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半夏泻心汤重塑色氨酸代谢抑制结肠癌进展的效应及机制研究 被引量:1
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作者 蒋义芳 黄渝清 +4 位作者 赖恒周 李雪珂 龙柳伊 由凤鸣 旷歧轩 《中国中药杂志》 北大核心 2025年第5期1310-1320,共11页
探究半夏泻心汤通过重塑色氨酸代谢抑制结肠癌进展的效应及机制。将Balb/c小鼠分为对照(control)组、模型(model)组、半夏泻心汤低剂量(BXD-L)组和半夏泻心汤高剂量(BXD-H)组。除control组外,其余各组皮下注射CT26-Luc细胞建立结肠癌模... 探究半夏泻心汤通过重塑色氨酸代谢抑制结肠癌进展的效应及机制。将Balb/c小鼠分为对照(control)组、模型(model)组、半夏泻心汤低剂量(BXD-L)组和半夏泻心汤高剂量(BXD-H)组。除control组外,其余各组皮下注射CT26-Luc细胞建立结肠癌模型,并予以半夏泻心汤进行干预。采用小动物活体成像监测肿瘤生长情况,并测量肿瘤体积和质量;苏木精-伊红(HE)染色法观察小鼠肿瘤组织病理改变;采用免疫组化法检测肿瘤组织Ki67表达;免疫荧光法和流式细胞术检测肿瘤组织中CD3^(+)/CD8^(+)T细胞浸润情况和数目变化,酶联免疫吸附测定(ELISA)法检测肿瘤组织中干扰素-γ(IFN-γ)和白细胞介素2(IL-2)含量变化;靶向代谢组学检测血清中色氨酸(Trp)代谢变化,并测量肿瘤组织中色氨酸含量,采用蛋白免疫印迹(Western blot)法和实时荧光定量PCR(RT-qPCR)法检测肿瘤组织中吲哚胺2,3-双加氧酶1(IDO1)、原癌基因MYC、溶质载体基因家族7成员5(SLC7A5)蛋白及mRNA水平变化。同时,在体外建立“CT26结肠癌细胞+CD8^(+)T细胞”共培养模型,予以半夏泻心汤含药血清干预,采用细胞增殖与活性检测-8(CCK-8)法检测CT26细胞活性,分别测量CT26细胞和CD8^(+)T细胞Trp含量、CD8^(+)T细胞IFN-γ和IL-2的分泌情况;采用RT-qPCR法检测CT26细胞中MYC、SLC7A5 mRNA水平变化。结果显示半夏泻心汤干预后,可明显抑制小鼠结肠癌肿瘤增长,减轻瘤质量、缩小肿瘤体积;组织病理学显示肿瘤细胞排列稀疏,且有不同程度的片状坏死区,且肿瘤组织中增殖标志物Ki67水平表达降低;半夏泻心汤可升高肿瘤组织中IFN-γ、IL-2含量,同时上调CD3^(+)/CD8^(+)T细胞比例,并且降低Trp、IDO1、MYC和SLC7A5含量。体外共培养实验表明半夏泻心汤含药血清可降低CT26细胞对Trp的摄取、增加CD8^(+)T细胞Trp含量,并增强CD8^(+)T细胞IL-2、IFN-γ的分泌,同时降低CT26细胞中MYC、SLC7A5 mRNA含量。综上所述,半夏泻心汤能够抑制MYC/SLC7A5通路重塑结肠癌Trp代谢,调复CD8^(+)T细胞对Trp摄取而增加其细胞毒性,从而抑制结肠癌发展。 展开更多
关键词 半夏泻心汤 结肠癌 CD8^(+)T细胞 色氨酸代谢 myc/slc7a5通路
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BmSLC7A5 is essential for silk protein synthesis and larval development in Bombyx mori
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作者 Xin Tang Huawei Liu +4 位作者 Xin Wang Li Chang Qingsong Liu Qingyou Xia Ping Zhao 《Insect Science》 SCIE CAS CSCD 2024年第5期1425-1439,共15页
Insects produce silk to form cocoons,nests,and webs,which are important for their survival and reproduction.However,little is known about the molecular mecha-nism of silk protein synthesis at the translation level.The... Insects produce silk to form cocoons,nests,and webs,which are important for their survival and reproduction.However,little is known about the molecular mecha-nism of silk protein synthesis at the translation level.The solute carrier family 7(SLC7)genes are involved in activating the target of rapamycin complex 1(TORC1)signaling pathway and protein translation process,but the physiological roles of SLC7 genes in silk-producing insects have not been reported.Here,we found that amino acid signaling regulates silk protein synthesis and larval development via the L-type amino acid trans-porter 1(LAT1;also known as SLC7A5)in Bombyx mori.A total of 12 SLC7 homologs were identified in the silkworm genome,among which BmSLC7A5 was found to be a silk gland-enriched gene and may be involved in leucine transport.Bioinformatics analy-sis indicated that SLC7A5 displays high homology and a close phylogenetic relationship in silk-producing insects.Subsequently,we found that leucine treatment significantly in-creased silk protein synthesis by improving the transcription and protein levels of silk genes.Furthermore,systemic and silk gland-specific knockout of BmSLC7A5 led to de-creased silk protein synthesis by inhibiting TORC1 signaling,and somatic mutation also resulted in arrested development from the 5th instar to the early pupal stage.Altogether,our study reveals that BmSLC7A5 is involved in regulating silk protein synthesis and larval development by affecting the TORC1 signaling pathway,which provides a new strategy and target for improving silk yield. 展开更多
关键词 amino acid signaling Bombyx mori larval development slc7a5 silk pro-tein synthesis TORC1 signaling pathway
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