摘要
目的探究罗沙司他(ROX)对2型糖尿病(T2DM)大鼠糖脂代谢、肝功能、炎性反应、肝脏脂肪蓄积及Toll样受体4(TLR4)/核转录因子-κB(NF-κB)通路的影响。方法于2024年8月—2025年1月在长江航运总医院动物实验室进行实验。T2DM大鼠采用饲喂高脂高糖饲料和腹腔注射链脲佐菌素造模,将T2DM大鼠60只按随机数字表法分成:模型组、ROX低剂量组(12.5 mg/kg)、ROX中剂量组(25 mg/kg)、ROX高剂量组(50 mg/kg)、盐酸二甲双胍组(200 mg/kg),每组12只。另取12只正常大鼠给予普通饲料喂养及腹腔一次性注射生理盐水作为对照组。各组给予相应干预8周。全自动生化仪检测大鼠血糖、血脂、肝功能指标,并测定肝指数;酶联免疫吸附法检测血清炎性因子水平;HE染色和油红O染色观察肝组织病理学及脂肪蓄积;蛋白印迹法检测肝组织TLR4/NF-κB通路蛋白表达。结果与对照组比较,模型组大鼠肝组织结构紊乱,肝细胞形态改变,出现脂质空泡和大片橘红色,脂肪蓄积程度严重,且血清空腹血糖(FPG)、空腹胰岛素(FINS)、胰岛素抵抗指数(HOMA-IR)、总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、肝指数、C-反应蛋白(CRP)、肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)、肝组织TLR4、磷酸化NF-κB p65(p-NF-κB p65)/NF-κB p65蛋白表达升高,高密度脂蛋白胆固醇(HDL-C)降低(均P<0.01);与模型组比较,ROX低、中、高剂量组及盐酸二甲双胍组大鼠肝组织病变明显好转,肝细胞中橘红色面积减小,脂肪蓄积程度减轻,且血清FPG、FINS、HOMA-IR、TC、TG、LDL-C、ALT、AST、肝指数、CRP、TNF-α、IL-6、肝组织TLR4、p-NF-κB p65/NF-κB p65蛋白表达降低,HDL-C升高(均P<0.01),ROX低、中、高剂量组血清FPG、FINS、HOMA-IR、TC、TG、LDL-C、ALT、AST、肝指数、CRP、TNF-α、IL-6、肝组织TLR4、p-NF-κB p65/NF-κB p65蛋白表达降低程度及HDL-C升高程度呈现剂量依赖性(均P<0.01);ROX高剂量组和盐酸二甲双胍组大鼠肝组织结构形态相近,趋于正常,肝细胞中橘红色面积相近,脂肪蓄积程度减轻最为明显,糖脂代谢指标、肝功能指标及肝指数、炎性因子、肝组织TLR4/NF-κB通路蛋白表达差异无统计学意义(P>0.05)。结论ROX能够改善T2DM大鼠糖脂代谢、肝功能及炎性反应,减轻肝组织病理损伤和脂肪蓄积,并抑制TLR4/NF-κB通路。
Objective To explore roxadustat's effects on diabetes-associated metabolic/liver dysfunction.Methods The experiments were conducted at the Animal Laboratory of the Changjiang Shipping General Hospital from August 2024 to January 2025.T2DM rats were induced by feeding high fat and high sugar feed and intraperitoneal injection of streptozotocin.60 T2DM rats were randomly divided into:model group,ROX low dose group(12.5 mg/kg),ROX medium dose group(25 mg/kg),ROX high dose group(50 mg/kg),metformin hydrochloride group(200 mg/kg),12 animals in each group;another 12 normal rats were given normal feed for 8 weeks and normal saline was injected into the abdominal cavity as healthy control group;each group was given corresponding intervention for 8 weeks.Automatic biochemical analyzer was used to detect the blood glucose,blood lipids and liver function indexes of rats,and the liver index was determined.Serum inflammatory factors were detected by enzyme-linked immunosorbent assay.Liver tissue histopathology and fat accumulation were observed by hematoxylin-eosin staining and oil red O staining.The expression of TLR4/NF-κB pathway protein in liver tissue was detected by western blot.Results Compared with healthy control group,the liver tissue structure of rats in model group was disordered,the morphology of liver cells was changed,lipid vacuoles and large orange red areas appeared,and the degree of fat accumulation was serious;serum fasting blood glucose(FPG),fasting