基于肝脏药物代谢酶CYP450表达探讨鸦胆子苦醇对裸鼠的肝毒性研究

doi:10.12092/j.issn.1009-2501.2025.08.005

摘要/Abstract

摘要： 目的：基于肝脏药物代谢酶CYP450表达探讨具有广谱抗癌效应的鸦胆子苦醇对裸鼠的肝毒性研究。方法：取56只裸鼠随机分为空白组、鸦胆子苦醇低剂量组（2mg/kg）、鸦胆子苦醇高剂量组（4mg/kg）、顺铂组（2mg/kg），每组14只，各组使用相应药物进行腹腔注射，空白组使用等量生理盐水注射，每3d注射1次，持续6周。计算各组裸鼠死亡率情况，观察裸鼠常规生长状态，记录给药前后裸鼠体质量变化情况，取材后称量记录肝脏重量，并计算肝脏系数（肝脏重量/体质量×100%），观察记录肝脏颜色、形态；苏木素-伊红（HE）染色观察肝脏组织病理变化；ELISA检测裸鼠血清中的丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、乳酸脱氢酶（LDH）、碱性磷酸酶（AKP）和白蛋白（ALB）水平；实时荧光定量聚合酶链式反应（Real-timePCR）和Westernblot分别检测裸鼠肝脏药物代谢关键酶 CYP450 中的多个亚型酶 CYP2E1、CYP3A11、 CYP2C19、CYP1A2、CYP2D6和CYP2C9的mRNA及蛋白表达水平。结果：与空白组比较，鸦胆子苦醇低剂量组裸鼠死亡率为0，生长状态良好，饮食、行动、精神状态正常，体质量变化情况、肝脏系数比一致，肝脏颜色红润，镜下可见肝小叶形态完整，结构清晰，肝细胞排列规则，无炎症细胞浸润，ALT、AST、 LDH、AKP、ALB 含量无统计学差异，CYP2E1、 CYP3A11、CYP2C19、CYP1A2、CYP2D6 和 CYP2C9 的 mRNA及蛋白表达无统计学差异（均为P>0.05）；鸦胆子苦醇高剂量组裸鼠死亡率为14.3%，生长状态略微欠佳，饮食、行动、精神状态减低，体质量增长缓慢，肝脏系数比增大，肝脏颜色为红褐色，部分肝小叶界限不清，少量肝细胞排列松散，ALT、AST、 LDH、AKP、ALB 含量显著增高，CYP2E1、CYP3A11、 CYP1A2、CYP2D6 和 CYP2C9 的mRNA水平显著降低，CYP2E1、CYP3A11、CYP1A2和CYP2D6的蛋白表达显著降低（均为P<0.05或P<0.01），但CYP2C19的 mRNA及蛋白表达、CYP2C9的蛋白表达无统计学差异（P>0.05）；顺铂组裸鼠死亡率为35.7%，生长状态较差，饮食、行动、精神状态低下，体质量增长较少，肝脏系数比显著增高，肝脏颜色为暗红色，肝窦和中央静脉充血，肝细胞排列紊乱、胞核固缩，排列松散，ALT、AST、LDH、AKP、ALB含量显著增高，CYP2E1、 CYP3A11、CYP2C19、CYP1A2、CYP2D6 和 CYP2C9 的 mRNA及蛋白表达显著降低（均为P<0.05或P< 0.01）。结论：鸦胆子苦醇的剂量与对裸鼠的肝毒性具有相关性，高剂量的鸦胆子苦醇对裸鼠具有肝毒性，可能与降低血清中ALT、AST、LDH、AKP、ALB水平，抑制肝脏药物代谢关键酶CYP450中的多个亚型酶的表达进而降低毒性物质的代谢有关。

