AIM To evaluate the levels of mi R-192-5 p in non-alcoholic fatty liver disease(NAFLD) models and demonstrate the role of mi R-192-5 p in lipid accumulation. METHODS Thirty Sprague Dawley rats were randomly divided in...AIM To evaluate the levels of mi R-192-5 p in non-alcoholic fatty liver disease(NAFLD) models and demonstrate the role of mi R-192-5 p in lipid accumulation. METHODS Thirty Sprague Dawley rats were randomly divided into three groups, which were given a standard diet, a high-fat diet(HFD), and an HFD with injection of liraglutide. At the end of 16 weeks, hepatic mi R-192-5 p and stearoyl-Co A desaturase 1(SCD-1) levels were measured. Mi R-192-5 p mimic and inhibitor and SCD-1 si RNA were transfected into Huh7 cells exposed to palmitic acid(PA). Lipid accumulation was evaluated by oil red O staining and triglyceride assays. Direct interaction was validated by dual-luciferase reporter gene assays.RESULTS The HFD rats showed a 0.46-fold decrease and a 3.5-fold increase in hepatic mi R-192-5 p and SCD-1 protein levels compared with controls, respectively, which could be reversed after disease remission by liraglutide injection(P < 0.01). The Huh7 cells exposed to PA also showed down-regulation and up-regulation of mi R-192-5 p and SCD-1 protein levels, respectively(P < 0.01). Transfection with mi R-192-5 p mimic and inhibitor in Huh7 cells induced dramatic repression and promotion of SCD-1 protein levels, respectively(P < 0.01). Luciferase activity was suppressed and enhanced by mi R-192-5 p mimic and inhibitor, respectively, in wild-type SCD-1(P < 0.01) but not in mutant SCD-1. Mi R-192-5 p overexpression reduced lipid accumulation significantly in PA-treated Huh7 cells, and SCD-1 si RNA transfection abrogated the lipid deposition aggravated by mi R-192-5 p inhibitor(P < 0.01).CONCLUSION This study demonstrates that mi R-192-5 p has a negative regulatory role in lipid synthesis, which is mediated through its direct regulation of SCD-1.展开更多
AIM To compare liver proteolysis and proteasome activation in steatotic liver grafts conserved in University of Wisconsin(UW) and Institut Georges Lopez-1(IGL-1)solutions.METHODS Fatty liver grafts from male obese Z&#...AIM To compare liver proteolysis and proteasome activation in steatotic liver grafts conserved in University of Wisconsin(UW) and Institut Georges Lopez-1(IGL-1)solutions.METHODS Fatty liver grafts from male obese Zücker rats were conserved in UW and IGL-1 solutions for 24 h at 4 ℃and subjected to "ex vivo " normo-thermic perfusion(2 h; 37 ℃). Liver proteolysis in tissue specimens and perfusate was measured by reverse-phase high performance liquid chromatography. Total free amino acid release was correlated with the activation of the ubiquitin proteasome system(UPS: measured as chymotryptic-like activity and 20 S and 19 S proteasome), the prevention of liver injury(transaminases), mitochondrial injury(confocal microscopy) and inflammation markers(TNF 1 alpha, high mobility group box-1(HGMB-1) and PPAR gamma), and liver apoptosis(TUNEL assay, cytochrome c and caspase 3).RESULTS Profiles of free AA(alanine, proline, leucine, isoleucine, methionine, lysine, ornithine, and threonine, among others) were similar for tissue and reperfusion effluent. In all cases, the IGL-1 solution showed a significantly higher prevention of proteolysis than UW(P < 0.05) after cold ischemia