insulin(FINS),insulin resistance index(HOMA-IR),total cholesterol(TC),triacylglycerol(TG),low density lipoprotein-cholesterol(LDL-C),alanine aminotransferase(ALT),aspartate aminotransferase(AST),liver index,C-reactive protein(CRP),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),the expressions of TLR4,phosphorylated-NF-κB p65(p-NF-κB p65)/NF-κB p65 protein in liver tissue were increased,and high density lipoprotein-cholesterol(HDL-C)was decreased(t/P=12.983/<0.001,10.640/<0.001,19.194/<0.001,16.195/<0.001,17.406/<0.001,17.660/<0.001,14.805/<0.001,13.273/<0.001,9.039/<0.001,10.590/<0.001,16.086/<0.001,20.255/<0.001,22.789/<0.001,14.976/<0.001,9.824/<0.001).Compared with model group,the rats of ROX low,medium,and high dose groups and metformin hydrochloride group showed significant improvement in liver tissue lesions,reduced orange red area in liver cells,decreased degree of fat accumulation;serum FPG,FINS,HOMA-IR,TC,TG,LDL-C,ALT,AST,liver index,CRP,TNF-α,IL-6,TLR4,p-NF-κB p65/NF-κB p65 protein expression in liver tissue were decreased,while HDL-C was increased(F/P=37.001/<0.001,34.685/<0.001,99.532/<0.001,101.881/<0.001,120.663/<0.001,105.021/<0.001,61.833/<0.001,48.872/<0.001,37.652/<0.001,36.541/<0.001,69.113/<0.001,86.194/<0.001,66.174/<0.001,63.913/<0.001,27.611/<0.001).The degree of decrease in serum FPG,FINS,HOMA-IR,TC,TG,LDL-C,ALT,AST,liver index,CRP,TNF-α,IL-6,liver tissue TLR4,p-NF-κB p65/NF-κB p65 protein expression in ROX low,medium,high dose groups and the degree increase in HDL-C were dose-dependent(F/P=19.952/<0.001,18.082/<0.001,48.212/<0.001,63.192/<0.001,88.165/<0.001,66.031/<0.001,31.735/<0.001,27.221/<0.001,22.784/<0.001,22.072/<0.001,40.682/<0.001,48.306/<0.001,39.842/<0.001,44.825/<0.001,14.002/<0.001).The liver tissue structure and morphology of rats in ROX high dose group and metformin hydrochloride group were similar and tended to be normal,the orange area of liver cells was similar,the degree of fat accumulation was most significantly reduced,and there was no statistical significance in the glucose and lipid metabolism indexes,liver function indexes and liver index,inflammatory factors,and TLR4/NF-κB pathway protein expression in liver tissue(P>0.05).Conclusion Roxadustat ameliorates diabetic liver injury by modulating glucose metabolism and TLR4/NF-κB signaling.
作者
石秀祯
何艳
徐申
郭爱莉
付锴
谭金枚
Shi Xiuzhen;He Yan;Xu Shen;Guo Aili;Fu Kai;Tan Jinmei(Department of Nephrology,Changjiang Shipping General Hospital,Hubei,Wuhan 430014,China;不详)
出处
《疑难病杂志》
2025年第9期1115-1121,1131,共8页
Chinese Journal of Difficult and Complicated Cases
基金
湖北省卫生健康委科研立项项目(WJ2023M024)
长航局科技项目(202210005)。
