PDF(Pubmed)
Abstract：
BACKGROUND: Nonalcoholic steatohepatitis (NASH) is a prevalent chronic liver condition. However, the potential therapeutic benefits and underlying mechanism of nicotinate-curcumin (NC) in the treatment of NASH remain uncertain.
METHODS: A rat model of NASH induced by a high-fat and high-fructose diet was treated with nicotinate-curcumin (NC, 20, 40 mg·kg- 1), curcumin (Cur, 40 mg·kg- 1) and metformin (Met, 50 mg·kg- 1) for a duration of 4 weeks. The interaction between NASH, Cur and Aldo-Keto reductase family 1 member B10 (AKR1B10) was filter and analyzed using network pharmacology. The interaction of Cur, NC and AKR1B10 was analyzed using molecular docking techniques, and the binding energy of Cur and NC with AKR1B10 was compared. HepG2 cells were induced by Ox-LDL (25 µg·ml- 1, 24 h) in high glucose medium. NC (20µM, 40µM), Cur (40µM) Met (150µM) and epalrestat (Epa, 75µM) were administered individually. The activities of ALT, AST, ALP and the levels of LDL, HDL, TG, TC and FFA in serum were quantified using a chemiluminescence assay. Based on the changes in the above indicators, score according to NAS standards. The activities of Acetyl-CoA and Malonyl-CoA were measured using an ELISA assay. And the expression and cellular localization of AKR1B10 and Acetyl-CoA carboxylase (ACCα) in HepG2 cells were detected by Western blotting and immunofluorescence.
RESULTS: The results of the animal experiments demonstrated that NASH rat model induced by a high-fat and high-fructose diet exhibited pronounced dysfunction in liver function and lipid metabolism. Additionally, there was a significant increase in serum levels of FFA and TG, as well as elevated expression of AKR1B10 and ACCα, and heightened activity of Acetyl-CoA and Malonyl-CoA in liver tissue. The administration of NC showed to enhance liver function in rats with NASH, leading to reductions in ALT, AST and ALP levels, and decrease in blood lipid and significant inhibition of FFA and TG synthesis in the liver. Network pharmacological analysis identified AKR1B10 and ACCα as potential targets for NASH treatment. Molecular docking studies revealed that both Cur and NC are capable of binding to AKR1B10, with NC exhibiting a stronger binding energy to AKR1B10. Western blot analysis demonstrated an upregulation in the expression of AKR1B10 and ACCα in the liver tissue of NASH rats, accompanied by elevated Acetyl-CoA and Malonyl-CoA activity, and increased levels of FFA and TG. The results of the HepG2 cell experiments induced by Ox-LDL suggest that NC significantly inhibited the expression and co-localization of AKR1B10 and ACCα, while also reduced levels of TC and LDL-C and increased level of HDL-C. These effects are accompanied by a decrease in the activities of ACCα and Malonyl-CoA, and levels of FFA and TG. Furthermore, the impact of NC appears to be more pronounced compared to Cur.
CONCLUSIONS: NC could effectively treat NASH and improve liver function and lipid metabolism disorder. The mechanism of NC is related to the inhibition of AKR1B10/ACCα pathway and FFA/TG synthesis of liver.
METHODS: A rat model of NASH induced by a high-fat and high-fructose diet was treated with nicotinate-curcumin (NC, 20, 40 mg·kg- 1), curcumin (Cur, 40 mg·kg- 1) and metformin (Met, 50 mg·kg- 1) for a duration of 4 weeks. The interaction between NASH, Cur and Aldo-Keto reductase family 1 member B10 (AKR1B10) was filter and analyzed using network pharmacology. The interaction of Cur, NC and AKR1B10 was analyzed using molecular docking techniques, and the binding energy of Cur and NC with AKR1B10 was compared. HepG2 cells were induced by Ox-LDL (25 µg·ml- 1, 24 h) in high glucose medium. NC (20µM, 40µM), Cur (40µM) Met (150µM) and epalrestat (Epa, 75µM) were administered individually. The activities of ALT, AST, ALP and the levels of LDL, HDL, TG, TC and FFA in serum were quantified using a chemiluminescence assay. Based on the changes in the above indicators, score according to NAS standards. The activities of Acetyl-CoA and Malonyl-CoA were measured using an ELISA assay. And the expression and cellular localization of AKR1B10 and Acetyl-CoA carboxylase (ACCα) in HepG2 cells were detected by Western blotting and immunofluorescence.
RESULTS: The results of the animal experiments demonstrated that NASH rat model induced by a high-fat and high-fructose diet exhibited pronounced dysfunction in liver function and lipid metabolism. Additionally, there was a significant increase in serum levels of FFA and TG, as well as elevated expression of AKR1B10 and ACCα, and heightened activity of Acetyl-CoA and Malonyl-CoA in liver tissue. The administration of NC showed to enhance liver function in rats with NASH, leading to reductions in ALT, AST and ALP levels, and decrease in blood lipid and significant inhibition of FFA and TG synthesis in the liver. Network pharmacological analysis identified AKR1B10 and ACCα as potential targets for NASH treatment. Molecular docking studies revealed that both Cur and NC are capable of binding to AKR1B10, with NC exhibiting a stronger binding energy to AKR1B10. Western blot analysis demonstrated an upregulation in the expression of AKR1B10 and ACCα in the liver tissue of NASH rats, accompanied by elevated Acetyl-CoA and Malonyl-CoA activity, and increased levels of FFA and TG. The results of the HepG2 cell experiments induced by Ox-LDL suggest that NC significantly inhibited the expression and co-localization of AKR1B10 and ACCα, while also reduced levels of TC and LDL-C and increased level of HDL-C. These effects are accompanied by a decrease in the activities of ACCα and Malonyl-CoA, and levels of FFA and TG. Furthermore, the impact of NC appears to be more pronounced compared to Cur.
