吲哚硒对急性肝损伤的保护作用及其可能机制
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摘要
目的：探讨吲哚硒(indole-selenium,IND-Se)对四氯化碳(carbon tetrachloride,CCl_4)致小鼠急性化学性肝损伤和卡介苗(Bacillus Calmette Guerin,BCG)加脂多糖(lipopolysaccharide,LPS)诱导的小鼠免疫性肝损伤模型的保护作用及可能机制，并与同等剂量的褪黑素(melatonin,MT)或硒(selenium,Se)进行药效学比较。
方法：分别建立小鼠化学性和免疫性肝损伤模型，分光光度法检测血清中丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate
aminotransferase,AST)、一氧化氮(nitric oxide,NO)水平和肝匀浆中丙二醛(malondiadehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-px)含量，放免法检测血清和腹腔巨噬细胞(peritoneal macrophages,PM(?))培养上清液中肿瘤坏死因子-α(tumor necrosis
factor-alpha,TNF-α)含量，MTT法检测PM(?)培养上清液中白细胞介素-1(interleukin-1,IL-1)的含量。HE染色法对肝脏组织作病理检查。
结果：(1)IND-Se(5、10、20mg·kg~(-1))灌胃给药能降低化学性肝损伤小鼠血清中升高的转氨酶水平，病理检查也发现其可明显减轻肝组织坏死范围及程度，减少炎细胞浸润；同时发现IND-Se可降低肝匀浆中升高的MDA水平，使降低的肝匀浆SOD、GSH-px活性升高。(2)IND-Se(5、10、20mg·kg~(-1))灌胃给药能降低免疫性肝损伤小鼠血清中升高的转氨酶水平，病理检查也发现其可明显减轻肝组织坏死范围及程度，减少炎细胞浸润；进一步研究发现IND-Se可降低肝匀浆中升高的MDA水平，使降低的肝匀浆SOD、GSH-px活性升高，同时还发现IND-Se可降低血清中升高的NO和TNF-α水平，抑制免疫性肝损伤小鼠PM Φ TNF-α和IL-1的产生。(3)IND-Se(10mg·kg~(-1))组与MT(10mg·kg~(-1))和Se(0.1mg·kg~(-1))组相比较，除了对转氨
安徽医科大学硕士学位论文
酶水平、MDA的生成和GSH一px活性的作用的差异具有显著性外，而对其余指标的
影响无显著性差异。结论:INO一se对化学性肝损伤和免疫性肝损伤小鼠均具有保打’
作用，其机制与其抗自由基、提高抗氧化物酶的活性和抑制PM中产生TNF一a和IL一I
等有关。
Objective To investigate the effects of indole-selenium on carbon tetrachloride-induced chemical liver injury and Bacillus Calmette Guerin (BCG) plus lipopolysaccharide(LPS)-induced immunological liver injury in mice as well as its possible mechanisms. At the same time the pharmacodynamics of indole-selenium was compared with melatonin or selenium of equal dose.
Methods The model of chemical liver injury and immunological liver injury in mice was prepared. The levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST) , nitric oxide(NO) in serum, malondiadehyde(MDA) content, superoxide dismutase(SOD) and glutathione peroxidase(GSH-px) activities in liver homogenate were assayed by spectrophotometry; Tumour necrosis factor-a (TNF-a ) activity was determined by ridio immunoassay method. The level of interleukin-1(lL-1) was assayed by MTT dye reduction. Meanwhile, hepatic pathological examination was observed.
Results (1) Indole-selenium (5,10,20mg kg-1) was able to significantly decrease serum transaminase (ALT, AST) levels of chemical liver injury's mice induced by carbon tetrachloride, attenuate the area and extent of necrosis and reduce the infiltration of inflammatory cells. Furthermore, indole-selenium also decreased MDA content and improved the reduced SOD and GSH-px levels in liver homogenate.(2) Indole-selenium (5,10,20mg kg-1) was able to significantly decrease serum transaminase (ALT, AST) levels of immunological liver injury's mice induced by BCG plus LPS, attenuate the area and extent of necrosis and reduce the infiltration of inflammatory cells. Furthermore,
indole-selenium also decreased MDA content and improved the reduced SOD and GSH-px levels in liver homogenate. Meanwhile, indole-selenium significantly lowered increased TNF- and NO level in serum and inhibited the production of TNF- a and IL-1 by peritoneal macrophages. (3) Comparing with MT(10 mg kg-1) and Se(0.1 mg kg-1), Indole-selenium ( 10mg kg-1) significantly further changed the serum transaminase levels, MDA production and GSH-px activity while had no difference effect on other index.