reperfusion. Livers conserved in IGL-1 presented more effective prevention of ATP-breakdown and more inhibition of UPS activity(measured as chymotryptic-like activity). In addition, the prevention of liver proteolysis and UPS activation correlated with the prevention of liver injury(AST/ALT) and mitochondrial damage(revealed by confocal microscopy findings) as well as with the prevention of inflammatory markers(TNF1alpha and HMGB) after reperfusion. In addition, the liver grafts preserved in IGL-1 showed a significant decrease in liver apoptosis, as shown by TUNEL assay and the reduction of cytochrome c, caspase 3 and P62 levels. CONCLUSION Our comparison of these two preservation solutions suggests that IGL-1 helps to prevent ATP breakdown more effectively than UW and subsequently achieves a higher UPS inhibition and reduced liver proteolysis.展开更多
目的 探讨非酒精性脂肪肝(NAFLD)患者血清高迁移率族蛋白B1(HMGB1)、白细胞介素-1β(IL-1β)、脂肪特异性丝氨酸蛋白酶抑制剂(Vaspin)水平与胰岛素抵抗指数(HOMA-IR)的相关性。方法 选取2016年7月至2018年10月该院收治的188例NAFLD患者...目的 探讨非酒精性脂肪肝(NAFLD)患者血清高迁移率族蛋白B1(HMGB1)、白细胞介素-1β(IL-1β)、脂肪特异性丝氨酸蛋白酶抑制剂(Vaspin)水平与胰岛素抵抗指数(HOMA-IR)的相关性。方法 选取2016年7月至2018年10月该院收治的188例NAFLD患者作为研究对象。选取同期进行体检的健康者100例作为健康对照组。比较2组间的基本资料、HMGB1、IL-1β、Vaspin水平,进行单因素分析、多元Logistic回归分析及Pearson相关分析。结果 与健康对照组比较,NAFLD组总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、空腹血糖(FPG)、空腹胰岛素(FINS)、HOMA-IR、HMGB-1、IL-1β、Vaspin均显著高于健康对照组( P <0.05)。多元Logistic回归分析结果显示,HMGB-1、IL-1β、Vaspin是NAFLD患者胰岛素抵抗的独立影响因素。随着患者病情严重程度的加重,HMGB-1、IL-1β、Vaspin、FINS、HOMA-IR水平不断升高( P <0.05)。Pearson相关性分析结果显示,HMGB-1、IL-1β、Vaspin与NAFLD患者FINS、HOMA-IR均呈正相关( P <0.05)。结论 HMGB-1、IL-1β、Vaspin可能参与NAFLD患者胰岛素抵抗现象的发生、发展。展开更多
目的研究不同类型脂肪肝患者血清高迁移率族蛋白B1(high mobility group box 1,HMGB1)水平、氧化应激指标及炎性因子水平,对其与脂肪肝的关系及临床意义进行了探讨。方法脂肪肝患者640例中酒精性脂肪肝(AFLD)组270例,非酒精性脂肪肝(NAF...目的研究不同类型脂肪肝患者血清高迁移率族蛋白B1(high mobility group box 1,HMGB1)水平、氧化应激指标及炎性因子水平,对其与脂肪肝的关系及临床意义进行了探讨。方法脂肪肝患者640例中酒精性脂肪肝(AFLD)组270例,非酒精性脂肪肝(NAFLD)组370例;以健康体检者300例为对照组。应用ELISA技术检测脂肪肝患者血清中HMGB1水平;同时检测氧化应激指标丙二醛(MDA)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)及炎性因子白介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)的水平。结果 NAFLD组、AFLD组的TC、TG、LDLC、ALT、AST、GLU的水平均明显高于对照组,HDLC的水平明显低于对照组,差异均有统计学意义(P<0.05),NAFLD组与AFLD组上述指标差异无统计学意义(P>0.05);NAFLD组BMI明显高于AFLD组与对照组,差异有统计学意义(P<0.05);NAFLD组、AFLD组血清MDA水平明显高于对照组,血清SOD、GSH水平明显低于对照组,差异均有统计学意义(P<0.05),以NAFLD组变化更明显。NAFLD组、AFLD组血清HMGB1、IL-6、TNF-α、IL-8水平明显高于对照组,差异有统计学意义(P<0.05),以NAFLD组升高更明显。NAFLD组、AFLD组外周血HMGB1水平与外周血IL-6、TNF-α、IL-8、MDA水平呈正相关性(r值分别为0.3976、0.4217、0.4981、0.3124、0.3275、0.3976),与外周血SOD、GSH水平呈负相关(r值分别为-0.3326、-0.3652,-0.2988、-0.3106,P<0.05)。结论不同类型脂肪肝患者血清HMGB1水平升高,HMGB1可能通过调控炎性因子及应激指标而参与脂肪肝的发生与进展。展开更多
基金Supported by National Key R&D Program of China No.2017YFC0908900National Key Basic Research Project,No.2012CB517501National Natural Science Foundation of China,No.81470840 and No.81600464
文摘AIM To evaluate the levels of mi R-192-5 p in non-alcoholic fatty liver disease(NAFLD) models and demonstrate the role of mi R-192-5 p in lipid accumulation. METHODS Thirty Sprague Dawley rats were randomly divided into three groups, which were given a standard diet, a high-fat diet(HFD), and an HFD with injection of liraglutide. At the end of 16 weeks, hepatic mi R-192-5 p and stearoyl-Co A desaturase 1(SCD-1) levels were measured. Mi R-192-5 p mimic and inhibitor and SCD-1 si RNA were transfected into Huh7 cells exposed to palmitic acid(PA). Lipid accumulation was