摘要
目的探讨黄连素对Lewis肺癌(Lewis lung cancer,LLC)细胞小鼠移植瘤的放射增敏效应。方法建立肺癌LLC细胞小鼠移植瘤模型;移植瘤平均体积增至约100 mm^(3)时(第0天),将24只小鼠随机分为4组:对照组、黄连素组(5 mg/kg黄连素)、照射组(8 Gy)、黄连素+照射组(5 mg/kg黄连素+8 Gy)。第2天开始,黄连素组和黄连素+照射组每隔1 d腹腔注射黄连素5 mg/kg,对照组和照射组腹腔注射等体积的生理盐水,给药第6次后(第12天),照射组及黄连素+照射组予局部单次8 Gy照射。第13天起各组均不予处理,每隔2 d测量1次小鼠肿瘤体积。待对照组小鼠肿瘤体积增至2000 mm^(3)时(第24天),处死小鼠,测量移植瘤体积,计算肿瘤体积抑制率;并对移植瘤标本进行免疫荧光染色中性粒细胞胞外诱捕网(neutrophil extracellular traps,NETs)相关蛋白瓜氨酸组蛋白3(citrullinated histone H3,CitH3)、髓过氧化物酶(myeloperoxidase,MPO)。第24天取小鼠眼内眦静脉血,离心取血清,用酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测小鼠肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、γ干扰素(interferon-γ,IFN-γ)表达水平。结果与对照组相比,黄连素组、照射组及黄连素+照射组小鼠移植瘤体积减小(P<0.01),黄连素组共表达CitH3、MPO细胞荧光强度降低(P<0.01)、TNF-α、IFN-γ表达水平降低(P<0.01),照射组及黄连素+照射组共表达CitH3、MPO细胞荧光强度增加(P<0.01)、TNF-α、IFN-γ表达水平升高(P<0.01)。与黄连素组比较,黄连素+照射组小鼠肿瘤体积减小(P<0.01),共表达CitH3、MPO细胞荧光强度增加(P<0.01)、TNF-α、INF-γ表达水平增加(P<0.01);与照射组比较,黄连素+照射组小鼠肿瘤体积减小(P<0.01),共表达CitH3、MPO细胞荧光强度减少,TNF-α、INF-γ表达水平降低(P<0.01)。结论黄连素对肺癌LLC细胞小鼠移植瘤具有放射增敏作用,可能与其抑制NETs的形成有关。
Objective To investigate the radiosensitising effect of safranin on transplanted tumors in mice with Lewis lung cancer(LLC) cells.Methods A transplanted tumor model was established in mice with Lewis lung cancer(LLC) cells.When the average volume of the transplanted tumor was increased to about 100 mm^(3)(which was defined as day 0),24 mice were randomly divided into four groups:control group,berberine group(5 mg/kg berberine),irradiation group(8 Gy),and berberine+irradiation group(5 mg/kg berberine+8 Gy).From day 2,the mice in berberine group and berberine+irradiation group were intraperitoneally injected with 5 mg/kg berberine every other day,and the mice in control group and irradiation group were intraperitoneally injected with an equal volume of saline,and the mice in irradiation group and berberine+irradiation group were irradiated with 8 Gy locally at day 12.From day 13,the mice were not given any treatment,and then the tumor volume of mice was measured every two days.When the tumor volume in control group increased to 2000 mm^(3)(day 24),the mice were killed to measure the volume of the transplanted tumor and calculate the tumor volume inhibition rate.Immunofluorescence staining was used to detect the expressions of neutrophil extracellular traps(NETs)-associated proteins citrullinated histone H3(CitH3) and myeloperoxidase(MPO) in transplanted tumour specimens.Blood was collected from the inner canthus of the eye,and the serum was centrifuged to detect the levels of tumor necrosis factor-α(TNF-α),and interferon-γ(IFN-γ) in mice by enzyme-linked immunosorbent assay(ELISA).Results Compared with control group,the transplanted tumour volume of mice in berberine group,irradiation group and berberine+irradiation group decreased(P<0.01),the fluorescence intensity of co-expressed CitH3 and MPO cells in berberine group decreased(P<0.01),and the expression levels of TNF-α and IFN-γ decreased(P<0.01),while the fluorescence intensity of co-expressed CitH3 and MPO cells and the expression levels of TNF-α and IF
作者
崔震庭
韩枫
段琼
汪鸣霄
马建新
CUI Zhenting;HAN Feng;DUAN Qiong;WANG Mingxiao;MA Jianxin(Department of Radiotherapy,Lianyungang Dongfang Hospital,Affiliated to Bengbu Medical College,Lianyungang 222042,China;Department of Radiotherapy,Lianyungang Dongfang Hospital Affiliated to Xuzhou Medical University;Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine)
出处
《山西医科大学学报》
CAS
2023年第12期1551-1555,共5页
Journal of Shanxi Medical University
基金
江苏省卫生计生委面上科研项目(H201671)。
