Hu Ge, Cao Jian-Min, Zhou Hai-Tao, Zhang Jing, Tian Yi-Ming, Song Ying-Yang, Jiang Ruo-Yu
Physical Education College, Changzhou University, Changzhou 213164.
Sport Science College of Beijing Sport University, Beijing 100084.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2022 Sep;38(3):241-246. doi: 10.12047/j.cjap.6242.2022.035.
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. 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. 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 (＜ 0.05 or ＜0.01), while the level of hepatic IL-10 was increased (＜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 (＜0.05 or ＜0.01); the level of hepatic IL-10 in HLM was higher than that in LLM and MLM (＜0.05 or ＜0.01); the other indicators in each dose group had no statistical difference (＞0.05). 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.
研究黑果枸杞汁对大鼠酒精性肝损伤的保护作用，并探讨Toll样受体4（TLR4）/p38丝裂原活化蛋白激酶（p38 MAPK）信号通路在此过程中的调控机制。将60只雄性SD大鼠随机分为对照组（C）、模型组（M）、黑果枸杞汁低剂量组（LLM）、黑果枸杞汁中剂量组（MLM）和黑果枸杞汁高剂量组（HLM），每组12只。M组、LLM组、MLM组和HLM组采用20 ml/kg（8 g/(kg·d)）乙醇（400 g/L）灌胃，分两次进行，C组在相同时间点给予等体积蒸馏水。在首次酒精灌胃前4小时，黑果枸杞汁各剂量组大鼠分别给予2.4、4.8、9.6 ml/(kg·d)黑果枸杞汁，其他组在相应时间点给予等体积蒸馏水。4周后，在最后一次实验结束24小时后处死大鼠，采集血液和肝脏。计算肝脏指数。通过HE染色观察肝脏形态。采用免疫组织化学法检测肝组织中TLR4、p38 MAPK和磷酸化p38丝裂原活化蛋白激酶（p-p38 MAPK）的表达。采用比色法检测血清丙氨酸氨基转移酶（ALT）和天冬氨酸氨基转移酶（AST）活性。采用酶联免疫吸附测定法检测肝组织肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、白细胞介素-10（IL-10）和白细胞介素-18（IL-18）水平。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相关指标均有改善，HLM组肝脏形态改善最明显，肝脏指数、血清ALT、AST水平及肝组织TLR4、p38 MAPK/p-p38 MAPK比值、TNF-α、IL-1β、IL-18水平降低（＜0.05或＜0.01），而肝组织IL-10水平升高（＜0.01）。黑果枸杞汁各剂量组比较，HLM组肝脏指数、血清AST水平及肝组织TLR4、p38 MAPK/p-p38 MAPK比值、TNF-α、IL-18水平低于LLM组（＜0.05或＜0.01）；HLM组肝组织IL-10水平高于LLM组和MLM组（＜0.05或＜0.01）；各剂量组其他指标差异无统计学意义（＞0.05）。黑果枸杞汁可通过调节TLR4/p38 MAPK信号通路改善炎症应激，减轻大鼠酒精性肝损伤，高剂量组效果优于其他组。