CONCLUSIONS: NC could effectively treat NASH and improve liver function and lipid metabolism disorder. The mechanism of NC is related to the inhibition of AKR1B10/ACCα pathway and FFA/TG synthesis of liver.
摘要：
背景：非酒精性脂肪性肝炎（NASH）是一种常见的慢性肝病。然而,烟酸-姜黄素(NC)治疗NASH的潜在疗效和潜在机制尚不确定.
方法：用烟酸-姜黄素治疗高脂高果糖饮食诱导的NASH大鼠模型（NC，20,40mg·kg-1),姜黄素（Cur，40mg·kg-1)和二甲双胍(Met，50mg·kg-1），持续4周。NASH之间的相互作用，使用网络药理学对Cur和Aldo-Keto还原酶家族1成员B10(AKR1B10)进行过滤和分析。库尔的互动，使用分子对接技术分析了NC和AKR1B10，并比较了Cur和NC与AKR1B10的结合能。在高葡萄糖培养基中，Ox-LDL（25µg·ml-1，24h）诱导HepG2细胞。NC(20µM,40µM),Cur(40µM)Met(150µM)andepalrestat(Epa,75µM）分别给药。ALT的活动，AST,ALP和LDL水平，HDL,TG,使用化学发光测定法定量血清中的TC和FFA。根据上述指标的变化，根据NAS标准进行评分。使用ELISA测定法测量乙酰辅酶A和丙二酰辅酶A的活性。采用免疫印迹法和免疫荧光法检测HepG2细胞中AKR1B10和乙酰辅酶A羧化酶(ACCα)的表达和细胞定位。
结果：动物实验结果表明，高脂高果糖饮食诱导的NASH大鼠模型表现出明显的肝功能和脂质代谢障碍。此外,血清FFA和TG水平显着升高，以及AKR1B10和ACCα的表达升高，肝组织中乙酰辅酶A和丙二酰辅酶A的活性增强。给药NC可增强NASH大鼠的肝功能，导致ALT减少，AST和ALP水平，并降低血脂，并显着抑制肝脏中的FFA和TG合成。网络药理学分析确定AKR1B10和ACCα为NASH治疗的潜在靶标。分子对接研究表明，Cur和NC都能够与AKR1B10结合，其中NC对AKR1B10表现出更强的结合能。Westernblot分析显示NASH大鼠肝组织中AKR1B10和ACCα的表达上调,伴有乙酰辅酶A和丙二酰辅酶A活性升高，和增加的FFA和TG水平。Ox-LDL诱导的HepG2细胞实验结果表明，NC显著抑制AKR1B10和ACCα的表达和共定位，同时也降低了TC和LDL-C的水平和升高的HDL-C的水平。这些作用伴随着ACCα和丙二酰辅酶A活性的降低，以及FFA和TG的水平。此外，与Cur相比，NC的影响似乎更明显。
结论：NC能有效治疗NASH，改善肝功能和脂代谢紊乱。其机制与抑制肝脏AKR1B10/ACCα通路和FFA/TG合成有关。
方法：用烟酸-姜黄素治疗高脂高果糖饮食诱导的NASH大鼠模型（NC，20,40mg·kg-1),姜黄素（Cur，40mg·kg-1)和二甲双胍(Met，50mg·kg-1），持续4周。NASH之间的相互作用，使用网络药理学对Cur和Aldo-Keto还原酶家族1成员B10(AKR1B10)进行过滤和分析。库尔的互动，使用分子对接技术分析了NC和AKR1B10，并比较了Cur和NC与AKR1B10的结合能。在高葡萄糖培养基中，Ox-LDL（25µg·ml-1，24h）诱导HepG2细胞。NC(20µM,40µM),Cur(40µM)Met(150µM)andepalrestat(Epa,75µM）分别给药。ALT的活动，AST,ALP和LDL水平，HDL,TG,使用化学发光测定法定量血清中的TC和FFA。根据上述指标的变化，根据NAS标准进行评分。使用ELISA测定法测量乙酰辅酶A和丙二酰辅酶A的活性。采用免疫印迹法和免疫荧光法检测HepG2细胞中AKR1B10和乙酰辅酶A羧化酶(ACCα)的表达和细胞定位。
结果：动物实验结果表明，高脂高果糖饮食诱导的NASH大鼠模型表现出明显的肝功能和脂质代谢障碍。此外,血清FFA和TG水平显着升高，以及AKR1B10和ACCα的表达升高，肝组织中乙酰辅酶A和丙二酰辅酶A的活性增强。给药NC可增强NASH大鼠的肝功能，导致ALT减少，AST和ALP水平，并降低血脂，并显着抑制肝脏中的FFA和TG合成。网络药理学分析确定AKR1B10和ACCα为NASH治疗的潜在靶标。分子对接研究表明，Cur和NC都能够与AKR1B10结合，其中NC对AKR1B10表现出更强的结合能。Westernblot分析显示NASH大鼠肝组织中AKR1B10和ACCα的表达上调,伴有乙酰辅酶A和丙二酰辅酶A活性升高，和增加的FFA和TG水平。Ox-LDL诱导的HepG2细胞实验结果表明，NC显著抑制AKR1B10和ACCα的表达和共定位，同时也降低了TC和LDL-C的水平和升高的HDL-C的水平。这些作用伴随着ACCα和丙二酰辅酶A活性的降低，以及FFA和TG的水平。此外，与Cur相比，NC的影响似乎更明显。
结论：NC能有效治疗NASH，改善肝功能和脂代谢紊乱。其机制与抑制肝脏AKR1B10/ACCα通路和FFA/TG合成有关。
