Xie Jun, Liu Jie, Chen Tu-Ming, Lan Qing, Zhang Qing-Yu, Liu Bin, Dai Dong, Zhang Wei-Dong, Hu Li-Ping, Zhu Run-Zhi
Jun Xie, Wei-Dong Zhang, Li-Ping Hu, Run-Zhi Zhu, Clinical Research Center, Xuyi People's Hospital, Xuyi 211700, Jiangsu Province, China.
World J Gastroenterol. 2015 May 14;21(18):5473-81. doi: 10.3748/wjg.v21.i18.5473.
To assess the effects of dihydromyricetin (DHM) as a hepatoprotective candidate in reducing hepatic injury and accelerating hepatocyte proliferation after carbon tetrachloride (CCl4) treatment.
C57 BL/6 mice were used in this study. Mice were orally administered with DHM (150 mg/kg) for 4 d after CCl4 treatment. Serum and liver tissue samples were collected on days 1, 2, 3, 5 and 7 after CCl4 treatment. The anti-inflammatory effect of DHM was assessed directly by hepatic histology detection and indirectly by serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), albumin, and superoxide dismutase (SOD). Inflammatory cytokines, such as interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α), were detected using ELISA kits. Proliferating cell nuclear antigen (PCNA) staining was used to evaluate the role of DHM in promoting hepatocyte proliferation. Hepatocyte apoptosis was measured by TUNEL assay. Furthermore, apoptosis proteins Caspases-3, 6, 8, and 9 were detected by Western blot. SP600125 were used to confirm whether DHM regulated liver regeneration through JNK/TNF-α pathways.
DHM showed a strong anti-inflammatory effect on CCl4-induced liver injury in mice. DHM could significantly decrease serum ALT, AST, IL-1β, IL-6 and TNF-α and increase serum albumin, SOD and liver SOD compared to the control group after CCl4 treatment (P < 0.05). PCNA results indicated that DHM could significantly increase the number of PCNA positive cells compared to the control (348.9 ± 56.0 vs 107.1 ± 31.4, P < 0.01). TUNEL assay showed that DHM dramatically reduced the number of apoptotic cells after CCl4 treatment compared to the control (365.4 ± 99.4 vs 90.5 ± 13.8, P < 0.01). Caspase activity detection showed that DHM could reduce the activities of Caspases- 8, 3, 6 and 9 compared to the control (P < 0.05). The results of Western blot showed that DHM increased the expression of JNK and decreased TNF-α expression. However, DHM could not affect TNF-α expression after SP600125 treatment. Furthermore, DHM could significantly improve the survival rate of acute liver failure (ALF) mice (73.3% vs 20.0%, P < 0.0001), and SP600125 could inhibit the effect of DHM.
These findings demonstrate that DHM alleviates CCl4-induced liver injury, suggesting that DHM is a promising candidate for reversing liver injury and ALF.
评估二氢杨梅素（DHM）作为一种肝保护候选药物在减轻四氯化碳（CCl4）处理后肝损伤及加速肝细胞增殖方面的作用。
本研究使用C57 BL/6小鼠。CCl4处理后，小鼠口服给予DHM（150 mg/kg），持续4天。在CCl4处理后的第1、2、3、5和7天收集血清和肝组织样本。通过肝脏组织学检测直接评估DHM的抗炎作用，并通过血清天冬氨酸转氨酶（AST）、丙氨酸转氨酶（ALT）、白蛋白和超氧化物歧化酶（SOD）水平间接评估。使用酶联免疫吸附测定试剂盒检测白细胞介素（IL）-1β、IL-6和肿瘤坏死因子-α（TNF-α）等炎性细胞因子。使用增殖细胞核抗原（PCNA）染色评估DHM在促进肝细胞增殖中的作用。通过TUNEL测定法检测肝细胞凋亡。此外，通过蛋白质免疫印迹法检测凋亡蛋白半胱天冬酶-3、6、8和9。使用SP600125来确认DHM是否通过JNK/TNF-α途径调节肝脏再生。
DHM对CCl4诱导的小鼠肝损伤显示出强大的抗炎作用。与CCl4处理后的对照组相比，DHM可显著降低血清ALT、AST、IL-1β、IL-6和TNF-α水平，并增加血清白蛋白、SOD以及肝脏SOD水平（P < 0.05）。PCNA结果表明，与对照组相比，DHM可显著增加PCNA阳性细胞数量（348.9 ± 56.0对107.1 ± 31.4，P < 0.01）。TUNEL测定显示，与对照组相比，DHM在CCl4处理后显著减少了凋亡细胞数量（365.4 ± 99.4对90.5 ± 13.8, P < 0.01）。半胱天冬酶活性检测表明，与对照组相比，DHM可降低半胱天冬酶-8、3、6和9的活性（P < 0.05）。蛋白质免疫印迹结果显示，DHM增加了JNK的表达并降低了TNF-α的表达。然而，SP600125处理后，DHM不影响TNF-α的表达。此外，DHM可显著提高急性肝衰竭（ALF）小鼠的存活率（73.3%对20.0%，P < 0.0001），且SP600125可抑制DHM的作用。
这些研究结果表明，DHM可减轻CCl4诱导的肝损伤，提示DHM是逆转肝损伤和ALF的有前景的候选药物。
