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该研究拟从体内角度研究半枝莲总黄酮(TF-SB)的抗肿瘤作用,并探讨TF-SB对肿瘤细胞自噬的影响及其对PI3K/AKT/mTOR通路的调控作用,为TF-SB的抗肿瘤作用提供实验依据。实验以黑色素瘤荷瘤小鼠模型为研究对象,分为空白对照组、雷帕霉素组(Rap,1.5 mg·kg~(-1))、TF-SB高、中、低剂量组(200,100,50 mg·kg~(-1)),每天给药1次,连续用药11 d,通过测量肿瘤的体积及抑瘤率,来观察TF-SB对黑色素瘤生长的抑制效果;通过TUNEL法检测肿瘤细胞凋亡情况,来进一步验证TF-SB的抗肿瘤活性;通过Western blot法检测LC3-Ⅰ,LC3-Ⅱ的蛋白表达,计算LC3-Ⅱ对LC3-Ⅰ的相对表达量,探讨TF-SB对肿瘤细胞自噬的诱导作用;通过检测PI3K/AKT/mTOR通路标志蛋白的磷酸化水平,研究TF-SB诱导细胞自噬的分子机制。结果表明,TF-SB可有效抑制荷瘤小鼠体内黑色素瘤的生长,减小肿瘤体积、提高抑瘤率,并且可以显著增加肿瘤细胞凋亡指数,增加LC3-Ⅱ/LC3-Ⅰ,抑制p-PI3K,p-AKT及p-mTOR的蛋白表达水平,与对照组比较有显著性差异(P<0.05,P<0.01或P<0.001)。由此可见,半枝莲总黄酮具有抑制体内黑色素瘤生长的作用,其机制可能与通过抑制PI3K/AKT/mTOR通路,诱导肿瘤细胞自噬及凋亡有关。
The aim of this study was to investigate the anti-tumor effect of Scutellaria barbata flavonoids (TF-SB) in vivo and to explore the effect of TF-SB on the autophagy of tumor cells and the regulation of PI3K / AKT / mTOR pathway. The antitumor effect of SB provides experimental evidence. The experiment was conducted on the melanoma tumor-bearing mouse model and divided into blank control group, Rapamycin group (Rap, 1.5 mg · kg -1), TF-SB high, middle and low dose group , 50 mg · kg ~ (-1)) once a day for 11 days. The inhibitory effect of TF-SB on the growth of melanoma was observed by measuring the tumor volume and tumor inhibition rate. By TUNEL assay To further verify the antitumor activity of TF-SB, the protein expression of LC3-Ⅰ and LC3-Ⅱ was detected by Western blot, and the relative expression level of LC3-Ⅱ and LC3- On the induction of autophagy in tumor cells. The molecular mechanism of TF-SB-induced autophagy was investigated by detecting the phosphorylation of PI3K / AKT / mTOR pathway marker proteins. The results showed that TF-SB could effectively inhibit the growth of melanoma in tumor-bearing mice, reduce the tumor volume, increase the tumor inhibition rate, and significantly increase the apoptosis index of tumor cells, increase LC3-Ⅱ / LC3-Ⅰ and inhibit p -PI3K, p-AKT and p-mTOR protein expression levels compared with the control group were significantly different (P <0.05, P <0.01 or P <0.001). Thus, Scutellaria barbata total flavonoids can inhibit the growth of melanoma in vivo, the mechanism may be related to inhibition of PI3K / AKT / mTOR pathway to induce tumor cell autophagy and apoptosis.