丙型肝炎病毒(HCV)感染呈全球分布[1],而我国是丙型肝炎的高发区[2-4].HCV感染者80%发展成为慢性肝炎[5-7],而且发生肝硬变和肝细胞癌的危险性显著提高[8,9].最近证实低密度脂蛋白受 (low density lipoprotein receptor, LDL-R)可以识...丙型肝炎病毒(HCV)感染呈全球分布[1],而我国是丙型肝炎的高发区[2-4].HCV感染者80%发展成为慢性肝炎[5-7],而且发生肝硬变和肝细胞癌的危险性显著提高[8,9].最近证实低密度脂蛋白受 (low density lipoprotein receptor, LDL-R)可以识别和内化与低密度脂蛋白或极低密度脂蛋白结合HCV[10-12],从而确定LDL-R为HCV受体[13-15].LDL-R主要分布于肝实质细胞,能够结合并内化低密度脂蛋白(low density lipoprotein, LDL )和极低密度脂蛋白(very low density lipoprotein, VLDL),利用其中的胆固醇,从而参与胆固醇代谢,并调节血液胆固醇水平[16,17].展开更多
Objective:Clerodendrum glandulosum is widely used in traditional Chinese and Indian systems of medicine for conditions like hypertension and diabetes.While various pharmacological benefits have been reported,research ...Objective:Clerodendrum glandulosum is widely used in traditional Chinese and Indian systems of medicine for conditions like hypertension and diabetes.While various pharmacological benefits have been reported,research on its anti-atherosclerotic properties remains limited.Atherosclerosis(AS)is a chronic cardiovascular disease linked to dyslipidemia(DLD)and inflammation.This study aims to identify the bioactive fraction from C.glandulosum extract,evaluate its potential against AS and DLD,and explore the molecular mechanisms of cholesterol metabolism.Methods:Bioactivity-guided fractionation was employed to investigate the bioactivity of C.glandulosum by screening biochemical enzyme inhibitory potential.The active fraction was subjected to in vitro testing to assess the anti-inflammatory and anti-adhesion properties.The fraction was administered at 50and 100 mg/kg per os(p.o.)to cholesterol-cholic acid-thiouracil(CCT)diet-induced atherosclerotic Wistar rats.Changes in lipid and antioxidant profiles,inflammatory markers,and cholesterol metabolism pathways were assessed using Western blotting.Histopathological analyses of the aorta,liver,heart,and kidneys were also conducted.Molecular docking was conducted for the verbascoside(VER)and the standard statin,atorvastatin(ATS),for their binding capabilities with the molecular targets considered in this study.Results:Bioactivity-guided fractionation and screening revealed that ethyl acetate fraction(EAF)contained VER as the principal phytoconstituent.EAF exhibited potent enzyme inhibitory activity,with IC_(50)values of 1.059 mg/mL for pancreatic lipase and 22.48μg/mL for a-glucosidase.In vitro analysis revealed that EAF significantly