Non-alcoholic fatty liver disease(NAFLD)patients have multiple metabolic distur-bances,with markedly elevated levels of lactate.Lactate accumulations play pleiotropic roles in disease progression through metabolic rea...Non-alcoholic fatty liver disease(NAFLD)patients have multiple metabolic distur-bances,with markedly elevated levels of lactate.Lactate accumulations play pleiotropic roles in disease progression through metabolic rearrangements and epigenetic modifications.Mono-carboxylate transporter 4(MCT4)is highly expressed in hepatocytes and responsible for trans-porting intracellular lactate out of the cell.To explore whether elevated MCT4 levels played any role in NAFLD development,we overexpressed and silenced MCT4 in hepatocytes and per-formed a comprehensive in vitro and in vivo analysis.Our results revealed that MCT4 overex-pression down-regulated the genes for lipid synthesis while up-regulating the genes involved in lipid catabolism.Conversely,silencing MCT4 expression or inhibiting MCT4 expression led to the accumulation of intracellular lipid and glucose metabolites,resulting in hepatic steatosis.In a mouse model of NAFLD,we found that exogenous MCT4 overexpression significantly reduced lipid metabolism and alleviated hepatocellular steatosis.Mechanistically,MCT4 alle-viated hepatic steatosis by regulating a group of hub genes such as Arg2,Olr1,Cd74,Mmp8,Irf7,Spp1,and Apoe,which in turn impacted multiple pathways involved in lipid metabolism and inflammatory response,such as PPAR,HIF-1,TNF,IL-17,PI3K-AKT,Wnt,and JAK-STAT.Collectively,our results strongly suggest that MCT4 may play an important role in regulating lipid metabolism and inflammation and thus serve as a potential therapeutic target for NAFLD.展开更多
目的:研究追毒方对三阴性乳腺癌细胞MDA-MB-231增殖、侵袭迁移的影响,并阐述其分子机制。方法:首先采用MTT细胞毒性试验,Transwell小室侵袭试验和伤口愈合试验观察不同浓度追毒方对三阴性乳腺癌细胞增殖及侵袭迁移能力的影响;通过流式...目的:研究追毒方对三阴性乳腺癌细胞MDA-MB-231增殖、侵袭迁移的影响,并阐述其分子机制。方法:首先采用MTT细胞毒性试验,Transwell小室侵袭试验和伤口愈合试验观察不同浓度追毒方对三阴性乳腺癌细胞增殖及侵袭迁移能力的影响;通过流式细胞术和乳酸试剂盒评估不同浓度追毒方处理后三阴性乳腺癌细胞糖摄取及乳酸生成、外排水平,以观察其有氧糖酵解水平;Western blot法检测追毒方对三阴性乳腺癌细胞的细胞外基质金属蛋白酶诱导因子(CD147)糖基化及葡萄糖转运蛋白(GLUT1)、单羧酸转运蛋白(MCT4)表达的影响;为了进一步证明追毒方对TNBC细胞有氧糖酵解的调控与CD147糖基化有关,使用糖基转移酶GnT-V激动剂处理细胞,观察其对追毒方抑制效应的逆转作用。结果:与空白对照组相比,追毒方0.1、0.2、0.4、0.8、1.6、3.2μg/mL组的细胞增殖活性显著降低(P<0.01),24 h IC_(50)为5.00μg/mL,48 h IC_(50)为0.25μg/mL,72 h IC_(50)为0.02μg/mL,追毒方0.4、0.8、1.6μg/mL组和2-DG 2101μg/mL组的侵袭能力和迁移能力显著降低,CD147糖基化水平显著下降(P<0.01),糖摄取能力显著降低(P<0.01),细胞内和培养上清的乳酸含量显著降低(P<0.01),有氧糖酵解关键跨膜蛋白GLUT1/MCT4的表达显著下调(P<0.01)。视黄酸处理模型中,与空白对照组相比,视黄酸30μg/mL组的糖摄取能力、细胞内及培养上清乳酸含量,CD147糖基化水平降低,GLUT1/MCT4的表达明显上调(P<0.05或P<0.01);与追毒方0.8μg/mL组相比,视黄酸30μg/mL组+追毒方0.8μg/mL组的上述检测指标明显上调(P<0.05或P<0.01)。结论:追毒方通过调控CD147糖基化抑制GLUT1、MCT4表达,从而下调三阴性乳腺癌有氧糖酵解葡萄糖通量及乳酸生成和外排,最终抑制三阴性乳腺癌增殖及侵袭迁移。展开更多
基金supported in part by research grants from the Natural Science Foundation of China(No.82102696 to J.F.)the Chongqing Natural Science Foundation of China(No.2024NSCQ-MSX0073 to J.F.)+3 种基金the US National Institutes of Health(No.CA226303 to T.C.H.)supported in part by The University of Chicago Cancer Center Support Grant(No.P30CA014599)the National Center for Advancing Translational Sciences of the US National Institutes of Health(No.UL1 TR000430)T.C.H.was supported by the Mabel Green Myers Research Endowment Fund and The University of Chicago Orthopaedics Alumni Fund.
