黑种草子提取物对小鼠酒精性肝损伤的修复作用及机制研究

doi:10.16431/j.cnki.1671-7236.2022.12.040

摘要/Abstract

摘要： 【目的】验证黑种草子提取物对小鼠酒精性肝损伤的修复作用并探究其机制，为其临床开发研究提供理论依据。【方法】将56只3周龄昆明小鼠随机均分为7组：空白对照组，模型组，黑种草子提取物高、中、低剂量组，黑种草子粉剂组及水飞蓟宾阳性对照组，每组8只。除空白对照组外，其余各组均以10 mL/kg 60%酒精灌胃，每天1次，连续15 d。第16天，黑种草子提取物高、中、低剂量组分别给予8、4、2 mL/kg黑种草子提取物，每天1次，连续10 d后处死小鼠。计算各组小鼠肝脏指数；检测血清中谷丙转氨酶（alanine aminotransferase，ALT）、谷草转氨酶（aspartate aminotransferase，AST）、过氧化氢酶（catalase，CAT）、超氧化物歧化酶（superoxide dismutase，SOD）和谷胱甘肽过氧化物酶（glutathione peroxidase，GSH-Px）活力；制备肝脏石蜡切片观察其病理变化；Western blotting检测NOD样受体热蛋白结构域相关蛋白3（NOD-like receptor thermal protein domain associated protein 3，NLRP3）、含半胱氨酸的天冬氨酸蛋白水解酶1（cysteinyl aspartate specific proteinase 1，Caspase-1）和白细胞介素1β（interleukin-1β，IL-1β）的蛋白表达水平。【结果】与空白对照组相比，模型组小鼠肝脏指数显著上升（P<0.05），肝细胞出现明显颗粒变性，水泡变性，核溶解，界限区分不清，肝索消失；血清ALT、AST活力显著上升（P<0.05），SOD、CAT、GSH-Px活力显著下降（P<0.05），说明肝损伤模型建立成功，且肝脏组织中NLRP3、Caspase-1和IL-1β蛋白表达水平显著升高（P<0.05）。与模型组相比，黑种草子提取物各剂量组血清ALT、AST活力均呈下降趋势，SOD、CAT、GSH-Px活力均呈上升趋势，其中高剂量组上述各指标均呈显著差异（P<0.05），肝脏组织结构明显改善，NLRP3、Caspase-1和IL-1β蛋白表达水平均显著下降（P<0.05）。【结论】高剂量（8 mL/kg）黑种草子提取物可显著提高肝损伤小鼠的抗氧化水平，且可以通过抑制NLRP3炎性小体的表达而减少IL-1β的合成，从而改善肝脏功能和修复受损的肝脏组织结构。

关键词: 黑种草子提取物; 小鼠; 酒精性肝损伤; NLRP3炎性小体; 白细胞介素1β (IL-1β)

Abstract: 【Objective】 This study was aimed to verify the repairing effect of Nigella sativa seed extract regarding to alcoholic liver injury in mice, and explore its mechanism, so as to provide theoretical basis for its clinical development and research.【Method】 Fifty-six 3-week-old Kunming mice were randomly divided into 7 groups:Blank control group, model group, high, medium and low dose Nigella sativa seed extract groups, Nigella sativa seed powder group and silybin positive control group, with 8 mice in each group.Except for blank control group, the mice in other groups were given 10 mL/kg 60% alcohol by gavage, once a day for 15 consecutive days.On the 16th day, the mice in high, medium and low dose Nigella sativa seed extract groups were given 8, 4 and 2 mL/kg Nigella sativa seed extract, respectively, and the mice were sacrificed for 10 consecutive days.The liver index of mice in each group was calculated, the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in serum were detected.Paraffin sections were prepared to observe the pathological changes of liver, the protein expression of NOD-like receptor thermal protein domain associated protein 3 (NLRP3), cysteinyl aspartate specific proteinase 1 (Caspase-1) and interleukin-1β (IL-1β) were detected by Western blotting.【Result】 Compared with blank control group, the liver index of mice in model group was significantly increased (P<0.05), and the liver cells showed obvious granular degeneration, vesicular degeneration, nucleus lysis, unclear differentiation boundaries and disappearance of hepatic cords.The activities of ALT and AST in serum were significantly increased (P<0.05), the activity of SOD, CAT and GSH-Px were significantly decreased (P<0.05), indicating the liver damage model successfully established, and the expression of NLRP3, Caspase-1 and IL-1β proteins in liver were significantly increased (P<0.05).Compared with model group, the activity of ALT and AST in serum showed a downward trend in three dose Nigella sativa seed extract groups, and the activity of SOD, CAT and GSH-Px showed an upward trend, among which the above indicators in high-dose group were significantly different (P<0.05), the liver tissue structure was significantly improved, and the expression of NLRP3, Caspase-1 and IL-1β proteins were decreased significantly (P<0.05).【Conclusion】 High doses of Nigella sativa seed extract (8 mL/kg) could significantly increase the antioxidant levels in liver injury mice, and reduce the synthesis of IL-1β by inhibiting the expression of NLRP3 inflammatory body, thereby improving liver function and repairing damaged liver tissue structure.

Key words: Nigella sativa seed extract; mice; alcoholic liver injury; NLRP3 inflammatory body; interleukin-1β (IL-1β)

中图分类号:

S852.3

赵伟捷, 孙耀贵, 孙盼盼, 孙娜, 杨惠珍, 李宏全. 黑种草子提取物对小鼠酒精性肝损伤的修复作用及机制研究[J]. 中国畜牧兽医, 2022, 49(12): 4928-4935.

ZHAO Weijie, SUN Yaogui, SUN Panpan, SUN Na, YANG Huizhen, LI Hongquan. Study on Repair Effect and Mechanism of Nigella sativa Seed Extract on Alcoholic Liver Injury in Mice[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(12): 4928-4935.

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