关键词: 鸦胆子苦醇, 肝毒性, 药物代谢酶, 裸鼠, 顺铂

Abstract: AIM: Hepatotoxicity of Brucea javani ca picryl with broad-spectrum anticancer effect in nude mice based on hepatic drug metabolizing en zyme CYP450 activity. METHODS: Fifty-six nude mice were randomly divided into blank group, Bru cea javanica low-dose group (2 mg/kg), Brucea ja vanica high-dose group (4 mg / kg), and cisplatin weight after taking materials, and calculate the liv er coefficient (liver weight/weight mass×100%), ob serve and record the liver color and morphology. Hematoxylin-eosin (HE) staining was used to ob serve the pathological changes of liver tissue. De tection of alanine aminotransferase (ALT), aspar tate aminotransferase (AST), lactate dehydrogenase (LDH), alkaline phosphatase (AKP) and albumin (ALB) levels in serum of nude mice by ELISA. Real time PCR and Western blot were used to detect the mRNA and protein expression levels of CYP2E1, CYP3A11, CYP2C19, CYP1A2, CYP2D6 and CYP2C9, which were key enzymes of drug metabolism in nude mice liver. RESULTS: Compared with the blank group, the mortality rate of nude mice in the low dose Brucea javanica bitter alcohol group was 0, the growth state was good, the diet, movement, and mental state were normal, the weight change and liver coefficient ratio were consistent, the liver color was ruddy, the liver lobule morphology was complete under the microscope, the structure was clear, the liver cells were arranged regularly, and there was no inflammatory cell infiltration. There was no significant difference in the content of ALT, AST, LDH, AKP, and ALB. There was no significant difference in the mRNA and protein expression of CYP2E1, CYP3A11, CYP2C19, CYP1A2, CYP2D6, and CYP2C9 (all P>0.05). Compared with the blank group, the mortality rate of nude mice in the high dose group of Brucea javanica bitter alcohol was 14.3%, the growth state was slightly poor, the diet, movement, and mental state were reduced, the weight growth was slow, the liver coefficient ratio was increased, the liver color was reddish brown, some liver lobule boundaries were unclear, a small group (2 mg/kg), with 14 mice in each group. The blank group was injected with the same amount of normal saline every 3 days for 6 weeks.Calculate the mortality rate of nude mice in each group, ob serve the general growth state of nude mice, re cord the weight change of nude mice before and af ter administration, weigh and record the liver number of liver cells were loosely arranged, the contents of ALT, AST, LDH, AKP, and ALB were signif icantly increased, the mRNA levels of CYP2E1, CYP3A11, CYP2C19, CYP1A2, CYP2D6, and CYP2C9 were significantly reduced, and the protein expres sions of CYP2E1, CYP3A11, CYP1A2, and CYP2D6 were significantly reduced (all P < 0.05 or P < 0.01), but there was no statistical difference in the mRNA and protein expression of CYP2C19, and the pro tein expression of CYP2C9 (P>0.05).Compared with the blank group, the mortality rate of nude mice in the cisplatin group was 35.7%, the growth state was poor, the diet, action, and mental state were low, the weight gain was less, the liver coefficient ratio was significantly increased, the liver color was dark red, the liver sinusoids and central veins were congested, the hepatocytes were disordered, the nuclei were consolidated and contracted, and the arrangement was loose, the contents of ALT, AST, LDH, AKP, and ALB were significantly increased, and the mRNA and protein expressions of CYP2E1, CYP3A11, CYP2C19, CYP1A2, CYP2D6, and CYP2C9 were significantly reduced (all P < 0.05 or P < 0.01). CONCLUSION: The dose of Brucea javanica bitter alcohol is correlated with hepatotoxicity to nude mice. High doses of Brucea javanica bitter alcohol have hepatotoxicity to nude mice, which may be re lated to reducing serum levels of ALT, AST, LDH, AKP, and ALB, inhibiting the expression of multiple subtypes of enzymes in the key enzyme CYP450 of liver drug metabolism, and then reducing the me tabolism of toxic substances.

Key words: Brucea javanica bitter alcohol, hepa totoxicity, drug metabolizing enzymes, nude mice, cisplatin

中图分类号:

R965.2

邹红, 祁硕, 李丹丹, 邓芳萍, 陈双双, 符舒欣, 唐政, 唐群. 基于肝脏药物代谢酶CYP450表达探讨鸦胆子苦醇对裸鼠的肝毒性研究[J]. 中国临床药理学与治疗学, 2025, 30(8): 1049-1057.

ZOU Hong, QI Shuo, LI Dandan, DENG Fangping, CHEN Shuangshuang, FU Shuxin, TANG Zheng, TANGQun. Hepatotoxicity of Brucea javanica bitter alcohol in nude mice based on liver drug metabolizing enzyme CYP450 expression[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2025, 30(8): 1049-1057.

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