基于NF-κB/NLRP3信号通路探讨栀子豉汤对酒精性肝损伤的保护作用

Protective effect of Zhi-zi-chi decoction on alcoholic liver injury based on NF-κB/NLRP3 signaling pathway

中文关键词:  栀子豉汤  酒精性肝损伤  NF-κB/NLRP3信号通路  氧化应激反应  炎症反应

英文关键词:Zhi-zi-chi decoction  alcoholic liver disease  NF-κB/NLRP3 signaling pathway  oxidative stress response  inflammatory response

中文摘要:

      目的 探讨栀子豉汤对酒精性肝损伤(alcoholic liver disease, ALD)小鼠的保护作用及对NF-κB/NLRP3信号通路的影响。方法 将60只雄性小鼠随机均分为6组: 空白组、模型组、甘草酸二铵阳性对照组(60 mg/kg)和栀子豉汤高、中、低(910.0、455.0、227.5 mg/kg) 3个剂量组。除空白组外, 其余5组均采用50%乙醇(10 mL/kg)灌胃诱导小鼠ALD模型。持续14 d。检测小鼠血清中肝功能指标水平、血清中炎症因子含量; 同时检测血清中抗氧化因子含量。HE染色观察肝脏的组织病理学改变。用Western blot检测小鼠肝组织中核苷酸寡聚化结构域样受体家族3 (nucleotide oligomeric domain-like receptor family 3, NLRP3)、半胱天冬酶-1 (caspase-1)、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a card, ASC)、核转录因子-κB(P65) [nuclear factor kappa-B p65, NF-κB(P65)]、NF-κB(PP65)、白介素-1β (interleukin-1, IL-1β)的相对表达量。结果 与模型组小鼠相比, 栀子豉汤各剂量能够显著降低小鼠肝功能指标水平(P<0.05、P<0.01、P<0.001、P<0.0001); 炎症因子含量均显著降低(P<0.01、P<0.001、P<0.0001); 血清中抗氧化能力显著提高(P<0.05、P<0.01、P<0.001、P<0.0001); 病理切片结果表明, 栀子豉汤能改善小鼠肝组织病理变化。肝组织中NLRP3、caspase-1、ASC、NF-κB(P65)、NF-κB(PP65)、IL-1β的相对表达量均显著降低(P<0.0001或P<0.001或P<0.01)。结论 栀子豉汤能改善ALD模型组小鼠肝损伤, 抑制氧化应激和炎症反应, 其作用机制可能与调控肝组织中NF-κB/NLRP3信号通路的活性有关。

英文摘要:

      Objective To investigate the protective effects of Zhi-zi-chi decoction on alcoholic liver disease (ALD) mice and its effects on NF-κB/NLRP3 signaling pathway. Methods Sixty male mice were randomly and equally divided into 6 groups: blank group, model group, diammonium glycyrrhizate positive control group (60 mg/kg) and 3 dose groups of Zhi-zi-chi decoction high, medium and low (910.0, 455.0 and 227.5 mg/kg). Except for the blank group, all 5 groups were induced with 50% ethanol (10 mL/kg) to induce the mouse ALD model by gavage for 14 d. The levels of liver function indexes, inflammatory factors and antioxidant factors in serum of mice were measured. The histopathological changes in the liver were observed by HE staining. Western blot was used to detect nucleotide oligomeric domain-like receptor family 3 (NLRP3), caspase-1, apoptosis-associated speck-like protein containing a card (ASC), nuclear factor kappa-B(P65) [NF-κB(P65)], NF-κB(PP65), interleukin-1 (IL-1β) were expressed in relative amounts. Results Compared with mice in the model group, each dose of Zhi-zi-chi decoction could significantly reduce the level of liver function indexes in mice (P<0.05, P<0.01, P<0.001, P<0.0001); inflammatory factor content was significantly reduced (P<0.01, P<0.001, P<0.0001); antioxidant capacity in serum was significantly improved (P<0.05, P<0.01, P<0.001, P<0.0001). Pathological section results showed that Zhi-zi-chi decoction could improve liver tissue pathological changes in mice. The relative expressions of NLRP3, caspase-1, ASC, NF-κB(P65), NF-κB(PP65), and IL-1β in liver tissues were significantly reduced (P<0.0001 or P<0.001 or P<0.01). Conclusion Zhi-zi-chi decoction can improve liver injury in ALD model group mice, inhibit oxidative stress and inflammatory response. Its mechanism of action may be related to regulating the activity of NF-κB/NLRP3 signaling pathway in liver tissue.