目的 研究西黃丸組分中藥是否與西黃丸具有相似的抗癌作用及機(jī)制。方法 4T1乳腺癌細(xì)胞荷瘤建立動(dòng)物模型，西黃丸低、中、高劑量（0.39、0.78、1.95 g/kg）和西黃丸組分中藥0.3 mL （74.5 mg/kg欖香烯、2.73 mg/kg牛磺酸、0.52mg/kg麝香酮和4.75 mg/kg 11-羰基-β乙酯乳香酸混合物，對(duì)應(yīng)于西黃丸高劑量組） ig給藥14 d，剝離腫瘤組織，稱質(zhì)量，勻漿，制備單細(xì)胞懸液，免疫磁硃分離Treg細(xì)胞。流式細(xì)胞術(shù)和免疫組化檢測(cè)腫瘤微環(huán)境中Treg細(xì)胞數(shù)量變化情況，Western Blotting檢測(cè)腫瘤微環(huán)境中Treg細(xì)胞磷脂酰肌醇3-激酶/蛋白激酶B （PI3K/AKT）蛋白表達(dá)情況，實(shí)時(shí)熒光定量PCR （qRT-PCR）檢測(cè)腫瘤微環(huán)境中Treg細(xì)胞PI3K、AKT mRNA表達(dá)情況。結(jié)果 與模型組及西黃丸低、中劑量組比較，西黃丸組分中藥組腫瘤質(zhì)量顯著減少（P<0.05）；流式細(xì)胞術(shù)和免疫組化結(jié)果顯示，腫瘤微環(huán)境中Treg細(xì)胞數(shù)量顯著減少（P<0.05）；Western Blotting和qRT-PCR結(jié)果顯示，腫瘤微環(huán)境中Treg細(xì)胞PI3K、AKT蛋白及mRNA表達(dá)顯著下降（P<0.05）。與西黃丸高劑量組比較，西黃丸組分中藥組腫瘤質(zhì)量、Treg細(xì)胞數(shù)量和PI3K、AKT蛋白及mRNA表達(dá)均明顯升高（P<0.05）。結(jié)論 西黃丸組分中藥通過抑制腫瘤微環(huán)境中Treg細(xì)胞PI3K/AKT信號(hào)通路的表達(dá)減少Treg細(xì)胞數(shù)量，進(jìn)而表現(xiàn)與西黃丸相似的抗癌作用和機(jī)制。

Objective To study whether the Xihuang Pills component-based Chinese medicine has similar anti-cancer effect and mechanism as Xihuang Pills.Methods 4T1 breast cancer cells were used to establish a tumor model in mice.The model mice were ig administered with Xihuang Pills of low,medium,and high doses (0.39,0.78 and 1.95 g/kg) and Xihuang Pills component-based Chinese medicine of 0.3 mL (74.5 mg/kg elemene,2.73 mg/kg taurine,0.52 mg/kg Muscone and 4.75 mg/kg 11-carbonyl-beta-ethyl ester mastic acid mixture,corresponding to the high dosage group of Xihuang Pills) for 14 d.The tumor tissues were removed,weighed and homogenized to prepare single cell suspensions.Treg cells were prepared by magnetic cell sorting.The changes in number of Treg cells in the tumor microenvironment were detected by flow cytometry and immunohistochemistry.The protein expression of PI3K and AKT in Treg cells in the tumor microenvironment was detected by western blotting.The mRNA expression of PI3K and AKT in Treg cells in the tumor microenvironment was detected by the Real-time PCR (qRT-PCR).Results Compared with model group and Xihuang Pills low-and medium-dose groups,the weight of tumors was significantly decreased in the Xihuang Pills component-based Chinese medicine group (P< 0.05);The results of flow cytometry and immunohistochemistry showed a significant decrease in the number of Treg cells in the tumor microenvironment (P< 0.05);Western Blotting and qRT-PCR results showed that the expression of PI3K,AKT protein and their mRNA in Treg cells in the tumor microenvironment was significantly decreased (P< 0.05).Compared with the Xihuang Pills high-dose group,the tumor weight,the number of Treg cells,and the protein expression of PI3K and AKT and their mRNA expression in the Xihuang Pills component-based Chinese medicine group were significant increased (P< 0.05).Conclusion Xihuang Pills component-based Chinese medicine can reduce the number of Treg cells by inhibiting the expression of PI3K/AKT signaling pathway in Treg cells in the tumor microenvironment,and thus exhibit similar anti-cancer effects and mechanisms as Xihuang Pills.

國家自然科學(xué)基金資助項(xiàng)目（81573664）