余甘子冻干粉拮抗镉亚急性暴露致小鼠肝损伤的作用及机制  

Role and Mechanism of Freeze-Dried Powder of Phyllanthus emblica L.in Antagonizing Liver Injury Induced by Subacute Cadmium Exposure in Mice

作　　者：赵波 谷仕艳 梅宗钦 李汶泓 马顺蓉 申开颜 何作顺 ZHAO Bo;GU Shiyan;MEI Zongqin;LI Wenhong;MA Shunrong;SHEN Kaiyan;HE Zuoshun(School of Public Health,Dali University,Dali 671000;Institute of Preventive Medicine,Dali University,Dali 671000)

机构地区：[1]大理大学公共卫生学院,云南大理671000 [2]大理大学预防医学研究所,云南大理671000

出　　处：《食品科学》2025年第6期164-171,共8页Food Science

基　　金：云南省科技厅科技计划项目(202204BP090017)。

摘　　要：目的:探讨余甘子冻干粉(freeze-dried powder of Phyllanthus emblica L.,PEFP)拮抗氯化镉(CdCl_(2))致小鼠亚急性肝损伤的作用,并通过转录组学解析可能的调控机制。方法:将雄性KM小鼠随机分为对照组、PEFP组(300 mg/kg)、CdCl_(2)组(3 mg/kg)和PEFP+CdCl_(2)组(300 mg/kg PEFP+3 mg/kg CdCl_(2)),6只/组,分别灌胃PEFP(每天一次)和腹腔注射CdCl_(2)(隔天一次),持续28 d。解剖后检测肝组织镉含量、血清谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)和碱性磷酸酶(alkaline phosphatase,ALP)以及肝脏组织中的超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,Gpx)的水平,并采用苏木精-伊红(hematoxylin-eosin staining,HE)染色观察肝脏组织细胞形态,TUNEL法测定肝细胞凋亡情况,RNA测序技术确定差异表达基因,并采用蛋白免疫印迹法(Western blotting,WB)对部分基因编码的蛋白水平进行检测。结果:与对照组相比,镉单独暴露组小鼠体质量增长率((13.97±2.06)%)显著下降(P<0.05),肝脏镉蓄积量((3.49±0.09)μg/kg)增加(P<0.05),血清AST、ALT和ALP水平((707.55±60.19)、(614.58±24.40)、(186.85±12.51)U/L)升高(P<0.05),肝细胞凋亡率((32.89±2.53)%)增加(P<0.05),镉联合PEFP处理后可明显逆转镉引起的上述变化(P<0.05)。RNA测序结果显示,PEFP干预处理后有69个基因出现显著差异表达(14个上调、55个下调),这些差异表达的基因编码的蛋白与炎症、氧化损伤和凋亡有关。选择其中4个(AnnexinA2、S100A11、Mcp1、Galectin3)进行WB检测,结果与RNA测序结果一致,提示这些蛋白参与了PEFP拮抗镉的过程。结论:PEFP对镉致小鼠亚急性肝损伤有拮抗作用,可能是通过抑制炎症反应、氧化损伤和凋亡等改善肝脏组织损伤。Objective:To investigate the effect of freeze-dried powder of Phyllanthus emblica L.(PEFP)on cadmium chloride(CdCl_(2))-induced subacute liver injury in mice and to analyze the underlying mechanism by transcriptomics.Methods:Male KM mice were randomly divided into four groups with 6 animals per group:control,PEFP(300 mg/kg),CdCl_(2)(3 mg/kg)and PEFP+CdCl_(2)(300 mg/kg PEFP+3 mg/kg CdCl_(2)).PEFP was gavaged once a day to mice,and CdCl_(2) was intraperitoneally injected into mice every other day.The administration period lasted for 28 days.After euthanasia,liver tissues were collected for the determination of cadmium content,superoxide dismutase(SOD),and glutathione peroxidase(Gpx)activities,and serum aspartate aminotransferase(AST),alanine aminotransferase(ALT),alkaline phosphatase(ALP)activities were measured.Additionally,hepatic cell morphology was observed by hematoxylin-eosin(HE)staining,the apoptosis rate was determined by the terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick end labeling(TUNEL)assay,differentially expressed genes were determined by RNA sequencing,and protein expression levels encoded of the selected differentially expressed genes were detected by Western blot(WB).Results:Compared with the control group,body mass gain rate significantly decreased in the CdCl_(2) exposure group((13.97±2.06)%)(P<0.05),liver cadmium accumulation((3.49±0.09)μg/kg),and serum AST,ALT and ALP levels((707.55±60.19),(614.58±24.40),and(186.85±12.51)U/L,respectively)and hepatocyte apoptosis rate((32.89±2.53)%)increased(P<0.05).However,CdCl_(2) combined with PEFP treatment could significantly reverse the above changes caused by cadmium(P<0.05).RNA sequencing showed that 69 genes were differentially expressed after PEFP intervention(14 up-regulated and 55 down-regulated),the encoded protein of which were associated with inflammation,oxidative damage and apoptosis.We selected four proteins(AnnexinA2,S100A11,Mcp1,and galectin3)encoded of these differentially expressed genes for further verification by WB

关 键 词：氯化镉 余甘子冻干粉 肝损伤 氧化应激 凋亡 炎症 

分 类 号：R151[医药卫生—营养与食品卫生学]