藏药波棱瓜子总木脂素对刀豆球蛋白(ConA)致免疫性肝损伤小鼠保护作用及其机制探讨

目的:波棱瓜子是常见的用于治疗肝病的藏药,疗效确切.前期研究表明,总木脂素是波棱瓜子抗肝损伤的有效部位,但疗效尚不明确.本研究拟考察藏药波棱瓜子总木脂素对ConA致小鼠免疫性肝损伤的保护作用,并进一步探讨其对抗肝损伤的作用机制.方法:采用尾静脉注射一次性ConA(20mg/kg)诱发小鼠免疫性肝损伤,造模8h后取血样和肝脏,考察不同剂量的波棱瓜子总木脂素对各组小鼠脏器系数、血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、骨过氧化物酶(MPO)、一氧化氮(NO)、一氧化氮合酶(NOS)及肝组织匀浆中超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)等生化指标活力或水平的影响;以HE染色对肝组织进行组织病理学检查;免疫组织化学法检测波棱瓜子总木脂素对各组小鼠肝脏中肿瘤坏死因子-α(TNF-α)、干扰素-γ(IFN-γ)、白细胞介素-4(IL-4)、白细胞介素-10(IL-10)、核转录因子-κB(NF-κB)表达的影响.结果:波棱瓜子总木脂素可以降低ConA所致免疫性肝损伤小鼠肝脏系数及血清ALT、AST、ALP活力,显示出较好的保肝作用.同时波棱瓜子总木脂素也可降低肝损伤小鼠血清NO水平和MPO、NOS活力,并减少肝组织匀浆中MDA的含量,增强SOD和GSH-Px活性,表现出良好的抗氧化作用;免疫组化结果显示,波棱瓜子总木脂素还能抑制诸如TNF-α、IFN-γ、IL-4、NF-κB等促炎症因子的表达,促进抗炎症因子IL-10的表达,从而抑制肝脏炎症反应而发挥其保肝作用.病理观察结果也表明,波棱瓜子总木脂素能减轻ConA对肝组织的损伤.结论:波棱瓜子总木脂素对ConA致小鼠免疫性肝损伤都具有一定的保护作用,其保肝作用可能与其抗炎、抗氧化能力有关.

The protective effect and its mechanism of total Lignans from Tibetan medicinal Herpetospermum Seeds on Concanavalin A-induced liver injury in mice

Objective： To investigate the protection effect and mechanism of the total Lignans from Tibetan Medicinal Herpetospermum Seeds on immunologic liver injury induced by ConA. Methods： Immunologic liver injury was induced by a single intravenous injection of ConA （20mg/kg）. Then animals were sacrificed at the 8h post-injection, blood samples and liver were collected. Subsequently, organ index of liver, and biochemical parameters, such as ALT, AST, ALP, NO, NOS, MPO in serum and SOD, MDA, GSH-Px in liver tissue were evaluated. The pathological changes of the liver tissue were measured by HE staining. Finally, the expressions of TNF-ct, IFN-？, IL-4, IL-10, NF-κB were tested with immunohistochemical method. Results： The total Lignans could reduce serum activities of ALT, AST, ALP, NOS, MPO and serum levels of NO. Meanwhile, lower serum levels of MDA and elevated activities of SOD and GSH-Px were observed in total Lignans treated liver injury mice. Furthermore, total Lignans significantly reduced the expressions of TNF-α, IFN-γ, IL-4 and NF-κB, and increased the expressions of IL-10. Pathological results also showed that the total Lignans could attenuate ConA-induced liver damage. Conclusion： The total Lignans from Tibetan Medicinal Herpetospermum Seeds have a certain Hepatoprotective effect on ConA-induced immunological liver injury in mice, which may be related its anti-oxidant and anti-inflammatory effects.