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乳腺癌气虚血瘀证小鼠模型的建立及益气活血法的干预研究

发布时间:2019-06-06 10:06
【摘要】:目的建立气虚血瘀证乳腺癌荷瘤4-T1小鼠模型,并予益气活血中药干预,分析在气虚血瘀状态下中药干预对肿瘤微环境的影响。方法雌性BALB/c小鼠48只,随机平均分为6组:空白对照组、单纯荷瘤组、气虚血瘀证荷瘤模型组(复合模型组)、复合模型低剂量组、复合模型中剂量组、复合模型高剂量组。单纯荷瘤组小鼠注射4T1单细胞悬液,复合模型组在单纯荷瘤组基础上注射利血平。从实验第15天起,复合模型低剂量组、中剂量组、高剂量组分别灌胃相应剂量中药(四君子汤合血府逐瘀汤),每日1次,连续28 d。分别于实验第7、14、21、28、35、42天对复合模型组及各中药干预组予症状量化评分表评估,第42天测量各组耳缘、脚趾、腹壁静脉微循环血流量,Western blot法测定肿瘤组织基质金属蛋白酶2(MMP2)、磷酸化细胞外信号调节激酶(p-ERK1/2)的表达。结果复合模型组、复合模型低剂量组、复合模型中剂量组、复合模型高剂量组均出现气虚血瘀症状,低剂量组量化评分值自第28天低于复合模型组(P0.05),中剂量组自第21天分值低于复合模型组(P0.05)。空白对照组静脉微循环血流量大于其余各组(P0.05),复合模型组小于单纯荷瘤组(P0.05),各中药干预组均比复合模型组提高(P0.01或P0.05)。复合模型组原位瘤MMP-2表达高于单纯荷瘤组(P0.05),各中药干预组均低于复合模型组(P0.05)。复合模型低剂量组、中剂量组原位瘤p-ERK1/2表达均低于复合模型组(P0.05或P0.01)。结论通过注射4T1单细胞悬液结合注射利血平方法建立的气虚血瘀证乳腺癌小鼠模型稳定可靠,符合中医证候特点。气虚血瘀证小鼠肿瘤组织内MMP-2、p-ERK1/2表达上调,益气活血中药干预后能纠正气虚血瘀状态同时下调MMP-2、p-ERK1/2的表达,抑制肿瘤生长。
[Abstract]:Objective To establish a 4-T1 mouse model of breast cancer with Qi-deficiency and blood stasis syndrome, and to treat the tumor microenvironment by using the traditional Chinese medicine in the condition of qi-deficiency and blood stasis. Methods Forty-eight female BALB/ c mice were randomly divided into 6 groups: the blank control group, the pure tumor-bearing group, the Qi-deficiency and blood-stasis syndrome-bearing model group (compound model group), the low-dose group of the composite model, the dose group in the composite model, and the high-dose group of the compound model. The mice were injected with 4T1 single cell suspension, and the compound model group was injected with reserpine on the basis of the simple tumor-bearing group. From the 15th day of the experiment, the low-dose group, middle-dose group and high-dose group of the compound model were given the corresponding dose of the traditional Chinese medicine (Sijunzi Decoction and Xuefu Zhuyu Decoction), once a day, for 28d, respectively. The expression of matrix metalloproteinase-2 (MMP2) and extracellular signal-regulated kinase (p-ERK1/2) was determined by Western blot. Results The combined model group, the low-dose group of the compound model, the dose group in the compound model, the high-dose group of the compound model had the symptoms of Qi deficiency and blood stasis, and the quantitative score of the low-dose group was lower than that of the compound model group at the 28th day (P0.05), and the score of the middle-dose group was lower than that of the compound model group (P0.05). The blood flow of the vein in the blank control group was greater than that of the other groups (P0.05). The combined model group was less than that of the simple tumor-bearing group (P0.05). The intervention group of the traditional Chinese medicine group was higher than that of the compound model group (P 0.01 or P0.05). The expression of MMP-2 in the compound model group was higher than that of the simple tumor-bearing group (P0.05). In the low-dose group, the expression of p-ERK1/2 in the middle-dose group was lower than that of the compound model group (P0.05 or P0.01). Conclusion The model of breast cancer with Qi-deficiency and blood stasis syndrome established by injection of 4T1 single cell suspension in combination with the injection of reserpine was stable and reliable, and it was in accordance with the characteristics of TCM syndrome. The expression of MMP-2 and p-ERK1/2 in the tumor tissue of mice with Qi-deficiency and blood stasis syndrome was up-regulated, and the expression of MMP-2 and p-ERK1/2 was down-regulated in the condition of qi-deficiency and blood stasis, and the growth of the tumor was inhibited.
【作者单位】: 浙江中医药大学附属第二医院全科医学科;浙江中医药大学附属第一医院肿瘤科;浙江中医药大学;
【基金】:浙江省自然科学基金项目(No.Y13H290038) 浙江省中医药科学研究基金计划项目(No.2012ZA049)~~
【分类号】:R285.5;R-332

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