發(fā)布時(shí)間：2019-06-06 10:06

【摘要】：目的建立氣虛血瘀證乳腺癌荷瘤4-T1小鼠模型,并予益氣活血中藥干預(yù),分析在氣虛血瘀狀態(tài)下中藥干預(yù)對(duì)腫瘤微環(huán)境的影響。方法雌性BALB/c小鼠48只,隨機(jī)平均分為6組:空白對(duì)照組、單純荷瘤組、氣虛血瘀證荷瘤模型組(復(fù)合模型組)、復(fù)合模型低劑量組、復(fù)合模型中劑量組、復(fù)合模型高劑量組。單純荷瘤組小鼠注射4T1單細(xì)胞懸液,復(fù)合模型組在單純荷瘤組基礎(chǔ)上注射利血平。從實(shí)驗(yàn)第15天起,復(fù)合模型低劑量組、中劑量組、高劑量組分別灌胃相應(yīng)劑量中藥(四君子湯合血府逐瘀湯),每日1次,連續(xù)28 d。分別于實(shí)驗(yàn)第7、14、21、28、35、42天對(duì)復(fù)合模型組及各中藥干預(yù)組予癥狀量化評(píng)分表評(píng)估,第42天測(cè)量各組耳緣、腳趾、腹壁靜脈微循環(huán)血流量,Western blot法測(cè)定腫瘤組織基質(zhì)金屬蛋白酶2(MMP2)、磷酸化細(xì)胞外信號(hào)調(diào)節(jié)激酶(p-ERK1/2)的表達(dá)。結(jié)果復(fù)合模型組、復(fù)合模型低劑量組、復(fù)合模型中劑量組、復(fù)合模型高劑量組均出現(xiàn)氣虛血瘀癥狀,低劑量組量化評(píng)分值自第28天低于復(fù)合模型組(P0.05),中劑量組自第21天分值低于復(fù)合模型組(P0.05)�？瞻讓�(duì)照組靜脈微循環(huán)血流量大于其余各組(P0.05),復(fù)合模型組小于單純荷瘤組(P0.05),各中藥干預(yù)組均比復(fù)合模型組提高(P0.01或P0.05)。復(fù)合模型組原位瘤MMP-2表達(dá)高于單純荷瘤組(P0.05),各中藥干預(yù)組均低于復(fù)合模型組(P0.05)。復(fù)合模型低劑量組、中劑量組原位瘤p-ERK1/2表達(dá)均低于復(fù)合模型組(P0.05或P0.01)。結(jié)論通過(guò)注射4T1單細(xì)胞懸液結(jié)合注射利血平方法建立的氣虛血瘀證乳腺癌小鼠模型穩(wěn)定可靠,符合中醫(yī)證候特點(diǎn)。氣虛血瘀證小鼠腫瘤組織內(nèi)MMP-2、p-ERK1/2表達(dá)上調(diào),益氣活血中藥干預(yù)后能糾正氣虛血瘀狀態(tài)同時(shí)下調(diào)MMP-2、p-ERK1/2的表達(dá),抑制腫瘤生長(zhǎng)。
[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.
【作者單位】： 浙江中醫(yī)藥大學(xué)附屬第二醫(yī)院全科醫(yī)學(xué)科;浙江中醫(yī)藥大學(xué)附屬第一醫(yī)院腫瘤科;浙江中醫(yī)藥大學(xué);
【基金】：浙江省自然科學(xué)基金項(xiàng)目(No.Y13H290038) 浙江省中醫(yī)藥科學(xué)研究基金計(jì)劃項(xiàng)目(No.2012ZA049)~~
【分類(lèi)號(hào)】：R285.5;R-332

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