evaluated by oil red O staining and triglyceride assays. Direct interaction was validated by dual-luciferase reporter gene assays.RESULTS The HFD rats showed a 0.46-fold decrease and a 3.5-fold increase in hepatic mi R-192-5 p and SCD-1 protein levels compared with controls, respectively, which could be reversed after disease remission by liraglutide injection(P < 0.01). The Huh7 cells exposed to PA also showed down-regulation and up-regulation of mi R-192-5 p and SCD-1 protein levels, respectively(P < 0.01). Transfection with mi R-192-5 p mimic and inhibitor in Huh7 cells induced dramatic repression and promotion of SCD-1 protein levels, respectively(P < 0.01). Luciferase activity was suppressed and enhanced by mi R-192-5 p mimic and inhibitor, respectively, in wild-type SCD-1(P < 0.01) but not in mutant SCD-1. Mi R-192-5 p overexpression reduced lipid accumulation significantly in PA-treated Huh7 cells, and SCD-1 si RNA transfection abrogated the lipid deposition aggravated by mi R-192-5 p inhibitor(P < 0.01).CONCLUSION This study demonstrates that mi R-192-5 p has a negative regulatory role in lipid synthesis, which is mediated through its direct regulation of SCD-1.
基金Supported by Instituto de Salud Carlos III(ISCIII)through the FIS project PI12/0056,co-funded by FEDER from Regional Development European Funds(European Union)
文摘AIM To compare liver proteolysis and proteasome activation in steatotic liver grafts conserved in University of Wisconsin(UW) and Institut Georges Lopez-1(IGL-1)solutions.METHODS Fatty liver grafts from male obese Zücker rats were conserved in UW and IGL-1 solutions for 24 h at 4 ℃and subjected to "ex vivo " normo-thermic perfusion(2 h; 37 ℃). Liver proteolysis in tissue specimens and perfusate was measured by reverse-phase high performance liquid chromatography. Total free amino acid release was correlated with the activation of the ubiquitin proteasome system(UPS: measured as chymotryptic-like activity and 20 S and 19 S proteasome), the prevention of liver injury(transaminases), mitochondrial injury(confocal microscopy) and inflammation markers(TNF 1 alpha, high mobility group box-1(HGMB-1) and PPAR gamma), and liver apoptosis(TUNEL assay, cytochrome c and caspase 3).RESULTS Profiles of free AA(alanine, proline, leucine, isoleucine, methionine, lysine, ornithine, and threonine, among others) were similar for tissue and reperfusion effluent. In all cases, the IGL-1 solution showed a significantly higher prevention of proteolysis than UW(P < 0.05) after cold ischemia reperfusion. Livers conserved in IGL-1 presented more effective prevention of ATP-breakdown and more inhibition of UPS activity(measured as chymotryptic-like activity). In addition, the prevention of liver proteolysis and UPS activation correlated with the prevention of liver injury(AST/ALT) and mitochondrial damage(revealed by confocal microscopy findings) as well as with the prevention of inflammatory markers(TNF1alpha and HMGB) after reperfusion. In addition, the liver grafts preserved in IGL-1 showed a significant decrease in liver apoptosis, as shown by TUNEL assay and the reduction of cytochrome c, caspase 3 and P62 levels. CONCLUSION Our comparison of these two preservation solutions suggests that IGL-1 helps to prevent ATP breakdown more effectively than UW and subsequently achieves a higher UPS inhibition and reduced liver proteolysis.