文摘Non-alcoholic fatty liver disease(NAFLD)patients have multiple metabolic distur-bances,with markedly elevated levels of lactate.Lactate accumulations play pleiotropic roles in disease progression through metabolic rearrangements and epigenetic modifications.Mono-carboxylate transporter 4(MCT4)is highly expressed in hepatocytes and responsible for trans-porting intracellular lactate out of the cell.To explore whether elevated MCT4 levels played any role in NAFLD development,we overexpressed and silenced MCT4 in hepatocytes and per-formed a comprehensive in vitro and in vivo analysis.Our results revealed that MCT4 overex-pression down-regulated the genes for lipid synthesis while up-regulating the genes involved in lipid catabolism.Conversely,silencing MCT4 expression or inhibiting MCT4 expression led to the accumulation of intracellular lipid and glucose metabolites,resulting in hepatic steatosis.In a mouse model of NAFLD,we found that exogenous MCT4 overexpression significantly reduced lipid metabolism and alleviated hepatocellular steatosis.Mechanistically,MCT4 alle-viated hepatic steatosis by regulating a group of hub genes such as Arg2,Olr1,Cd74,Mmp8,Irf7,Spp1,and Apoe,which in turn impacted multiple pathways involved in lipid metabolism and inflammatory response,such as PPAR,HIF-1,TNF,IL-17,PI3K-AKT,Wnt,and JAK-STAT.Collectively,our results strongly suggest that MCT4 may play an important role in regulating lipid metabolism and inflammation and thus serve as a potential therapeutic target for NAFLD.
文摘目的:研究追毒方对三阴性乳腺癌细胞MDA-MB-231增殖、侵袭迁移的影响,并阐述其分子机制。方法:首先采用MTT细胞毒性试验,Transwell小室侵袭试验和伤口愈合试验观察不同浓度追毒方对三阴性乳腺癌细胞增殖及侵袭迁移能力的影响;通过流式细胞术和乳酸试剂盒评估不同浓度追毒方处理后三阴性乳腺癌细胞糖摄取及乳酸生成、外排水平,以观察其有氧糖酵解水平;Western blot法检测追毒方对三阴性乳腺癌细胞的细胞外基质金属蛋白酶诱导因子(CD147)糖基化及葡萄糖转运蛋白(GLUT1)、单羧酸转运蛋白(MCT4)表达的影响;为了进一步证明追毒方对TNBC细胞有氧糖酵解的调控与CD147糖基化有关,使用糖基转移酶GnT-V激动剂处理细胞,观察其对追毒方抑制效应的逆转作用。结果:与空白对照组相比,追毒方0.1、0.2、0.4、0.8、1.6、3.2μg/mL组的细胞增殖活性显著降低(P<0.01),24 h IC_(50)为5.00μg/mL,48 h IC_(50)为0.25μg/mL,72 h IC_(50)为0.02μg/mL,追毒方0.4、0.8、1.6μg/mL组和2-DG 2101μg/mL组的侵袭能力和迁移能力显著降低,CD147糖基化水平显著下降(P<0.01),糖摄取能力显著降低(P<0.01),细胞内和培养上清的乳酸含量显著降低(P<0.01),有氧糖酵解关键跨膜蛋白GLUT1/MCT4的表达显著下调(P<0.01)。视黄酸处理模型中,与空白对照组相比,视黄酸30μg/mL组的糖摄取能力、细胞内及培养上清乳酸含量,CD147糖基化水平降低,GLUT1/MCT4的表达明显上调(P<0.05或P<0.01);与追毒方0.8μg/mL组相比,视黄酸30μg/mL组+追毒方0.8μg/mL组的上述检测指标明显上调(P<0.05或P<0.01)。结论:追毒方通过调控CD147糖基化抑制GLUT1、MCT4表达,从而下调三阴性乳腺癌有氧糖酵解葡萄糖通量及乳酸生成和外排,最终抑制三阴性乳腺癌增殖及侵袭迁移。
