应用黄药子煎剂灌胃建立小鼠肝损伤模型的初步研究

发布时间：2018-08-04 16:18

【摘要】：【目的】初步探索应用肝毒性中药黄药子煎剂灌胃建立小鼠肝损伤模型，并从脂质过氧化损伤方面分析其机制。【方法】（1）昆明种清洁级小鼠40只，体重18-22g，雌雄各半。随机分为4组：空白组、黄药子煎剂高剂量组、中剂量组、低剂量组，每组10只。黄药子煎剂各组分别以240g·kg~(-1)、120g·kg~(-1)、60g·kg~(-1)，按0.2mL·10g~(-1)体重黄药子煎剂灌胃；空白组以等体积蒸馏水灌胃。每日1次，连续灌胃6d后处死，分别检测血清丙氨酸氨基转移酶（alanine aminotransferase，ALT）、天门冬氨酸氨基转移酶（aspertateaminotransferase，AST），计算肝指数，观察肝组织普通病理形态学变化，以探索黄药子煎剂致明显肝损伤的相对合适剂量。（2）昆明种清洁级小鼠30只，体重18-22g，雌雄各半。随机分为3组：空白组、黄药子煎剂给药6d组、12d组，每组10只。黄药子煎剂组以探索出的相对合适剂量黄药子煎剂，按0.2mL·10g~(-1)体重灌胃；空白组给予等体积蒸馏水灌胃。每日1次，，给药组连续灌胃6d、12d后分别处死10只，分别检测血清ALT、AST、总胆红素（total bilirubin，TBil），计算肝指数，观察肝组织普通病理形态学变化，以确定致明显肝损伤的相对合适时间。并检测小鼠肝组织丙二醛（malondialdehyde，MDA）、谷胱甘肽过氧化物酶（glutathione peroxidase，GSH-Px），初步探讨黄药子致肝损伤的机理。【结果】（1）不同剂量黄药子煎剂灌胃对小鼠血清ALT、AST、肝指数的影响：与空白组比较，各剂量组黄药子煎剂对小鼠血清ALT、AST及肝指数均有明显影响（P0.05）。高、中、低剂量三组之间两两比较，ALT的差异明显，黄药子剂量越大，ALT越高，与用药剂量呈正相关（P0.05）；低与高、中剂量组间AST的差异有显著意义（P0.05）；三组之间肝指数比较无显著统计学意义（P0.05）。（2）不同剂量黄药子煎剂灌胃对小鼠肝组织形态学的影响：低剂量组小鼠未出现明显肝损伤；中、高剂量组小鼠肝损伤明显，尤以高剂量组为甚。经肝细胞变性半定量分析，低剂量黄药子煎剂灌胃对小鼠肝细胞无明显影响（P0.005），高、中剂量组与空白组相比有显著统计学意义（P0.005）。（3）高剂量黄药子煎剂灌胃不同时间对小鼠血清ALT、AST、TBil及肝指数的影响：6d、12d组小鼠血清ALT、AST、TBil及肝指数明显升高，与空白组比较有显著的统计学意义（P0.05）。但两组之间无显著统计学差异（P0.05）。（4）高剂量黄药子煎剂灌胃不同时间对肝组织形态学的影响：6d、12d组小鼠均出现明显肝组织损伤。经肝细胞变性半定量分析，高剂量黄药子煎剂灌胃6d、12d与空白组比较均有显著统计学意义（P0.005），但两组之间比较无显著统计学意义（P0.005）。（5）高剂量黄药子煎剂灌胃6d对小鼠肝组织MDA、GSH-Px的影响：与空白组比较，高剂量黄药子煎剂灌胃6d对小鼠肝组织MDA和GSH-Px水平影响不大，无显著统计学意义（P0.05）。【结论】（1）应用高剂量（240g·kg~(-1)）黄药子煎剂灌胃6d可造成小鼠明显肝损伤。（2）自由基脂质过氧化可能没有参与黄药子煎剂诱导的肝损伤，但仍需在以后的实验中进一步确定。
[Abstract]:[Objective] To establish a model of liver injury in mice by intragastric administration of Huangyaozi decoction, a hepatotoxic Chinese herb, and analyze its mechanism from the aspect of lipid peroxidation injury.
[method]
(1) 40 clean grade mice in Kunming, weighing 18-22g, and male and female, were divided into 4 groups randomly: blank group, high dose group of xanthate decoction, middle dose group, low dose group, 10 rats in each group. Each group of xanthate decoctions was treated with 240g kg~ (-1), 120g. Kg~ (-1), 60g. 1 times a day and 1 times a day after continuous perfusion of the stomach, the serum alanine aminotransferase (alanine aminotransferase, ALT), aspartate aminotransferase (aspertateaminotransferase, AST), the liver index were calculated and the normal pathological changes of liver tissue were observed in order to explore the relative appropriate liver damage caused by the decoction of xanthate. Dose.
(2) 30 clean grade mice in Kunming, weighing 18-22g, and male and female, were randomly divided into 3 groups: blank group, 6D group of xanthate decoction, group 12D, 10 in each group. The relative suitable dosage of xanthate decoction, 0.2mL. 10g~ (-1) weight was given to the stomach. The blank group was given equal volume distilled water for gastric perfusion. 1 times a day, the group was connected to the drug group. After continuous perfusion of 6D and 12D, 10 rats were killed respectively. The serum ALT, AST, total bilirubin (total bilirubin, TBil) were detected respectively. The liver index was calculated and the normal pathological changes of liver tissue were observed to determine the appropriate time to cause obvious liver injury. And the mice liver tissue two aldehyde (malondialdehyde, MDA), glutathione peroxidase (glutathione) was detected. Peroxidase (GSH-Px), to explore the mechanism of liver injury induced by Xanthii.
