PDF(Pubmed)
Abstract：
UNASSIGNED: The role of platelet autophagy in cirrhotic thrombocytopenia (CTP) remains unclear. This study aimed to investigate the impact of platelet autophagy in CTP and elucidate the regulatory mechanism of hydrogen sulfide (H2S) on platelet autophagy.
UNASSIGNED: Platelets from 56 cirrhotic patients and 56 healthy individuals were isolated for in vitro analyses. Autophagy markers (ATG7, BECN1, LC3, and SQSTM1) were quantified using enzyme-linked immunosorbent assay, while autophagosomes were visualized through electron microscopy. Western blotting was used to assess the autophagy-related proteins and the PDGFR/PI3K/Akt/mTOR pathway following treatment with NaHS (an H2S donor), hydroxocobalamin (an H2S scavenger), or AG 1295 (a selective PDGFR-α inhibitor). A carbon tetrachloride-induced cirrhotic BALB/c mouse model was established. Cirrhotic mice with thrombocytopenia were randomly treated with normal saline, NaHS, or hydroxocobalamin for 15 days. Changes in platelet count and aggregation rate were observed every three days.
UNASSIGNED: Cirrhotic patients with thrombocytopenia exhibited significantly decreased platelet autophagy markers and endogenous H2S levels, alongside increased platelet aggregation, compared to healthy controls. In vitro, NaHS treatment of platelets from severe CTP patients elevated LC3-II levels, reduced SQSTM1 levels, and decreased platelet aggregation in a dose-dependent manner. H2S treatment inhibited PDGFR, PI3K, Akt, and mTOR phosphorylation. In vivo, NaHS significantly increased LC3-II and decreased SQSTM1 expressions in platelets of cirrhotic mice, reducing platelet aggregation without affecting the platelet count.
UNASSIGNED: Diminished platelet autophagy potentially contributes to thrombocytopenia in cirrhotic patients. H2S modulates platelet autophagy and functions possibly via the PDGFR-α/PI3K/Akt/mTOR signaling pathway.
UNASSIGNED: Platelets from 56 cirrhotic patients and 56 healthy individuals were isolated for in vitro analyses. Autophagy markers (ATG7, BECN1, LC3, and SQSTM1) were quantified using enzyme-linked immunosorbent assay, while autophagosomes were visualized through electron microscopy. Western blotting was used to assess the autophagy-related proteins and the PDGFR/PI3K/Akt/mTOR pathway following treatment with NaHS (an H2S donor), hydroxocobalamin (an H2S scavenger), or AG 1295 (a selective PDGFR-α inhibitor). A carbon tetrachloride-induced cirrhotic BALB/c mouse model was established. Cirrhotic mice with thrombocytopenia were randomly treated with normal saline, NaHS, or hydroxocobalamin for 15 days. Changes in platelet count and aggregation rate were observed every three days.
UNASSIGNED: Cirrhotic patients with thrombocytopenia exhibited significantly decreased platelet autophagy markers and endogenous H2S levels, alongside increased platelet aggregation, compared to healthy controls. In vitro, NaHS treatment of platelets from severe CTP patients elevated LC3-II levels, reduced SQSTM1 levels, and decreased platelet aggregation in a dose-dependent manner. H2S treatment inhibited PDGFR, PI3K, Akt, and mTOR phosphorylation. In vivo, NaHS significantly increased LC3-II and decreased SQSTM1 expressions in platelets of cirrhotic mice, reducing platelet aggregation without affecting the platelet count.
UNASSIGNED: Diminished platelet autophagy potentially contributes to thrombocytopenia in cirrhotic patients. H2S modulates platelet autophagy and functions possibly via the PDGFR-α/PI3K/Akt/mTOR signaling pathway.
摘要：
■血小板自噬在肝硬化血小板减少症（CTP）中的作用尚不清楚。本研究旨在研究血小板自噬在CTP中的作用,阐明硫化氢(H2S)对血小板自噬的调控机制。
■分离来自56名肝硬化患者和56名健康个体的血小板用于体外分析。自噬标志物（ATG7，BECN1，LC3和SQSTM1）使用酶联免疫吸附试验进行定量，而自噬体通过电子显微镜观察。Western印迹用于评估NaHS（H2S供体）治疗后的自噬相关蛋白和PDGFR/PI3K/Akt/mTOR途径，羟钴胺（H2S清除剂），或AG1295（选择性PDGFR-α抑制剂）。建立四氯化碳诱导的肝硬化BALB/c小鼠模型。肝硬化小鼠血小板减少症随机给予生理盐水,NaHS,或羟钴胺15天。每三天观察血小板计数和聚集率的变化。
■伴血小板减少症的肝硬化患者表现出显著降低的血小板自噬标志物和内源性H2S水平，同时增加血小板聚集，与健康对照相比。体外，NaHS治疗严重CTP患者血小板LC3-II水平升高，降低SQSTM1级别，并以剂量依赖性方式降低血小板聚集。H2S处理抑制PDGFR，PI3K,Akt,和mTOR磷酸化。在体内,NaHS显著增加肝硬化小鼠血小板中LC3-II和SQSTM1的表达,减少血小板聚集而不影响血小板计数。
■血小板自噬减少可能导致肝硬化患者的血小板减少。H2S调节血小板自噬并可能通过PDGFR-α/PI3K/Akt/mTOR信号通路发挥作用。
■分离来自56名肝硬化患者和56名健康个体的血小板用于体外分析。自噬标志物（ATG7，BECN1，LC3和SQSTM1）使用酶联免疫吸附试验进行定量，而自噬体通过电子显微镜观察。Western印迹用于评估NaHS（H2S供体）治疗后的自噬相关蛋白和PDGFR/PI3K/Akt/mTOR途径，羟钴胺（H2S清除剂），或AG1295（选择性PDGFR-α抑制剂）。建立四氯化碳诱导的肝硬化BALB/c小鼠模型。肝硬化小鼠血小板减少症随机给予生理盐水,NaHS,或羟钴胺15天。每三天观察血小板计数和聚集率的变化。
■伴血小板减少症的肝硬化患者表现出显著降低的血小板自噬标志物和内源性H2S水平，同时增加血小板聚集，与健康对照相比。体外，NaHS治疗严重CTP患者血小板LC3-II水平升高，降低SQSTM1级别，并以剂量依赖性方式降低血小板聚集。H2S处理抑制PDGFR，PI3K,Akt,和mTOR磷酸化。在体内,NaHS显著增加肝硬化小鼠血小板中LC3-II和SQSTM1的表达,减少血小板聚集而不影响血小板计数。
■血小板自噬减少可能导致肝硬化患者的血小板减少。H2S调节血小板自噬并可能通过PDGFR-α/PI3K/Akt/mTOR信号通路发挥作用。
