Objective:This study aimed to elucidate the effect of the lipopolysaccharides/toll-like receptor 4(LPS/TLR4)pathway on early atherosclerosis(AS)development and its associated changes in fecal metabolites,thereby provi...Objective:This study aimed to elucidate the effect of the lipopolysaccharides/toll-like receptor 4(LPS/TLR4)pathway on early atherosclerosis(AS)development and its associated changes in fecal metabolites,thereby providing an experimental foundation for strategies to prevent and treat early AS.Methods:Twelve Tibetan miniature pigs aged 4-5 months were divided into normal control(NC)group and AS group(6 pigs in each).The group assignment was primarily based on body weight;Secondary criteria,including glucose,lipid profiles,and inflammatory indices,were considered to ensure balanced baseline characteristics between the 2 groups(all P>0.05).AS group received a high-fat diet for 16 weeks to establish an AS model,while the NC group received a normal diet.Subsequently,serum levels of lipids and various inflammation and oxidative stress markers were measured.Pathological changes in the aorta and colon tissue,LPS/TLR4 pathway-associated protein expressions in the aorta,as well as occludin and zonula occludens-1 in the colon were also assessed.Proton nuclear magnetic resonance spectra technology was employed for the metabolomic analysis of fecal extracts.Results:The lipid metabolism was disrupted in AS group,with significantly higher total cholesterol,high-density lipoprotein cholesterol,and low-density lipoprotein cholesterol levels((12.24±5.24)mmol/L vs.(1.86±0.27)mmol/L,P=0.004,6;(2.39±0.50)mmol/L vs.(0.83±0.07)mmol/L,P=0.000,5;(6.94±2.87)mmol/L vs.(0.77±0.18)mmol/L,P=0.003,3),as compared to that in NC group.Serum factors,including LPS,tumor necrosis factor-α,and malondialdehyde levels of AS group were significantly higher than that of NC group((1,230.00±192.70)EU/L vs.(695.70±213.70)EU/L),P=0.001,1;(424.20±176.90)ng/L vs.(51.20±26.61)ng/L,P=0.023,5;(3.60±0.77)nmol/mL vs.(2.62±0.21)nmol/mL,P=0.025,4).Pathological evaluations revealed prominent lipid deposition area in the aortic arch,thoracic aorta,and abdominal aorta of the AS group compared with that of the NC group(4.17%±2.30%vs.0,P=0.006,7;6.23%±2.95%vs.0,P=0.003,6;3.78%±2.18%vs.0,P=0.008,1).TLR4,nuclear factor kappa-B p65,and tumor necrosis factor-αexpression in the aorta tissue of the AS group were upregulated,whereas occludin and zonula occludens-1 expression in colon tissues was downregulated.Additionally,metabolomics identified significant differences in 21 metabolites in the feces of the AS group compared to the NC group,with further analysis linking these differences to amino acid metabolism.Conclusions:The Tibetan miniature pig model of early AS induced by high-fat intake displayed pronounced chronic inflammation.Preliminary findings suggest that the underlying mechanisms may be associated with the LPS/TLR4 pathway and intestinal metabolic disorders.展开更多
目的基于肠肾轴理论探讨补肾排毒胶囊作用于肠道屏障调节内毒素(LPS)/Toll样受体4(Toll-like receptors 4,TLR4)信号通道对慢性肾脏病(CKD)肾脏的保护作用。方法通过单侧输尿管结扎法建立慢性肾脏病模型。分为空白组(Nomel,NC)、模型组(...目的基于肠肾轴理论探讨补肾排毒胶囊作用于肠道屏障调节内毒素(LPS)/Toll样受体4(Toll-like receptors 4,TLR4)信号通道对慢性肾脏病(CKD)肾脏的保护作用。方法通过单侧输尿管结扎法建立慢性肾脏病模型。分为空白组(Nomel,NC)、模型组(Model)、补肾排毒胶囊低剂量组(Low dose group of Bushen Paidu capsule,BSPD-L)、补肾排毒胶囊中剂量组(Medium dose group of Bushen Paidu capsule,BSPD-M)、补肾排毒胶囊高剂量组(High dose group of Bushen Paidui capsule,BSPD-H)、盐酸贝那普利组(Benazepril Hydrochloride group,BH),每日给药1次,28d后取材。HE染色、Mosson染色观察肾脏组织病理改变;ELISA检测半胱氨酸蛋白酶抑制剂C(Cys C)、硫酸吲哚酚(IS)、生长转化因子-β(TGF-β)、白介素-6(IL-6)含量;生化检测血尿素氮(BUN)、血清肌酐(Scr)、血尿酸(UA)含量;Western blot检测肾、肠组织TLR4蛋白表达水平;RT-PCR检测肾、肠组织TLR4mRNA表达水平;鲎试剂盒动态浊度法内毒素定量检测。结果与NC组相比,Model组大鼠肾小球结构破坏,纤维化明显,视野中基本无完整的肾小球存在,肾小管存在明显的蛋白管型,肾小管上皮细胞脱落,Cys C、IS、TGF-β、IL-6、BUN、Scr、UA含量显著升高(P<0.01),肾、肠组织TLR4蛋白表达水平明显升高(P<0.05),肾、肠组织TLR4mRNA表达水平显著升高(P<0.01),内毒素水平显著增高(P<0.01);与Model组相比,BH、BSPD-L、BSPD-M和BSPD-H能够改善CKD的病理变化,降低Cys C、IS、TGF-β、IL-6、BUN、Scr、UA含量,下调肾、肠组织TLR4蛋白和基因表达水平,改善内毒素水平,其中BSPD-H组和BH组病理恢复最好(P<0.01)。结论补肾排毒胶囊能够作用于肠道屏障抑制LPS/TLR4信号通道,下调炎症因子水平,降低尿毒症毒素含量,改善CKD大鼠肾组织病理形态。展开更多
基金funded by the National Natural Science Foundation of China(31970514)Basic Public Welfare Projects in Zhejiang Province(LGD20C040004)the Research Project of Zhejiang Chinese Medical University(2021JKZKTS031B).
