因mRNA表达升高;与模型组比,针疗组与灸疗组中jag1、notch2基因mRNA表达降低,但均无统计学意义;(3)与空白组比,模型组中jag1、notch2基因蛋白含量升高且具有统计学意义(P<0.05);与模型组比,针疗组、灸疗组jag1、notch2基因蛋白含量均降低,且具有统计学意义(P<0.05)。结论对Notch信号通路进行靶向调控,下调荷瘤小鼠骨髓细胞Notch信号通路中关键基因jag1、notcObjective To explore the related effect of acupuncture and moxibustion improving the mechanism of bonemarrow inhibition of tumor-bearing mice so as to provide laboratory basis and treatmentfor the clinical acupuncture reducing bone marrow suppression,improving the quality of survivaland the hematopoietic function by adopting tumor-bearing mice as the objects and Notch signaling pathway as the breakthrough point and the observation of gene jag1 difference of Notch signaling pathway,the expression of notch2,amount of protein and mRNA transcription situation after acupuncture and moxibustion.Methods Forty Kunming malemice at the clean level were free fed for 3 d.And then the model was prepared in the aseptic environment with temperature 20~25℃and humidity 60%.Seven days after implanting the tumor,the tail vein blood was taken out to check the total number of white blood cells(WBC),and we selected the WBC number which was close to the normal range.There were only 32 tumor-bearing mice with tumors growing well.These tumor-bearing mice were randomly divided into 4 groups:blank group(A),model group(B),acupuncture group(C)and moxibustion group(D),8 in each.In group B,C and D,cyclophosphamide(CTX)150 mg/kg was injected into the abdominal cavity once,and the same volume of saline was given to group A.In group C and D,acupuncture and moxibustion were respectively applied at acupoints of"Duzhui"(GV14),"Geshu"(BL17),"Shenshu"(BL23)and"Zusanli"(ST36).Group A and B were accompanied daily for fixation without treatment.On the basis of the related differential genes jag1 and notch2 that were preliminarily screened by the research team,immunohistochemistry,real-time PCR and Western Blot were further used to comprehensively and deeply determine the changes of the Notch signaling pathways jag1 and notch2 in the bone marrow cells of mice with tumor.Results(1)Compared with group A,the protein expressions of jag1 and notch2 genes in group B were increasedwith statistical significance(P<0.05).Compared with group B,the protein expressi

关 键 词：针灸 荷瘤小鼠 NOTCH信号通路 jag1 NOTCH2 

分 类 号：R245.3[医药卫生—针灸推拿学]