目的探讨大脑中动脉(middle cerebral artery,MCA)狭窄性脑梗死患者血清血管生成素样蛋白4(angiopoietin-like protein 4,ANGPTL4)、CXC族趋化因子配体12(CXC chemokine ligand 12,CXCL12)、酰基辅酶A合成酶长链家族成员4(acyl CoA synt...目的探讨大脑中动脉(middle cerebral artery,MCA)狭窄性脑梗死患者血清血管生成素样蛋白4(angiopoietin-like protein 4,ANGPTL4)、CXC族趋化因子配体12(CXC chemokine ligand 12,CXCL12)、酰基辅酶A合成酶长链家族成员4(acyl CoA synthetase long-chain family member 4,ACSL4)水平的变化趋势及对患者预后的预测效能。方法选取2022年4月至2025年4月开封市第二中医院收治的152例MCA狭窄性脑梗死患者进行前瞻性研究,依据3个月时的预后,将患者分别纳入良好组(86例)、不良组(66例)。比较两组不同治疗时间血清ANGPTL4、CXCL12、ACSL4水平、收缩期峰值流速(peak systolic velocity,PSV)值、梗死体积及变化值,分析血清ANGPTL4、CXCL12、ACSL4水平变化值与PSV、梗死体积变化值的相关性,分析治疗7、14 d后血清ANGPTL4、CXCL12、ACSL4水平变化值对患者预后的预测效能。结果随着治疗时间的延长,两组ANGPTL4水平降低,血清CXCL12与ACSL4水平、PSV值、梗死体积均升高,差异有统计学意义(P<0.05);治疗7、14 d后,不良组血清ANGPTL4水平低于良好组,血清CXCL12、ACSL4水平、PSV值、梗死体积高于良好组,差异有统计学意义(P<0.05);不良组治疗7、14 d后血清ANGPTL4、CXCL12、ACSL4水平、PSV值、梗死体积变化值低于良好组,差异有统计学意义(P<0.05);血清ANGPTL4、CXCL12、ACSL4水平变化值与PSV值、梗死体积及变化值均呈正相关,差异有统计学意义(P<0.05);治疗7、14 d后血清ANGPTL4、CXCL12、ACSL4水平变化值联合预测患者预后的AUC值高于各指标单独检测,差异有统计学意义(P<0.05)。结论血清ANGPTL4、CXCL12、ACSL4水平在MCA狭窄性脑梗死患者治疗过程中均呈现明显变化趋势,治疗7、14 d后,各指标变化值联合可为临床预测MCA狭窄性脑梗死患者预后提供参考。展开更多
Ketosis,a common metabolic disease during early lactation,is associated with high circulating levels ofβ-hydroxybutyrate(BHB).A portion of BHB that reaches the mammary gland is utilized as precursor for synthesis of ...Ketosis,a common metabolic disease during early lactation,is associated with high circulating levels ofβ-hydroxybutyrate(BHB).A portion of BHB that reaches the mammary gland is utilized as precursor for synthesis of fatty acids.Recent findings from nonruminant studies revealed that long chain fatty acyl-CoA ligase 4(ACSL4)could play a role in the regulation of cellular fatty acid metabolism,but the mechanisms by which ACSL4 mediates cellular lipid metabolism in response to BHB remains unclear.To achieve the aims,we conducted in vivo or in vitro analyses using bovine mammary gland biopsies and the immortalized mammary epithelial cell line(MAC-T).The in vivo study(n=6 cows per group)involved healthy cows(plasma BHB<0.60 mmol L^(–1))or ketotic cows(plasma BHB>2.0 mmol L^(–1))from which mammary gland tissue was biopsied.In vitro,MAC-T cells were challenged with 0,0.3,0.6,1.2,or 2.4 mmol L^(–1)BHB for 24 h to determine an optimal dose.Subsequently,MAC-T were incubated with 1.2 mmol L^(–1)BHB for 0,3,6,12,24,or 48 h.Furthermore,MAC-T cells were treated with small interfering ACSL4(siACSL4)for 24 h or ACSL4 overexpression plasmid(pcACSL4)for 36 h followed by a challenge with 1.2 mmol L^(–1)BHB for 24 h.Results showed that increased mRNA and protein abundance of lipogenic genes were linked to both mammary gland and in vitro challenge with BHB.BHB increased fatty acid content by activating ACSL4 expression,whereas inhibition of ACSL4 reduced BHB-induced reactive oxygen species(ROS)overproduction,enhancement of mitochondrial membrane potential,increase in fatty acid content,and lipid droplet accumulation.Furthermore,we also elevated ACSL4 expression with an overexpression plasmid to clarify its molecular role in response to BHB challenge.ACSL4 overexpression enhances BHB-induced lipid droplet accumulation by increased fatty acid content.Overall,the information showed that ACSL4 is crucial for the process of producing fatty acids from exogenous BHB.Reduced ACSL4 decreased fatty acid content and lipid droplet accumulation,improved mitochondrial function,directed more fatty acids towards oxidation.Thus,ACSL4 plays an important role in determining the fate of intracellular fatty acids and BHB in BMECs.展开更多