Conclusion Indole-selenium possesses protective action on chemical liver injury and immunological liver injury in mice, which was related to direct with free radical scavenging, increasing the content of SOD and GSH-px, inhibiting the production of TNF- a and IL-1 by peritoneal macrophages and so on.
方法：分别建立小鼠化学性和免疫性肝损伤模型，分光光度法检测血清中丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate
aminotransferase,AST)、一氧化氮(nitric oxide,NO)水平和肝匀浆中丙二醛(malondiadehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-px)含量，放免法检测血清和腹腔巨噬细胞(peritoneal macrophages,PM(?))培养上清液中肿瘤坏死因子-α(tumor necrosis
factor-alpha,TNF-α)含量，MTT法检测PM(?)培养上清液中白细胞介素-1(interleukin-1,IL-1)的含量。HE染色法对肝脏组织作病理检查。
结果：(1)IND-Se(5、10、20mg·kg~(-1))灌胃给药能降低化学性肝损伤小鼠血清中升高的转氨酶水平，病理检查也发现其可明显减轻肝组织坏死范围及程度，减少炎细胞浸润；同时发现IND-Se可降低肝匀浆中升高的MDA水平，使降低的肝匀浆SOD、GSH-px活性升高。(2)IND-Se(5、10、20mg·kg~(-1))灌胃给药能降低免疫性肝损伤小鼠血清中升高的转氨酶水平，病理检查也发现其可明显减轻肝组织坏死范围及程度，减少炎细胞浸润；进一步研究发现IND-Se可降低肝匀浆中升高的MDA水平，使降低的肝匀浆SOD、GSH-px活性升高，同时还发现IND-Se可降低血清中升高的NO和TNF-α水平，抑制免疫性肝损伤小鼠PM Φ TNF-α和IL-1的产生。(3)IND-Se(10mg·kg~(-1))组与MT(10mg·kg~(-1))和Se(0.1mg·kg~(-1))组相比较，除了对转氨
安徽医科大学硕士学位论文
酶水平、MDA的生成和GSH一px活性的作用的差异具有显著性外，而对其余指标的
影响无显著性差异。结论:INO一se对化学性肝损伤和免疫性肝损伤小鼠均具有保打’
作用，其机制与其抗自由基、提高抗氧化物酶的活性和抑制PM中产生TNF一a和IL一I
等有关。
Objective To investigate the effects of indole-selenium on carbon tetrachloride-induced chemical liver injury and Bacillus Calmette Guerin (BCG) plus lipopolysaccharide(LPS)-induced immunological liver injury in mice as well as its possible mechanisms. At the same time the pharmacodynamics of indole-selenium was compared with melatonin or selenium of equal dose.
Methods The model of chemical liver injury and immunological liver injury in mice was prepared. The levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST) , nitric oxide(NO) in serum, malondiadehyde(MDA) content, superoxide dismutase(SOD) and glutathione peroxidase(GSH-px) activities in liver homogenate were assayed by spectrophotometry; Tumour necrosis factor-a (TNF-a ) activity was determined by ridio immunoassay method. The level of interleukin-1(lL-1) was assayed by MTT dye reduction. Meanwhile, hepatic pathological examination was observed.
Results (1) Indole-selenium (5,10,20mg kg-1) was able to significantly decrease serum transaminase (ALT, AST) levels of chemical liver injury's mice induced by carbon tetrachloride, attenuate the area and extent of necrosis and reduce the infiltration of inflammatory cells. Furthermore, indole-selenium also decreased MDA content and improved the reduced SOD and GSH-px levels in liver homogenate.(2) Indole-selenium (5,10,20mg kg-1) was able to significantly decrease serum transaminase (ALT, AST) levels of immunological liver injury's mice induced by BCG plus LPS, attenuate the area and extent of necrosis and reduce the infiltration of inflammatory cells. Furthermore,
indole-selenium also decreased MDA content and improved the reduced SOD and GSH-px levels in liver homogenate. Meanwhile, indole-selenium significantly lowered increased TNF- and NO level in serum and inhibited the production of TNF- a and IL-1 by peritoneal macrophages. (3) Comparing with MT(10 mg kg-1) and Se(0.1 mg kg-1), Indole-selenium ( 10mg kg-1) significantly further changed the serum transaminase levels, MDA production and GSH-px activity while had no difference effect on other index.
Conclusion Indole-selenium possesses protective action on chemical liver injury and immunological liver injury in mice, which was related to direct with free radical scavenging, increasing the content of SOD and GSH-px, inhibiting the production of TNF- a and IL-1 by peritoneal macrophages and so on.
引文
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