lowered cell-to-cell adhesion to 0.57 folds from 2.5 folds in the disease control and normalized the inflammatory cytokines.In CCT-diet-induced rats,elevated serum cholesterol and low-density lipoproteins(LDL)levels(92.1 mg/dL and 78.49 mg/dL,respectively)were reduced to 63.52 mg/dL and 58.51 mg/dL with EAF at 100 mg/kg.EAF at 100 mg/kg reduced oxidized LDL to53.63 ng/mL compared to 157.1 ng/mL in CCT-diet-fed rats.EAF also restored antioxidant activity by increasing superoxide dismutase and catalase levels to 73.78 and 17.72 U/mg protein,respectively,compared to 42.22 and 9.62 U/mg protein in CCT-diet-fed rats.EAF restored inflammatory cytokines to normal levels.Histological analyses validated the protective benefits of EAF supplementation for the structural integrity of the aorta,liver,heart,and kidney tissues.Western blotting analysis of liver tissues revealed changes in the cholesterol metabolic pathway by upregulating peroxisome proliferatoractivated receptor gamma(PPARγ)/liver X receptor alpha(LXRa)/adenosine triphosphate-binding cassette sub-family G member 1(ABCG1)and low-density lipoprotein receptor(LDL-R)expression.Unlike ATS,molecular docking analyses indicated strong interactions between VER and molecular targets.Conclusion:EAF prevented DLD and AS by reverse cholesterol transport via the PPARγ/LXRa/ABCG1 pathway,offering potential therapeutic benefits for cardiovascular health.展开更多
目的建立豚鼠高胆固醇血症模型,并对形成机制进行探讨。方法高脂饲料诱导法建立豚鼠高胆固醇血症模型,分别于造模1、2、4周检测血清脂质的动态变化;检测肝脏脂质;酶联免疫分析法测定血清ox-LDL浓度;实时荧光定量PCR法检测肝脏CYP7A1(cho...目的建立豚鼠高胆固醇血症模型,并对形成机制进行探讨。方法高脂饲料诱导法建立豚鼠高胆固醇血症模型,分别于造模1、2、4周检测血清脂质的动态变化;检测肝脏脂质;酶联免疫分析法测定血清ox-LDL浓度;实时荧光定量PCR法检测肝脏CYP7A1(cholesterol 7a-hydroxylaseA1)、肝脏法尼酯X受体(hepatic farnesoid X receptor,FXR)、肝X受体α(liver X receptor,LXRα)、HMG-CoA还原酶mRNA的相对表达;Western blot法检测肝脏LDL-R的蛋白表达情况。结果豚鼠经高脂饲料诱导1周后,模型组与对照组比较血清TC、LDL-C即发生升高,随造模时间延长,血脂一直维持在较高水平。造模4周后模型组血清ox-LDL、sd-LDL浓度,肝脏TC水平比正常组升高。肝脏HMG-CoA还原酶表达下调,LDL-R蛋白表达量明显高于正常组。值得注意的是豚鼠肝脏法尼酯X受体(FXR)表达明显上调,且肝X受体α(LXRα)表达也明显上调,但二者激活水平相当,最终未改变肝脏CYP7A1mRNA的表达水平。结论高脂诱导豚鼠形成高胆固醇血症主要与外源性胆固醇和低密度脂蛋白的清除发生障碍有关。展开更多
文摘丙型肝炎病毒(HCV)感染呈全球分布[1],而我国是丙型肝炎的高发区[2-4].HCV感染者80%发展成为慢性肝炎[5-7],而且发生肝硬变和肝细胞癌的危险性显著提高[8,9].最近证实低密度脂蛋白受 (low density lipoprotein receptor, LDL-R)可以识别和内化与低密度脂蛋白或极低密度脂蛋白结合HCV[10-12],从而确定LDL-R为HCV受体[13-15].LDL-R主要分布于肝实质细胞,能够结合并内化低密度脂蛋白(low density lipoprotein, LDL )和极低密度脂蛋白(very low density lipoprotein, VLDL),利用其中的胆固醇,从而参与胆固醇代谢,并调节血液胆固醇水平[16,17].
