莲心总碱对脂多糖/D-氨基半乳糖诱导急性肝衰竭小鼠肝脏炎症反应的缓解作用

doi:10.7506/spkx1002-6630-201805033

食品科学 ›› 2018, Vol. 39 ›› Issue (5): 220-225.doi: 10.7506/spkx1002-6630-201805033

莲心总碱对脂多糖/D-氨基半乳糖诱导急性肝衰竭小鼠肝脏炎症反应的缓解作用

张浪，郝吉，黄旭，吴超群，冯天辉，舒广文*

中南民族大学药学院民族药学国家级实验教学示范中心，湖北武汉 430074

出版日期:2018-03-15 发布日期:2018-03-14
基金资助:
湖北省高等学校省级教学研究项目（2015195）

Total Alkaloids from the Seed Embryo of Nelumbo nucifera Ameliorate Inflammation in the Livers of Mice with Lipopolysaccharide/D-Galactosamine-Induced Acute Liver Failure

ZHANG Lang, HAO Ji, HUANG Xu, WU Chaoqun, FENG Tianhui, SHU Guangwen*

National Demonstration Center for Experimental Ethnopharmacology Education, School of Pharmaceutical Sciences, South-Central University for Nationalities, Wuhan 430074, China

Online:2018-03-15 Published:2018-03-14

摘要/Abstract

摘要： 目的：观察莲心总碱（total alkaloids from the seed embryo of Nelumbo nucifera，TAENN）对脂多糖（lipopolysaccharide，LPS）/D-氨基半乳糖（D-galactosamine，GalN）诱导急性肝衰竭小鼠肝脏炎症反应的影响。方法：将昆明小鼠40 只随机分为4 组（正常对照组、模型组和TAENN高、低剂量组），每组10 只。TAENN高、低剂量组的给药剂量分别为100、30 mg/kg，给药途径为灌胃，每日给药1 次。正常对照组和模型组灌胃等体积生理盐水。给药结束后，模型组和TAENN高、低剂量组小鼠腹腔注射LPS/GalN建立小鼠急性肝衰竭模型。用酶联免疫吸附测定法检测肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-1β（interleukin-1β，IL-1β）、 IL-6和前列腺素E2（prostaglandine E2，PGE2）水平。用Griess法检测肝组织中一氧化氮（nitric oxide，NO）的含量。用实时聚合酶链式反应（polymerase chain reaction，PCR）法检测小鼠肝组织中TNF-α、IL-1β、IL-6、诱导型一氧化氮合酶（inducible nitric oxide synthase，iNOS）、环氧合酶-2（cyclooxygenase-2，COX-2）的mRNA的表达水平。用免疫印迹法检测小鼠肝组织中iNOS、COX-2蛋白表达和核转录因子-κB（nuclear transcription factor-κB， NF-κB）p65亚基磷酸化水平。结果：与模型组相比，TAENN组小鼠血清与肝脏中炎症因子TNF-α、IL-1β和IL-6水平明显降低（P＜0.01）。肝脏中炎症介质NO和PGE2水平亦有极显著下降（P＜0.01）。经TAENN处理后，模型小鼠肝脏中TNF-α、IL-1β、IL-6、iNOS、COX-2等的mRNA表达水平明显下降（P＜0.01）。此外，iNOS和COX-2的蛋白表达水平明显降低（P＜0.01），p65蛋白本身的表达水平不变，但其536位丝氨酸的磷酸化水平亦极显著下降（P＜0.01）。结论：TAENN对LPS/GalN诱导小鼠急性肝衰竭相关的炎症反应具有明显的保护作用，其机制可能与抑制NF-κB信号通路并减少炎症因子和炎症介质相关基因的表达有关。

关键词: 莲心总碱, 脂多糖/D-氨基半乳糖, 急性肝衰竭, 炎症

Abstract: Objective: To investigate the effect of total alkaloids from the seed embryo of Nelumbo nucifera (TAENN) on inflammation in the livers of mice with acute liver failure (ALF) induced by lipopolysaccharide (LPS)/D-galactosamine (GalN). Methods: Kunming mice were randomly divided into four groups including normal control, ALF model, TAENN low-dosage (30 mg/kg) and TAENN high-dosage groups (100 mg/kg). The drug was administered by gavage once a day. After 2 weeks, the model group and two TAENN groups were intraperitoneally injected with LPS and GalN. The contents of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, and prostaglandine E2 (PGE2) were determined by enzyme linked immunosorbent assay (ELISA). NO contents in livers were examined by Griess’ method. The transcription of TNF-α, IL-1β, IL-6, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in livers were analyzed by real-time PCR. The protein expression levels of iNOS and COX-2, and the phosphorylation levels of nuclear transcription factor -κB (NF-κB) p65 subunit at Ser536 in mouse livers were detected by immunoblotting. Results: TAENN declined the levels of TNF-α, IL-1β and IL-6 (P < 0.01) in the sera and livers of ALF model mice. In the livers of ALF model mice, the contents of inflammatory mediators such as NO and PGE2 were significantly decreased (P < 0.01) in response to TAENN. Moreover, TAENN does-dependently declined the transcription of the encoding genes such as TNF-α, IL-1β, IL-6, iNOS and COX-2 (P < 0.01) in livers. Consistently, the protein expression levels of iNOS and COX-2 were greatly reduced by TAENN (P < 0.01). Although the protein expression levels of p65 remained unchanged, the phosphorylation of p65 at Ser536 was remarkably inhibited after TAENN treatment (P < 0.01). Conclusion: TAENN inhibited inflammatory responses in the livers of mice with LPS/GalN-induced ALF. This anti-inflammation effect was associated with restrained NF-κB signaling and reduced expression of inflammatory cytokines and mediators.

Key words: total alkaloids from the seed embryo of Nelumbo nucifera, lipopolysaccharide/D-galactosamine, acute liver failure, inflammation

中图分类号:

R285.5

张浪，郝吉，黄旭，吴超群，冯天辉，舒广文. 莲心总碱对脂多糖/D-氨基半乳糖诱导急性肝衰竭小鼠肝脏炎症反应的缓解作用[J]. 食品科学, 2018, 39(5): 220-225.

ZHANG Lang, HAO Ji, HUANG Xu, WU Chaoqun, FENG Tianhui, SHU Guangwen. Total Alkaloids from the Seed Embryo of Nelumbo nucifera Ameliorate Inflammation in the Livers of Mice with Lipopolysaccharide/D-Galactosamine-Induced Acute Liver Failure[J]. FOOD SCIENCE, 2018, 39(5): 220-225.

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莲心总碱对脂多糖/D-氨基半乳糖诱导急性肝衰竭小鼠肝脏炎症反应的缓解作用

Total Alkaloids from the Seed Embryo of Nelumbo nucifera Ameliorate Inflammation in the Livers of Mice with Lipopolysaccharide/D-Galactosamine-Induced Acute Liver Failure

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