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
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.
文摘目的探究糖尿病肾病(diabetic kidney disease,DKD)血瘀证大鼠肾损害与肾脏铁死亡的潜在机制。方法将50只SPF级雄性SD大鼠分为对照组、DKD组、DKD血瘀证组。采用腹腔注射链脲佐菌素的方法复制DKD大鼠模型,采用尾静脉注射右旋糖酐的方法复制DKD血瘀证模型。实验过程中观察大鼠血瘀证表现及检测24 h尿蛋白、血清肌酐、血尿素氮、血液流变学指标,采用苏木精-伊红染色、Masson染色、PAS染色观察肾脏的组织形态,采用透射电子显微镜观察铁死亡典型细胞的线粒体变化;采用免疫组织化学法、Western blot法检测肾组织铁死亡相关蛋白[长链酯酰辅酶A合成酶4(Acyl-CoA synthetase long chain family member 4,ACSL4)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)]及肾脏纤维化指标[纤维连接蛋白(fibronectin,FN)、Ⅳ型胶原蛋白(typeⅣcollagen,Col-Ⅳ)]的表达水平。结果与对照组比较,DKD组大鼠肾脏病理变化加重,线粒体损伤明显,24 h尿蛋白含量,血清肌酐、血尿素氮水平及全血黏度、血浆黏度明显升高(P<0.05),肾脏ACSL4、FN和Col-Ⅳ及其mRNA表达水平明显升高(P<0.05),GPX4蛋白及其mRNA表达水平明显降低(P<0.05)。DKD血瘀证大鼠出现唇色黯淡、眼球黯红、耳廓紫红、舌下脉络紫黯等血瘀证表现,24 h尿蛋白和血清肌酐、血尿素氮水平明显高于DKD组,光学显微镜下可见肾脏出现明显的系膜基质增生、肾小球萎缩、肾间质胶原沉积和纤维化,电子显微镜下可见肾组织细胞线粒体损伤明显,嵴基本断裂,ACSL4、FN、Col-Ⅳ及其mRNA表达水平较DKD组明显上升(P<0.05),GPX4蛋白及其mRNA表达水平较DKD组明显下降(P<0.05)。结论DKD血瘀证大鼠肾脏损害更加严重,其机制可能与GPX4、ACSL4介导的铁死亡有关。
