黑果枸杞汁对大鼠酒精性肝损伤的保护作用*

doi:10.12047/j.cjap.6242.2022.035

中国应用生理学杂志 ›› 2022, Vol. 38 ›› Issue (3): 241-246.doi: 10.12047/j.cjap.6242.2022.035

黑果枸杞汁对大鼠酒精性肝损伤的保护作用^*

胡戈¹, 曹建民², 周海涛^3,4△, 张静^3,4, 田一鸣³, 宋滢洋³, 蒋若雨³

1.常州大学体育学院, 常州 213164;
2.北京体育大学运动人体科学学院, 北京 100084;
3.北京联合大学, 北京 100101;
4.北京联合大学生物活性物质与功能食品北京市重点实验室, 北京 100191

收稿日期:2021-12-13 修回日期:2022-04-02 出版日期:2022-05-28 发布日期:2022-09-05
通讯作者: ^△Tel: 13611383040; E-mail: zsettle@sina.com
基金资助:
^*北京市朝阳区协同创新项目(CYXC1817);北京市高等学校高水平人才交叉培养“实培计划”项目;北京联合大学科研项目(ZK70202005);北京联合大学“启明星”大学生科技创新创业项目(20211006)

Protective effects of Lycium ruthenicum Murr. juice on alcoholic liver injury in rats

HU Ge¹, CAO Jian-min², ZHOU Hai-tao^3,4△, ZHANG Jing^3,4, TIAN Yi-ming³, SONG Ying-yang³, JIANG Ruo-yu³

1. Physical Education College, Changzhou University, Changzhou 213164;
2. Sport Science College of Beijing Sport University, Beijing 100084;
3. Beijing Union University, Beijing 100101;
4. Key Laboratory of Bioactive Substances and Functional Foods, Beijing Union University, Beijing 100191, China

Received:2021-12-13 Revised:2022-04-02 Online:2022-05-28 Published:2022-09-05

摘要/Abstract

摘要： 目的: 研究黑果枸杞汁对大鼠酒精性肝损伤的保护作用,探讨其中Toll样受体4(TLR4)/p38 丝裂原活化蛋白激酶(p38 MAPK)信号通路的调节机制。方法: 60只雄性SD大鼠随机分为空白对照组(C)、模型组(M)、低剂量黑果枸杞汁组(LLM)、中剂量黑果枸杞汁组(MLM)、高剂量黑果枸杞汁组(HLM),每组12只。M、LLM、MLM和HLM组每天以20 ml/kg(8 g/(kg·d))剂量分2次灌胃400 g/L酒精,C组于相同时间点灌胃等体积蒸馏水。每日首次酒精灌胃前4 h,黑果枸杞汁各剂量组分别以2.4、4.8、9.6 ml/(kg·d)进行灌胃,其他组于相同时间点灌胃等体积蒸馏水。实验周期4周,末次实验结束后24 h处死大鼠,取血液和肝脏,计算肝脏指数,HE染色观察肝脏组织形态,比色法检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)活性,免疫组化法检测肝脏TLR4、p38 MAPK及磷酸化p38 MAPK(p-p38 MAPK)蛋白质表达,酶联免疫吸附试验检测肝脏肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-10(IL-10)、白细胞介素-18(IL-18)水平。结果: 与C组比较,M组成功建立酒精性肝损伤模型。与M组比较,黑果枸杞汁各剂量组的相关指标出现改善,其中HLM组肝脏组织形态改善最为显著,其肝脏系数、血清ALT和AST、肝脏TLR4蛋白质表达、p-p38 MAPK/p38 MAPK比值、TNF-α、IL-1β和IL-18水平均显著降低(P＜0.05或P＜0.01),肝脏IL-10水平显著升高(P＜0.01)。黑果枸杞汁各剂量组组间比较,肝脏系数、血清AST、肝脏TLR4蛋白质表达、p-p38 MAPK/p38 MAPK比值、TNF-α和IL-18水平,HLM组较LLM组均显著降低(P＜0.05或P＜0.01);肝脏IL-10水平,HLM组较LLM和MLM组均显著升高(P＜0.05或P＜0.01);其余主要指标组间比较均无统计学差异(P＞0.05)。结论: 黑果枸杞汁可以通过调控TLR4/p38 MAPK信号通路,改善炎症应激,缓解大鼠酒精性肝损伤,高剂量组效果优于其他剂量组。

