醋甘遂与炙甘草配伍对癌性腹水模型大鼠细胞色素氧化酶基因和蛋白表达的影响  

Effect of Compatibility of Vinegar-processed Euphorbia Kansui and Radix Glycyrrhizae Preparata on Cytochrome Oxidase Gene and Protein Expression in Cancerous Ascites Model Rats

作　　者：刘艳蕊 郝蕾 王婷婷 王芮 杨伟丽 王茜 LIU Yanrui;HAO Lei;WANG Tingting;WANG Rui;YANG Weili;WANG Xi(School of Pharmacy,Hebei University of Traditional Chinese Medicine,Shijiazhuang 050091 Hebei,China)

机构地区：[1]河北中医学院药学院,河北石家庄050091

出　　处：《中药新药与临床药理》2021年第11期1640-1647,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81503268);河北省中医药管理局项目(2021105);中央财政公共卫生专项“2019年全国第四次中药资源普查项目——河北省”(Z135080000022)。

摘　　要：目的通过检测癌性腹水模型大鼠肝功能指标、血清生物标志物含量及肝细胞色素P450酶(cytochrome P450)CYP2D6-1、CYP3A1、CYP2E1的基因、蛋白表达,探讨醋甘遂与炙甘草配伍对癌性腹水大鼠CYP450酶的调节作用,从而揭示二者配伍的作用实质。方法将雄性Wistar大鼠按体质量随机分为正常组、模型组、炙甘草组(免煎颗粒剂)、醋甘遂组(研末)、炙甘草-醋甘遂组(草遂组)、甘遂半夏汤组。除正常组外,其余各组腹腔注射Walker-256肿瘤细胞株复制癌性腹水模型,连续给药14 d。实验结束后取材,用生化法检测天门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、碱性磷酸酶(AKP)活性以及总胆红素(TBIL)、直接胆红素(DBIL)含量,用酶联免疫吸附法(ELISA)检测谷氨酸脱氢酶(GLDH)、苹果酸脱氢酶(MDH1)、嘌呤核苷磷酸化酶(PNP)、对氧磷酶(PON1)含量,HE染色观察肝组织病理形态学变化,免疫组织化学法检测细胞色素氧化酶CYP3A1、CYP2E1蛋白表达,采用RT-qPCR技术检测CYP2D6-1、CYP3A1、CYP2E1的基因表达。结果与正常组比较,模型组AST、ALT、TBIL、DBIL、AKP、GLDH、MDH1、PNP水平明显升高(P<0.05),PON1水平明显下降(P<0.05);与模型组比较,炙甘草组、草遂组、甘遂半夏汤组PON1含量明显升高(P<0.05)。RT-qPCR及免疫组织化学结果显示CYP2D6-1、CYP3A1、CYP2E1基因及蛋白表达与正常组比较明显升高(P<0.05);与模型组比较,各给药组均能下调CYP2D6-1、CYP3A1、CYP2E1的基因或蛋白表达(P<0.05)。HE染色结果显示,与模型组比较,炙甘草组、草遂组及甘遂半夏汤组肝索紊乱减少,肝血窦扩张减少。结论本实验提示醋甘遂与炙甘草只有在特定条件下,运用中医的辨证论治思维配伍应用才能起到减毒的作用。醋甘遂与炙甘草配伍可能通过抑制细胞色素氧化酶的基因、蛋白表达,提高PON1含量,降低氧化应激,起到对肝脏的保护作用。Objective By detecting liver function indexes,serum biomarker content,gene and protein expression of cytochrome P450(CYP2 D6-1,CYP3 A1,CYP2 E1)in cancerous ascites model rats,we discussed the regulatory effect of compatibility of vinegar-processed Euphorbia kansui and Radix Glycyrrhizae preparata on CYP450 enzyme in cancerous ascites rats,and revealed the essence of compatibility between these two herbs.Methods Male Wistar rats were randomly divided into normal group,model group,Radix Glycyrrhizae preparata group,vinegarprocessed Euphorbia kansui group,compatibilities of vinegar-processed Euphorbia kansui and Radix Glycyrrhizae preparata group(Caosui group) and Gansui Banxia decoction group according to body mass.Except the normal group,the other groups were given Walker-256 tumor cell line by intraperitoneal injection to form ascites.On the second day,the corresponding medicine liquid was given by gavage for 14 days.The activities of AST,ALT,AKP,TBIL and DBIL were detected by biochemical method,and GLDH,MDH1,PNP,PON1 were detected by enzyme-linked immunosorbent assay(ELISA).HE staining was used to observe the pathomorphological changes of liver tissue,immunohistochemical method was used to detect the protein expressions of cytochrome oxidase CYP3 A1 and CYP2 E1,real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the gene expressions of CYP2 D6-1,CYP3 A1 and CYP2 E1 in liver tissue.Results Compared with the normal group,the levels of AST,ALT,TBIL,DBIL,AKP,GLDH,MDH1 and PNP in the model group were significantly increased(P<0.05),while the levels of PON1 were significantly decreased(P<0.05).Compared with the model group,the levels of PON1 in Radix Glycyrrhizae preparata group,Caosui group and Gansui Banxia decoction group were significantly increased(P<0.05).The results of RT-qPCR and immunohistochemistry showed that the expressions of CYP2 D6-1,CYP3 A1 and CYP2 E1 genes and proteins were significantly higher than those of normal group(P<0.05).Compared with model group,the expre

关 键 词：十八反 醋甘遂 炙甘草 配伍关系 细胞色素氧化酶 大鼠 

分 类 号：R285.5[医药卫生—中药学]