文摘目的 探讨非酒精性脂肪肝(NAFLD)患者血清高迁移率族蛋白B1(HMGB1)、白细胞介素-1β(IL-1β)、脂肪特异性丝氨酸蛋白酶抑制剂(Vaspin)水平与胰岛素抵抗指数(HOMA-IR)的相关性。方法 选取2016年7月至2018年10月该院收治的188例NAFLD患者作为研究对象。选取同期进行体检的健康者100例作为健康对照组。比较2组间的基本资料、HMGB1、IL-1β、Vaspin水平,进行单因素分析、多元Logistic回归分析及Pearson相关分析。结果 与健康对照组比较,NAFLD组总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、空腹血糖(FPG)、空腹胰岛素(FINS)、HOMA-IR、HMGB-1、IL-1β、Vaspin均显著高于健康对照组( P <0.05)。多元Logistic回归分析结果显示,HMGB-1、IL-1β、Vaspin是NAFLD患者胰岛素抵抗的独立影响因素。随着患者病情严重程度的加重,HMGB-1、IL-1β、Vaspin、FINS、HOMA-IR水平不断升高( P <0.05)。Pearson相关性分析结果显示,HMGB-1、IL-1β、Vaspin与NAFLD患者FINS、HOMA-IR均呈正相关( P <0.05)。结论 HMGB-1、IL-1β、Vaspin可能参与NAFLD患者胰岛素抵抗现象的发生、发展。
文摘目的:观察白芍总苷(TGP)对非酒精性脂肪性肝病(NAFLD)大鼠高迁移率族蛋白1(HMGB1)、晚期糖基化终产物受体(RAGE)通路的调控作用。方法:用高脂-高果糖餐诱导NAFLD大鼠模型,将模型大鼠随机分为模型组,TGP高、低剂量组(200、100 mg·kg^(-1)·d^(-1)),二甲双胍组(200 mg·kg^(-1)·d^(-1)),水飞蓟宾组(200 mg·kg^(-1)·d^(-1)),并另设正常对照组。用药6周后观察各组大鼠胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、甘油三酯(TG)、游离脂肪酸(FFA)、丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、空腹血糖(FBG)、餐后2 h血糖(2 h BG)、胰岛素(Fins)水平,计算胰岛素抵抗指数(HOMA-IR)和肝脏指数。Western blot方法检测肝组织中HMGB1、RAGE蛋白活性。结果:与模型组比较,TGP高、低剂量组LDL-C、TG、TC、FFA、ALT、AST、2 h BG、Fins、HOMA-IR均明显降低(P<0.01或P<0.05),高、低剂量TGP在拮抗胰岛素抵抗、降糖降脂、改善肝功能方面有较显著的作用,TGP高、低剂量组均可下调HMGB1(P<0.01)和RAGE(P<0.05)蛋白的表达。结论:TGP通过抑制HMGB1、RAGE信号通路的转导而起到改善NAFLD大鼠的糖脂代谢异常,拮抗胰岛素抵抗,改善肝功能的作用。
