论文部分内容阅读
目的探讨中药升清胶囊通过干预CXCL12-CXCR4信号通路对脂代谢异常的调节及其对乳腺癌肝转移的影响。方法(1)选用人乳腺癌细胞株MDA-MB-231,分为对照组(Con)、空白血清组(Black)、表阿霉素组(Epirubicin)以及升清胶囊血清低浓度组(SQ-L)、高浓度组(SQ-H)。采用MTT法检测药物血清对细胞增殖的影响,并用划痕愈合实验和Transwell实验检测其对细胞迁移及侵袭能力的影响。同时采用Western blotting法检测CXC趋化因子配体12(CXCL12)与其特异性受体CXCR4以及蛋白激酶B(AKT),磷酸化蛋白激酶B(p-AKT),B淋巴白血病-淋巴瘤-2基因(Bcl-2),基质金属蛋白酶9(MMP-9)蛋白表达水平的变化。(2)建立脂代谢异常乳腺癌肝转移动物裸鼠模型,用升清胶囊0.25 g·kg~(-1),或表阿霉素5μg·g~(-1)干预治疗,检测血浆游离脂肪酸(FFA)、三酰甘油(TG)水平变化及对乳腺癌肝转移的影响。结果 (1)MTT结果表明,升清胶囊和表阿霉素药物血清干预后,MDA-MB-231细胞存活率显著降低,且呈浓度依赖性持续降低(P<0.05)。Transwell和划痕愈合实验结果表明,升清胶囊药物血清能够显著抑制细胞的侵袭和迁移能力。Western blotting结果表明,与对照组比较,升清胶囊药物可降低趋化因子CXCL12以及受体CXCR4蛋白表达水平(P<0.05),同时下调MMP-9和Bcl-2蛋白表达,且p-AKT水平降低(P<0.05)。(2)动物实验表明,升清胶囊可以显著降低脂代谢异常裸鼠TG、FFA水平(P<0.05),抑制乳腺癌肝转移。结论升清胶囊能够通过抑制趋化因子CXCL12-CXCR4和相关信号通路活性,下调血脂水平,抑制乳腺癌转移。
Objective To investigate the regulation of lipid metabolic abnormalities by Chinese medicine Shengqing capsule on CXCL12-CXCR4 signaling pathway and its effect on liver metastasis of breast cancer. Methods (1) Human breast cancer cell line MDA-MB-231 was selected and divided into control group, blank group, epirubicin group and SQ- L), high concentration group (SQ-H). MTT assay was used to detect the effect of drug serum on cell proliferation. Scratch healing assay and Transwell assay were used to detect the effect of drug serum on cell migration and invasion. Meanwhile, the expressions of CXCL12 and CXCR4, AKT, p-AKT, B lymphoblastic leukemia-2 gene were detected by Western blotting. (Bcl-2), matrix metalloproteinase 9 (MMP-9) protein expression levels. (2) To establish the animal model of hepatic metastasis of breast cancer with lipid metabolism abnormalities, and to treat the liver metastasis of nude mice with lipid metabolism abnormalities. The serum free fatty acid was detected with Shengqing capsule 0.25 g · kg -1 or epirubicin 5 μg · g -1. (FFA), triglyceride (TG) levels and liver metastasis of breast cancer. Results (1) The results of MTT showed that the survival rate of MDA-MB-231 cells was significantly decreased after the intervention of Shengqing Capsule and epirubicin serum (P <0.05). Transwell and wound healing experiments show that Shengqing Capsule serum can significantly inhibit cell invasion and migration. Western blotting results showed that compared with the control group, Shengqing Capsule could decrease the expression of chemokine CXCL12 and CXCR4 (P <0.05), and down-regulate the expressions of MMP-9 and Bcl-2, Decreased (P <0.05). (2) Animal experiments show that Shengqing Capsule can significantly reduce lipid metabolism in nude mice TG, FFA levels (P <0.05), inhibit liver metastases of breast cancer. Conclusion Shengqing Capsule can inhibit the chemokine CXCL12-CXCR4 and related signal pathway activity, down-regulation of blood lipid levels, inhibition of breast cancer metastasis.