文摘Objective:This study aimed to elucidate the effect of the lipopolysaccharides/toll-like receptor 4(LPS/TLR4)pathway on early atherosclerosis(AS)development and its associated changes in fecal metabolites,thereby providing an experimental foundation for strategies to prevent and treat early AS.Methods:Twelve Tibetan miniature pigs aged 4-5 months were divided into normal control(NC)group and AS group(6 pigs in each).The group assignment was primarily based on body weight;Secondary criteria,including glucose,lipid profiles,and inflammatory indices,were considered to ensure balanced baseline characteristics between the 2 groups(all P>0.05).AS group received a high-fat diet for 16 weeks to establish an AS model,while the NC group received a normal diet.Subsequently,serum levels of lipids and various inflammation and oxidative stress markers were measured.Pathological changes in the aorta and colon tissue,LPS/TLR4 pathway-associated protein expressions in the aorta,as well as occludin and zonula occludens-1 in the colon were also assessed.Proton nuclear magnetic resonance spectra technology was employed for the metabolomic analysis of fecal extracts.Results:The lipid metabolism was disrupted in AS group,with significantly higher total cholesterol,high-density lipoprotein cholesterol,and low-density lipoprotein cholesterol levels((12.24±5.24)mmol/L vs.(1.86±0.27)mmol/L,P=0.004,6;(2.39±0.50)mmol/L vs.(0.83±0.07)mmol/L,P=0.000,5;(6.94±2.87)mmol/L vs.(0.77±0.18)mmol/L,P=0.003,3),as compared to that in NC group.Serum factors,including LPS,tumor necrosis factor-α,and malondialdehyde levels of AS group were significantly higher than that of NC group((1,230.00±192.70)EU/L vs.(695.70±213.70)EU/L),P=0.001,1;(424.20±176.90)ng/L vs.(51.20±26.61)ng/L,P=0.023,5;(3.60±0.77)nmol/mL vs.(2.62±0.21)nmol/mL,P=0.025,4).Pathological evaluations revealed prominent lipid deposition area in the aortic arch,thoracic aorta,and abdominal aorta of the AS group compared with that of the NC group(4.17%±2.30%vs.0,P=0.006,7;6.23%±2.95%vs.0,P=0.003,6;3.78%±2.18%vs.0,P=0.008,1).TLR4,nuclear factor kappa-B p65,and tumor necrosis factor-αexpression in the aorta tissue of the AS group were upregulated,whereas occludin and zonula occludens-1 expression in colon tissues was downregulated.Additionally,metabolomics identified significant differences in 21 metabolites in the feces of the AS group compared to the NC group,with further analysis linking these differences to amino acid metabolism.Conclusions:The Tibetan miniature pig model of early AS induced by high-fat intake displayed pronounced chronic inflammation.Preliminary findings suggest that the underlying mechanisms may be associated with the LPS/TLR4 pathway and intestinal metabolic disorders.
文摘目的基于肠肾轴理论探讨补肾排毒胶囊作用于肠道屏障调节内毒素(LPS)/Toll样受体4(Toll-like receptors 4,TLR4)信号通道对慢性肾脏病(CKD)肾脏的保护作用。方法通过单侧输尿管结扎法建立慢性肾脏病模型。分为空白组(Nomel,NC)、模型组(Model)、补肾排毒胶囊低剂量组(Low dose group of Bushen Paidu capsule,BSPD-L)、补肾排毒胶囊中剂量组(Medium dose group of Bushen Paidu capsule,BSPD-M)、补肾排毒胶囊高剂量组(High dose group of Bushen Paidui capsule,BSPD-H)、盐酸贝那普利组(Benazepril Hydrochloride group,BH),每日给药1次,28d后取材。HE染色、Mosson染色观察肾脏组织病理改变;ELISA检测半胱氨酸蛋白酶抑制剂C(Cys C)、硫酸吲哚酚(IS)、生长转化因子-β(TGF-β)、白介素-6(IL-6)含量;生化检测血尿素氮(BUN)、血清肌酐(Scr)、血尿酸(UA)含量;Western blot检测肾、肠组织TLR4蛋白表达水平;RT-PCR检测肾、肠组织TLR4mRNA表达水平;鲎试剂盒动态浊度法内毒素定量检测。结果与NC组相比,Model组大鼠肾小球结构破坏,纤维化明显,视野中基本无完整的肾小球存在,肾小管存在明显的蛋白管型,肾小管上皮细胞脱落,Cys C、IS、TGF-β、IL-6、BUN、Scr、UA含量显著升高(P<0.01),肾、肠组织TLR4蛋白表达水平明显升高(P<0.05),肾、肠组织TLR4mRNA表达水平显著升高(P<0.01),内毒素水平显著增高(P<0.01);与Model组相比,BH、BSPD-L、BSPD-M和BSPD-H能够改善CKD的病理变化,降低Cys C、IS、TGF-β、IL-6、BUN、Scr、UA含量,下调肾、肠组织TLR4蛋白和基因表达水平,改善内毒素水平,其中BSPD-H组和BH组病理恢复最好(P<0.01)。结论补肾排毒胶囊能够作用于肠道屏障抑制LPS/TLR4信号通道,下调炎症因子水平,降低尿毒症毒素含量,改善CKD大鼠肾组织病理形态。
