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纸质出版日期:2018
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杨丽霞, 邓卫芳, 陈鸿, 等. 蓝萼香茶菜二萜类物质对乳腺癌癌前病变肝郁肾虚证模型大鼠的干预作用[J]. 中国实验方剂学杂志, 2018,24(2):140-145.
YANG Li-xia, DENG Wei-fang, CHEN Hong, et al. Intervention Effect of var. Diterpenoids on Liver Qi Stagnation and Kidney Deficiency Model of Precancerous Breast Lesions[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(2): 140-145.
杨丽霞, 邓卫芳, 陈鸿, 等. 蓝萼香茶菜二萜类物质对乳腺癌癌前病变肝郁肾虚证模型大鼠的干预作用[J]. 中国实验方剂学杂志, 2018,24(2):140-145. DOI: 10.13422/j.cnki.syfjx.2018020140.
YANG Li-xia, DENG Wei-fang, CHEN Hong, et al. Intervention Effect of var. Diterpenoids on Liver Qi Stagnation and Kidney Deficiency Model of Precancerous Breast Lesions[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(2): 140-145. DOI: 10.13422/j.cnki.syfjx.2018020140.
目的:通过加热回流的方法提取蓝萼香茶菜中二萜类物质,测定其含量,研究二萜类物质对乳腺癌癌前病变肝郁肾虚证模型大鼠的干预作用及其作用机制。方法:雌性SD大鼠随机分层分组为正常组、枸橼酸他莫昔芬片组、模型组、蓝萼组。除正常组外其他各组均用二甲基苯蒽(DMBA)结合慢性不可预见应激行为制备乳腺癌癌前病变肝郁肾虚证模型,造模结束后,观察记录各组大鼠的生物表征变化及病理变化验证模型。蓝萼组给予二萜类提取物水溶液5 mg·kg-1灌胃,正常组和模型组给予蒸馏水10 mL·kg-1灌胃,各组均连续灌胃30 d。30 d后,记录各组大鼠肉眼可见肿瘤的发生情况;腹腔取血,使用酶联免疫吸附试验(ELISA)试剂盒检测血清中雌二醇(E2),孕激素(P)水平;采集乳腺织织,用苏木素-伊红(HE)染色观察乳腺组织病理学变化,实时荧光定量PCR(Real-time PCR)及蛋白免疫印迹法(Western blot)检测乳腺组织血管内皮生长因子(VEGF),趋化因子受体4(CXCR4)mRNA及蛋白表达水平。结果:提取得干燥品质量2.162 g,干燥品中二萜类质量分数为26.35%。与模型组比较,蓝萼组血清中E2,P水平明显降低(P<0.05),乳腺组织中VEGF,CXCR4蛋白及mRNA表达水平均明显降低(P<0.05)。结论:蓝萼香茶菜二萜类物质有可能通过抑制VEGF,CXCR4蛋白及mRNA表达,抑制血管生成和趋化因子受体表达,调节大鼠激素水平,从而抑制大鼠乳腺癌癌前病变癌变的发生。
Objective:To extract Rabdosia japonica var. glaucocalyx diterpenes by heating reflux to determine the contents of diterpenes and study the effect of R. japonica var. glaucocalyx diterpenes on the liver Qi stagnation and kidney deficiency model of precancerous breast lesions. Method:Female SD rats were randomly divided into 4 groups
namely normal group
tamoxifen citrate group
model group
and R. japonica var. glaucocalyx diterpenes group. All of the groups except for normal group were given DMBA combined with multi-composite stress. Changes in biological characterizations of rats were observed and recorded after successful modeling. Diterpenes were given to R. japonica var. glaucocalyx diterpenes group
while distilled water was given to normal group and model group. Intragastric administration was performed for consecutively 30 days. Thirty days later
blood was collected from abdominal cavity and breast tissues for pathological examination. Incidence of tumors was recorded. Serum estradiol (E2) and progesterone (P) levels were detected by enzyme-linked immunosorbent assay (ELISA). Hematoxylin-eosin (HE) staining was adopted for observing the pathological changes. Expressions of vascular endothelial growth factor (VEGF)
chemokine receptor (CXCR4) protein and mRNA were detected by Real-time PCR and Western blot. Result:The dried products weighed 2.162 g
including 26.35% of diterpenes. Compared with normal group
a series of biological changes were observed in model group. The levels of E2 and P in R. japonica var. glaucocalyx diterpenes group were lower than those of model group (P<0.05). VEGF and CXCR4 protein and mRNA expressions were lower than those in model group(P<0.05). Conclusion:R. japonica var. glaucocalyx diterpenes can prohibit VEGF and CXCR4 mRNA and protein expressions
angiogenesis and chemokine receptor expressions
and regulate hormone levels in rats
so as to inhibit the carcinogenesis of precancerous breast lesions in rats.
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