目的研究拟探讨瘦素(leptin)对长链脂酰辅酶A家族成员长链脂酰辅A合成酶5(acyl-CoA synthetase long chain family member 5,ACSL5)的调控作用及机制,并探究其对乳腺癌细胞迁移和侵袭的影响。方法免疫荧光实验检测瘦素受体的表达;划痕...目的研究拟探讨瘦素(leptin)对长链脂酰辅酶A家族成员长链脂酰辅A合成酶5(acyl-CoA synthetase long chain family member 5,ACSL5)的调控作用及机制,并探究其对乳腺癌细胞迁移和侵袭的影响。方法免疫荧光实验检测瘦素受体的表达;划痕愈合实验和Transwell实验检测细胞迁移、侵袭能力;采用PCR芯片数据分析瘦素下游靶基因,生物信息学方法分析ACSL5在乳腺癌中的表达、与患者分期及预后关系;RT-qPCR检测不同浓度瘦素处理后ACSL5的表达;慢病毒转染构建过表达ACSL5稳转株;Western blot检测上皮-间质转化(epithelial-mesenchymal transition,EMT)相关蛋白表达及AMPK-α、p-AMPK-α的表达。结果瘦素促进MCF-7细胞体外迁移和侵袭及EMT;ACSL5是瘦素下游靶基因,在乳腺癌中显著低表达且与患者预后相关;瘦素通过其受体OBR下调ACSL5的表达;瘦素激活AMPK通路下调ACSL5促进乳腺癌MCF-7细胞迁移和侵袭及EMT。结论瘦素可能通过激活AMPK通路下调ACSL5,增强乳腺癌细胞的迁移和侵袭能力并促进其EMT进程,从而促进乳腺癌恶性进展。展开更多
基金supported by the grants from the National Key Research and Development Program of China(2023YFD1800804 and 2023YFD1801100)the National Natural Science Foundation of China(32172926)the China Agriculture Research System(CARS-36)。
文摘Ketosis,a common metabolic disease during early lactation,is associated with high circulating levels ofβ-hydroxybutyrate(BHB).A portion of BHB that reaches the mammary gland is utilized as precursor for synthesis of fatty acids.Recent findings from nonruminant studies revealed that long chain fatty acyl-CoA ligase 4(ACSL4)could play a role in the regulation of cellular fatty acid metabolism,but the mechanisms by which ACSL4 mediates cellular lipid metabolism in response to BHB remains unclear.To achieve the aims,we conducted in vivo or in vitro analyses using bovine mammary gland biopsies and the immortalized mammary epithelial cell line(MAC-T).The in vivo study(n=6 cows per group)involved healthy cows(plasma BHB<0.60 mmol L^(–1))or ketotic cows(plasma BHB>2.0 mmol L^(–1))from which mammary gland tissue was biopsied.In vitro,MAC-T cells were challenged with 0,0.3,0.6,1.2,or 2.4 mmol L^(–1)BHB for 24 h to determine an optimal dose.Subsequently,MAC-T were incubated with 1.2 mmol L^(–1)BHB for 0,3,6,12,24,or 48 h.Furthermore,MAC-T cells were treated with small interfering ACSL4(siACSL4)for 24 h or ACSL4 overexpression plasmid(pcACSL4)for 36 h followed by a challenge with 1.2 mmol L^(–1)BHB for 24 h.Results showed that increased mRNA and protein abundance of lipogenic genes were linked to both mammary gland and in vitro challenge with BHB.BHB increased fatty acid content by activating ACSL4 expression,whereas inhibition of ACSL4 reduced BHB-induced reactive oxygen species(ROS)overproduction,enhancement of mitochondrial membrane potential,increase in fatty acid content,and lipid droplet accumulation.Furthermore,we also elevated ACSL4 expression with an overexpression plasmid to clarify its molecular role in response to BHB challenge.ACSL4 overexpression enhances BHB-induced lipid droplet accumulation by increased fatty acid content.Overall,the information showed that ACSL4 is crucial for the process of producing fatty acids from exogenous BHB.Reduced ACSL4 decreased fatty acid content and lipid droplet accumulation,improved mitochondrial function,directed more fatty acids towards oxidation.Thus,ACSL4 plays an important role in determining the fate of intracellular fatty acids and BHB in BMECs.
文摘目的研究拟探讨瘦素(leptin)对长链脂酰辅酶A家族成员长链脂酰辅A合成酶5(acyl-CoA synthetase long chain family member 5,ACSL5)的调控作用及机制,并探究其对乳腺癌细胞迁移和侵袭的影响。方法免疫荧光实验检测瘦素受体的表达;划痕愈合实验和Transwell实验检测细胞迁移、侵袭能力;采用PCR芯片数据分析瘦素下游靶基因,生物信息学方法分析ACSL5在乳腺癌中的表达、与患者分期及预后关系;RT-qPCR检测不同浓度瘦素处理后ACSL5的表达;慢病毒转染构建过表达ACSL5稳转株;Western blot检测上皮-间质转化(epithelial-mesenchymal transition,EMT)相关蛋白表达及AMPK-α、p-AMPK-α的表达。结果瘦素促进MCF-7细胞体外迁移和侵袭及EMT;ACSL5是瘦素下游靶基因,在乳腺癌中显著低表达且与患者预后相关;瘦素通过其受体OBR下调ACSL5的表达;瘦素激活AMPK通路下调ACSL5促进乳腺癌MCF-7细胞迁移和侵袭及EMT。结论瘦素可能通过激活AMPK通路下调ACSL5,增强乳腺癌细胞的迁移和侵袭能力并促进其EMT进程,从而促进乳腺癌恶性进展。