关键词: 黑果枸杞汁, 酒精性肝损伤, Toll样受体4/p38 丝裂原活化蛋白激酶信号通路, 炎症, 大鼠

Abstract: Objective: To study the protective effects of Lycium ruthenicum Murr. juice on alcoholic liver injury in rats and explore the regulatory mechanism of toll-like receptors 4 (TLR4)/p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway in this process. Methods: Sixty male SD rats were randomly divided into control group (C), model group (M), low-dose Lycium ruthenicum Murr. juice group (LLM), medium-dose Lycium ruthenicum Murr. juice group (MLM) and high-dose Lycium ruthenicum Murr. juice group (HLM), 12 rats in each group. The group M, LLM, MLM and HLM were treated with 20 ml/kg (8 g/(kg·d)) ethanol (400 g/L) intragastrically and the gavage was divided into two sessions, group C was treated with an equal volume of distilled water at the same time point. Four hours before the first alcohol gavage session, rats in each dose group of Lycium ruthenicum Murr. juice were administered with 2.4, 4.8, 9.6 ml/(kg·d) Lycium ruthenicum Murr. juice respectively, and the other groups were given equal volume of distilled water at the corresponding time points. Four weeks later, the rats were sacrificed 24 hours after the end of the last experiment, blood and liver were collected. The liver index was calculated. The morphology of the liver was observed by HE staining. The expressions of hepatic TLR4, p38 MAPK and phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK) were detected by immunohistochemistry. The activities of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected by colorimetry. The levels of hepatic tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-10 (IL-10) and interleukin-18 (IL-18) were detected by enzyme linked immunosorbent assay. Results: Compared with group C, the alcoholic liver injury model was established successfully in Group M. Compared with group M, related indicators in each dose group of Lycium ruthenicum Murr. juice were improved, the improvement of hepatic morphology in group HLM was the most significant, the liver index, the levels of serum ALT, AST and hepatic TLR4, p38 MAPK/p-p38 MAPK ratio, TNF-α, IL-1β, IL-18 were decreased (P＜ 0.05 or P＜0.01), while the level of hepatic IL-10 was increased (P＜0.01). Comparison among the dose groups of Lycium ruthenicum Murr. juice, the levels of liver index, serum AST and hepatic TLR4, p38 MAPK/p-p38 MAPK ratio, TNF-α, IL-18 in HLM were lower than those in LLM (P＜0.05 or P＜0.01); the level of hepatic IL-10 in HLM was higher than that in LLM and MLM (P＜0.05 or P＜0.01); the other indicators in each dose group had no statistical difference (P＞0.05). Conclusion: Lycium ruthenicum Murr. juice can improve the inflammatory stress by regulating TLR4/p38 MAPK signaling pathway, relieve alcoholic liver injury in rats, and the effect of high-dose group is better than the others.

Key words: Lycium ruthenicum Murr. juice, alcoholic liver injury, toll-like receptors 4/p38 mitogen-activated protein kinase signaling pathway, inflammation, rat

中图分类号:

G804.7

胡戈, 曹建民, 周海涛, 张静, 田一鸣, 宋滢洋, 蒋若雨. 黑果枸杞汁对大鼠酒精性肝损伤的保护作用^*[J]. 中国应用生理学杂志, 2022, 38(3): 241-246.

HU Ge, CAO Jian-min, ZHOU Hai-tao, ZHANG Jing, TIAN Yi-ming, SONG Ying-yang, JIANG Ruo-yu. Protective effects of Lycium ruthenicum Murr. juice on alcoholic liver injury in rats[J]. CJAP, 2022, 38(3): 241-246.

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黑果枸杞汁对大鼠酒精性肝损伤的保护作用^*

Protective effects of Lycium ruthenicum Murr. juice on alcoholic liver injury in rats

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