Wang Si-Wei, Lan Tian, Zheng Fang, Lei Mei-Kang, Zhang Feng
Quzhou People's Hospital Quzhou 324000,China.
Quzhou Customs Technical Center Quzhou 324000,China.
Zhongguo Zhong Yao Za Zhi. 2021 Mar;46(6):1474-1479. doi: 10.19540/j.cnki.cjcmm.20201201.401.
To study the effect and mechanism of extract of Quzhou Aurantii Fructus(QAF) on liver inflammation in CCl_4-induced liver fibrosis mice. Totally 60 C57 BL/6 male mice were randomly divided into control group(distilled water, oral), model group(distilled water, oral), colchicines group(Col, colchicines 2 mg·kg(-1)·d(-1), oral), low-dose QAF group(QAF-L, QAF 100 mg·kg(-1)·d(-1), oral) and high-dose QAF group(QAF-H, QAF 300 mg·kg(-1)·d(-1), oral) by random number table method. The model group and each administration group were injected with carbon tetrachloride(CCl_4) 1 mL·kg~(-1)(CCl_4-olive oil 1∶4), twice a week, totally 6 weeks. After the last administration, the mice were sacrificed, and serum and liver tissue were collected. Serum ALT and AST levels were measured in each group to observe the liver function of mice. The pathological changes and inflammatory cell infiltration in liver were observed by HE staining and F4/80 immunohistochemical staining. The mRNA expressions of TNF-α, IL-18 and IL-1β were detected by RT-PCR. The protein expressions of IκBα, p-IKKα/β, p-p65, NLRP3, caspase-1 and cleaved caspase-1 were analyzed by Western blot. The results showed that QAF significantly reduced serum ALT and AST levels, and alleviated the degree of liver damage.The results of immunohistochemistry showed that QAF significantly reduced liver inflammatory cell infiltration in liver fibrosis mice. The results of RT-PCR and Western blot showed that QAF significantly inhibited mRNA expressions of TNF-α, IL-18 and IL-1β in liver of fibrosis mice. QAF also suppressed the degradation of IκBα protein and reduced p-IKKα/β, p-p65, NLRP3 and cleaved caspase-1 protein expressions. In conclusion, QAF improves CCl_4-induced liver fibrosis in mice. The mechanism may be related to the inhibition of NF-κB/NLRP3 inflammasome-mediated inflammation signaling pathway.
研究衢州枳壳提取物（QAF）对四氯化碳（CCl₄）诱导的肝纤维化小鼠肝脏炎症的影响及其机制。将60只C57 BL/6雄性小鼠通过随机数字表法随机分为对照组（蒸馏水，灌胃）、模型组（蒸馏水，灌胃）、秋水仙碱组（Col，秋水仙碱2 mg·kg⁻¹·d⁻¹，灌胃）、低剂量QAF组（QAF-L，QAF 100 mg·kg⁻¹·d⁻¹，灌胃）和高剂量QAF组（QAF-H，QAF 300 mg·kg⁻¹·d⁻¹，灌胃）。模型组和各给药组每周两次腹腔注射1 mL·kg⁻¹四氯化碳（CCl₄-橄榄油1∶4），共6周。末次给药后，处死小鼠，收集血清和肝组织。检测各组血清谷丙转氨酶（ALT）和谷草转氨酶（AST）水平，观察小鼠肝功能。通过苏木精-伊红（HE）染色和F4/80免疫组化染色观察肝脏病理变化和炎性细胞浸润情况。采用逆转录-聚合酶链反应（RT-PCR）检测肿瘤坏死因子-α（TNF-α）、白细胞介素-18（IL-18）和白细胞介素-1β（IL-1β）的mRNA表达。采用蛋白质免疫印迹法分析IκBα、磷酸化IκB激酶α/β（p-IKKα/β）、磷酸化核转录因子-κB p65亚基（p-p65）、NLRP3炎性小体、半胱天冬酶-1（caspase-1）和裂解的半胱天冬酶-1（cleaved caspase-1）的蛋白表达。结果显示，QAF可显著降低血清ALT和AST水平，减轻肝脏损伤程度。免疫组化结果显示，QAF可显著减轻肝纤维化小鼠肝脏炎性细胞浸润。RT-PCR和蛋白质免疫印迹结果显示，QAF可显著抑制纤维化小鼠肝脏中TNF-α、IL-18和IL-1β的mRNA表达。QAF还可抑制IκBα蛋白降解，降低p-IKKα/β、p-p65、NLRP3和裂解的caspase-1蛋白表达。综上所述，QAF可改善CCl₄诱导的小鼠肝纤维化。其机制可能与抑制核转录因子-κB（NF-κB）/NLRP3炎性小体介导的炎症信号通路有关。
