摘要
目的 探索慈姑多糖(Sagittaria Sagittifolia polysaccharide, SSP)是否通过介导内质网应激的蛋白激酶R样内质网激酶(protein kinase RNA-like endoplasmic reticulum kinase, PERK)通路对6种重金属联合所致小鼠肝损伤发挥保护作用。方法 首先根据北京市售大米6种重金属膳食摄入比例,构建6种重金属混合染毒液所致小鼠肝损伤体内模型。40只雄性昆明小鼠分为5组:对照组,损伤模型组,谷胱甘肽阳性对照组(900 mg/kg)和SSP低、高剂量组(200、800 mg/kg),每组8只。灌胃干预30 d,每5 d称量小鼠体质量,计算体质量变化量。苏木素-伊红(HE)染色观察小鼠肝脏损伤情况;全自动生化分析仪检测小鼠血清白蛋白(albumin, ALB)含量和碱性磷酸酶(alkaline phosphatase, ALP)活性;试剂盒检测血清中丙二醛(malondialdehyde, MDA)、谷胱甘肽(glutathione, GSH)含量和超氧化物歧化酶(superoxide dismutase, SOD)活性;Western blot法检测肝组织中核因子κB(nuclear factor kappa B,NF-κB)、免疫球蛋白重链结合蛋白(immunoglobulin heavy chain binding protein, BiP)、PERK、C/EBP同源蛋白(C/EBP homologous protein, CHOP)的蛋白表达情况;免疫组织化学法检测BiP、PERK蛋白表达量;DAPI染色观察肝细胞凋亡情况。结果 HE染色结果显示,与模型组相比,SSP治疗组肝脏中炎症细胞浸润及细胞坏死显著减轻,其中高剂量组损伤改善更为明显。与模型组相比,SSP治疗组中ALP活性、MDA含量显著降低(P<0.01),ALB、GSH含量和SOD活性升高(P<0.01),NF-κB p65、BiP、PERK和CHOP蛋白表达显著降低(P<0.01),肝细胞凋亡显著改善。结论 SSP可有效改善6种重金属联用诱导的小鼠肝损伤,可能与调控内质网应激的PERK通路、抑制氧化损伤和炎症反应所致细胞凋亡相关。
Objective To explore whether Sagittaria Sagittifolia polysaccharide(SSP)plays a protective role in mice injured by six heavy metals mixture by regulating the protein kinase RNA-like endoplasmic reticulum kinase(PERK)signaling pathway of endoplasmic reticulum stress(ERS).s Methods Firstly,according to the dietary intake of six heavy metals in rice sold in Beijing,a heavy metal mixture was prepared for inducing liver injury in Kunming mice.Forty male Kunming mice were randomly divided into 5 groups:control group,model group,glutathione positive control group(900 mg/kg),SSP low-dose and high-dose groups(200,800 mg/kg),with 8 mice in each group.The mice were weighed every 5 days and the intervention by gavage lasted for 30 days.HE staining was used to observe the pathological changes of liver injury in mice.Albumin(ALB)and alkaline phosphatase(ALP)were measured by automatic biochemical analyzer.The levels of malondialdehyde(MDA),glutathione(GSH)and activity of superoxide dismutase(SOD)were detected by using assay kits.The protein expressions of nuclear factor kappa B(NF-κB),immunoglobulin heavy chain binding protein(BiP),PERK and C/EBP homologous protein(CHOP)were detected by Western blot and immunohistochemical analysis.DAPI staining was used to observe the hepatocyte apoptosis.s Results HE staining showed that compared with those of the model group,the inflammatory cell infiltration and cell necrosis in liver were significantly im-proved in the SSP treatment groups,and the effect was more obvious in the SSP high-dose group.Compared with those of the model group,the ALP activity and MDA content in the SSP treatment groups decreased(P<0.01),the levels of GSH and ALB and the SOD activity significantly increased(P<0.01),the protein expressions of NF-κB p65,BiP,PERK and CHOP decreased significantly(P<0.01),and the hepatocyte apoptosis improved significantly.n Conclusion SSP can effectively improve the liver in-jury induced by the mixture of six heavy metals,and its underlying mechanism may be related to the regulation
作者
周漫钰
王萌
刘红双
高鑫
刘冰清
张烯
廖艳
ZHOU Manyu;WANG Meng;LIU Hongshuang;GAO Xin;LIU Bingqing;ZHANG Xi;LIAO Yan(School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 102446,China;Functional Food Engineering Technology in Shaanxi Research Center,Xi’an 710054,Shaanxi,China)
出处
《中华中医药学刊》
CAS
北大核心
2023年第10期135-139,I0025-I0027,共8页
Chinese Archives of Traditional Chinese Medicine
基金
国家重点研发计划项目(2018YFC1706800)。
关键词
慈姑多糖
多种重金属
内质网应激
氧化应激
Sagittaria Sagittifolin polysaccharide
multiple heavy metals
endoplasmic reticulum stress
oxidative stress