基金supported by the In-house core-funded research project(No.IASST/R&D/ICP/IHP-19/2023-24/1301-1310)of the Institute of Advanced Study in Science and Technologythe Indian Council of Medical Research for the ICMR-SRF Fellowship granted to Ms.Puspanjali Khound(No.45/04/2022/TRM/BMS)+2 种基金Sophisticated Analytical Instrument CentreInstitute of Advanced Study in Science and Technologythe Department of Science and Technology,Government of India。
文摘Objective:Clerodendrum glandulosum is widely used in traditional Chinese and Indian systems of medicine for conditions like hypertension and diabetes.While various pharmacological benefits have been reported,research on its anti-atherosclerotic properties remains limited.Atherosclerosis(AS)is a chronic cardiovascular disease linked to dyslipidemia(DLD)and inflammation.This study aims to identify the bioactive fraction from C.glandulosum extract,evaluate its potential against AS and DLD,and explore the molecular mechanisms of cholesterol metabolism.Methods:Bioactivity-guided fractionation was employed to investigate the bioactivity of C.glandulosum by screening biochemical enzyme inhibitory potential.The active fraction was subjected to in vitro testing to assess the anti-inflammatory and anti-adhesion properties.The fraction was administered at 50and 100 mg/kg per os(p.o.)to cholesterol-cholic acid-thiouracil(CCT)diet-induced atherosclerotic Wistar rats.Changes in lipid and antioxidant profiles,inflammatory markers,and cholesterol metabolism pathways were assessed using Western blotting.Histopathological analyses of the aorta,liver,heart,and kidneys were also conducted.Molecular docking was conducted for the verbascoside(VER)and the standard statin,atorvastatin(ATS),for their binding capabilities with the molecular targets considered in this study.Results:Bioactivity-guided fractionation and screening revealed that ethyl acetate fraction(EAF)contained VER as the principal phytoconstituent.EAF exhibited potent enzyme inhibitory activity,with IC_(50)values of 1.059 mg/mL for pancreatic lipase and 22.48μg/mL for a-glucosidase.In vitro analysis revealed that EAF significantly lowered cell-to-cell adhesion to 0.57 folds from 2.5 folds in the disease control and normalized the inflammatory cytokines.In CCT-diet-induced rats,elevated serum cholesterol and low-density lipoproteins(LDL)levels(92.1 mg/dL and 78.49 mg/dL,respectively)were reduced to 63.52 mg/dL and 58.51 mg/dL with EAF at 100 mg/kg.EAF at 100 mg/kg reduced oxidized LDL to53.63 ng/mL compared to 157.1 ng/mL in CCT-diet-fed rats.EAF also restored antioxidant activity by increasing superoxide dismutase and catalase levels to 73.78 and 17.72 U/mg protein,respectively,compared to 42.22 and 9.62 U/mg protein in CCT-diet-fed rats.EAF restored inflammatory cytokines to normal levels.Histological analyses validated the protective benefits of EAF supplementation for the structural integrity of the aorta,liver,heart,and kidney tissues.Western blotting analysis of liver tissues revealed changes in the cholesterol metabolic pathway by upregulating peroxisome proliferatoractivated receptor gamma(PPARγ)/liver X receptor alpha(LXRa)/adenosine triphosphate-binding cassette sub-family G member 1(ABCG1)and low-density lipoprotein receptor(LDL-R)expression.Unlike ATS,molecular docking analyses indicated strong interactions between VER and molecular targets.Conclusion:EAF prevented DLD and AS by reverse cholesterol transport via the PPARγ/LXRa/ABCG1 pathway,offering potential therapeutic benefits for cardiovascular health.
文摘目的建立豚鼠高胆固醇血症模型,并对形成机制进行探讨。方法高脂饲料诱导法建立豚鼠高胆固醇血症模型,分别于造模1、2、4周检测血清脂质的动态变化;检测肝脏脂质;酶联免疫分析法测定血清ox-LDL浓度;实时荧光定量PCR法检测肝脏CYP7A1(cholesterol 7a-hydroxylaseA1)、肝脏法尼酯X受体(hepatic farnesoid X receptor,FXR)、肝X受体α(liver X receptor,LXRα)、HMG-CoA还原酶mRNA的相对表达;Western blot法检测肝脏LDL-R的蛋白表达情况。结果豚鼠经高脂饲料诱导1周后,模型组与对照组比较血清TC、LDL-C即发生升高,随造模时间延长,血脂一直维持在较高水平。造模4周后模型组血清ox-LDL、sd-LDL浓度,肝脏TC水平比正常组升高。肝脏HMG-CoA还原酶表达下调,LDL-R蛋白表达量明显高于正常组。值得注意的是豚鼠肝脏法尼酯X受体(FXR)表达明显上调,且肝X受体α(LXRα)表达也明显上调,但二者激活水平相当,最终未改变肝脏CYP7A1mRNA的表达水平。结论高脂诱导豚鼠形成高胆固醇血症主要与外源性胆固醇和低密度脂蛋白的清除发生障碍有关。