[results]
(1) the effect of different doses of xanthate Decoction on the serum ALT, AST and liver index of mice: compared with the blank group, the decoction of xanthate had significant influence on the serum ALT, AST and liver index of mice (P0.05). The difference of ALT between the high, middle and low dosage groups was 22, the higher the dose of the xanthate, the higher the ALT, and the dosage of the drug. There was positive correlation (P 0.05); there was significant difference in AST between low and high dose groups (P 0.05); there was no significant difference in liver index among the three groups (P 0.05).
(2) the effect of different doses of xanthate Decoction on the histomorphology of liver in mice: there was no obvious liver injury in the low dose group; in the high dose group, the liver injury was obvious, especially in the high dose group. The low dose xanthate decoction had no obvious effect on the mice liver cells (P0.005), high dose and medium dose. There was a significant difference between the group and the blank group (P0.005).
(3) the effect of high dose xanthate Decoction on the serum ALT, AST, TBil and liver index of mice at different time: 6D, the serum ALT, AST, TBil and liver index in the group 12D mice increased significantly, and there was significant statistical significance (P0.05) compared with the blank group (P0.05), but there was no significant difference between the two groups (P0.05).
(4) the effect of high dose xanthate Decoction on the histomorphology of liver in different time: 6D, group 12D mice had obvious liver tissue damage. The semi quantitative analysis of liver cell degeneration, high dose of xanthate Decoction 6D, 12D and blank group had significant statistical significance (P0.005), but there was no significant statistical significance between the two groups (P0.00 5).
(5) the effect of high dose of xanthate Decoction on MDA and GSH-Px in liver tissue of mice: compared with the blank group, the effect of high dose of xanthate Decoction on the level of MDA and GSH-Px in liver tissues of mice was not significant (P0.05), and the effect of high dose xanthate Decoction on the level of MDA and GSH-Px in liver tissues of mice was not significant (P0.05).
[Conclusion] (1) (1) the application of high dose (240g. Kg~ (-1)) xanthate Decoction to 6D can cause obvious liver damage in mice. (2) free radical lipid peroxidation may not be involved in liver injury induced by xanthate decoction, but it still needs to be further determined in the future experiment.
【学位授予单位】：河南中医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R-332;R259

【参考文献】