沙棘多糖提取物对扑热息痛诱导的小鼠肝损伤的保护作用及其机制研究

发布时间：2018-04-26 07:54

本文选题：沙棘多糖 + 扑热息痛　；参考：《内蒙古农业大学》2017年硕士论文

【摘要】：药物性肝损伤(Drug-induced liver injury,DILI)是由药物本身及其代谢产物所引发的肝脏损伤,是引起急性肝衰竭的主要原因。沙棘果被认为在藏医和蒙医中具有很高的药用价值,可治疗多种疾病,但对于沙棘果多糖提取物(Seabucthorn berry po1ysaccharide,SP)治疗 DILI 的研究甚少。本实验通过模拟扑热息痛(Acetaminophen,APAP)诱导的小鼠药物性肝损伤过程,探究沙棘多糖对药物性肝损伤的保护作用及其机制,为开发绿色保肝新药提供科学依据。C57BL/6雄性小鼠随机分为六组,即空白对照组(Ctrl),APAP模型组(APAP),N-乙酰半胱氨酸组(NAC),沙棘多糖低剂量组(APAP/SP100),沙棘多糖高剂量组(APAP/SP200),沙棘多糖对照组(SP200),沙棘多糖低、高剂量组及对照组分别以100、200、200mg.kg-1 SP连续灌胃30天后,腹腔注射扑热息痛(350mg.kg-1)构建DILI模型,NAC(150mg.kg-1)于建模前1h腹腔注射。16h后处死小鼠,收集血清及肝脏样本,分别检测血清中ALT和AST水平及肝脏中SOD、MDA、iNOS、GSH和GSH-PX等含量;HE染色观察肝脏组织病理损伤情况;Real-time PCR检测肝组织中细胞因子的表达水平;Western blot检测肝组织中 TLR4、p-JNK、及 p62、Keap-1、Bcl-2、Bax、Nrf-2 和 SOD-2的蛋白表达情况;分别采用Real-time PCR及Western blot检测Nrf-2及其下游靶基因HO-1、GCLC和NQO-1的mRNA及蛋白水平变化。结果表明,沙棘多糖显著降低APAP小鼠血清中ALT和AST水平。沙棘多糖提高了肝脏中SOD、GSH和GSH-PX的水平,降低了 MDA、NO和iNOS的水平。HE染色结果也表明沙棘多糖能有效缓解APAP诱导的肝毒性。沙棘多糖抑制了 TLR4、p-JNK的表达并上调了 Bcl-2/Bax。沙棘多糖抑制了 APAP小鼠中炎性细胞因子TNF-α和IL-6的表达并促进IL-10的表达;进一步研究结果表明,沙棘多糖下调Keap-1的表达、促进Nrf-2核转位及其下游靶基因HO-1和SOD-2的活化。本研究表明,沙棘多糖能够保护APAP诱导的肝损伤。该保护作用与其抗炎和抗氧化功能有关。沙棘多糖通过调控TLR4-p-JNK以及Nrf-2/HO-l-SOD-2信号通路发挥保肝作用。本研究为开发沙棘多糖类绿色药物奠定基础。
[Abstract]:Drug-induced liver injury (DILI) is a liver injury caused by the drug itself and its metabolites. It is the main cause of acute liver failure. The seabuckthorn fruit is considered to be of high medicinal value in Tibetan and Mongolian medicine, and can be used to treat a variety of diseases, but the polysaccharide extract of seabuckthorn fruit (Seabucthorn berry po1ysac) Charide, SP) little study on the treatment of DILI. This experiment was conducted to explore the protective effect and mechanism of seabuckthorn polysaccharides on drug induced liver injury induced by Acetaminophen (APAP) induced liver injury in mice and to provide a scientific basis for the development of new green liver protection drugs for the development of.C57BL/6 male mice randomly divided into six groups, that is, blank control Group (Ctrl), APAP model group (APAP), N- acetyl cysteine group (NAC), Hippophae rhamnoides polysaccharide low dose group (APAP/SP100), seabuckthorn polysaccharide high dose group (APAP/SP200), seabuckthorn polysaccharide control group (SP200), seabuckthorn polysaccharide low, high dose group and control group with 100200200mg.kg-1 SP 30 days after continuous perfusion of 100200200mg.kg-1 SP, intraperitoneal injection of paracetamol (350mg.kg-1) to construct DILI The model, NAC (150mg.kg-1) was killed in 1h after.16h injection, and the serum and liver samples were collected. The levels of serum ALT and AST and the content of SOD, MDA, iNOS, GSH and GSH-PX in the liver were detected, and the pathological damage of liver tissue was observed by HE staining. The protein expressions of TLR4, p-JNK, and p62, Keap-1, Bcl-2, Bax, Nrf-2 and SOD-2 were measured in the liver tissue, and Real-time PCR and Western blot and its downstream target genes were used respectively. The results showed that the Seabuckthorn polysaccharides decreased significantly in the serum of mice and the increase of seabuckthorn polysaccharide The level of SOD, GSH and GSH-PX in the liver decreased the level of MDA, NO and iNOS. The results also showed that the Seabuckthorn polysaccharide could effectively alleviate the liver toxicity induced by APAP. The Seabuckthorn polysaccharide inhibited the TLR4, the p-JNK expression and the inhibition of the expression of inflammatory cytokines and the expression of inflammatory cytokines in APAP mice. The further study shows that Seabuckthorn polysaccharides downregulate the expression of Keap-1, promote the Nrf-2 nuclear transposition and the activation of the downstream target gene HO-1 and SOD-2. This study shows that the Seabuckthorn polysaccharide can protect the liver injury induced by APAP. The protective effect is related to its anti-inflammatory and antioxidant functions. Seabuckthorn polysaccharides can be controlled by TLR4-p-JNK and Nrf-2/HO- L-SOD-2 signaling pathway plays a role in liver protection. This study lays the foundation for developing Seabuckthorn polysaccharide green drugs.

【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

【参考文献】