火麻仁油藻油组合物与丹参山楂三七组合物联用对代谢相关脂肪性肝病模型小鼠的改善作用及机制探讨

Study on Improvement and Mechanism of Hemp Seed Oil and Algal Oil Mixture Combined with Salviae Miltiorrhizae Radix et Rhizoma-Crtaegi Fructus-Notoginseng Radix et Rhizoma for Treating Metabolic Associated Fatty Liver Disease

目的 探讨火麻仁油藻油组合物(商品名:悦通凝胶果糖,简称YT)与丹参山楂三七组合物(商品名:怡瑞胶囊,简称YR)在小鼠代谢相关脂肪性肝病(MAFLD)中的改善作用和分子机制。方法 采用高脂饮食(High Fat Diet,HFD)诱导C57BL/6J小鼠高脂模型6周,随后干预组分别采用YT、YR和高、低剂量YT+YR灌胃给药10周。16周后,检测各组小鼠血清脂质沉积、肝功能、炎症因子和氧化应激水平,分析肝脏组织病变程度,并采用RT-PCR和Western Blot技术分别测定氧化应激通路和NLRP3炎性小体的表达水平。结果 表观特征观察显示,模型组小鼠体质量、腹部脂肪增多,肾周脂肪指数升高(P<0.01);给药后肾周脂肪指数明显下降(P<0.05,P<0.01)。YT+YR高剂量组,能降低血清总胆固醇(TC)、甘油三酯(TG)、葡萄糖(GLU)和低密度脂蛋白胆固醇(LDL-C)、谷丙转氨酶(ALT)、谷草转氨酶(AST)、丙二醛(MDA)水平,升高高密度脂蛋白胆固醇(HDL-C)、超氧化物歧化酶(SOD)水平,改善MAFLD组织病理学评分,降低肝脏指数及血清TNF-α、IL-1β、IL-18水平,升高Nrf2和HO-1的mRNA和蛋白表达水平,升高NQO-1的mRNA,降低NLRP3、ASC、Caspase-1、IL-1β及IL-18的mRNA和蛋白表达水平,差异有统计学意义(P<0.05,P<0.01)。结论 YT、YR可通过抑制NLRP3炎症小体的激活,增强Nrf2信号通路,调控氧化应激反应,从而改善高脂饮食诱导的MAFLD肝脏脂质沉积和炎症浸润,且YT+YR联合应用较单用更具优势。

Objective To investigate the improvement and molecular mechanism of hemp seed oil and algal oil mixture(YT) combined with Salviae Miltiorrhizae Radix et Rhizoma-Crtaegi Fructus-Notoginseng Radix et Rhizoma(YR)for treating metabolic associated fatty liver disease(MAFLD). Methods In present study,male C57BL/6J mice were fed with chow or a high fat diet(HFD)along with vehicle for 16 consecutive weeks,then intragastrically treated with YT,YR,high-and low-doses of YT+YR for the next 10 weeks. Serum levels of lipid accumulation,liver function, inflammatory factors and oxidative stress were detected. The degree of liver tissue lesions was analyzed using histological techniques, and protein levels involved in oxidative stress and NLRP3 inflammasome were measured by RT-PCR and Western Blot. Results Results of apparent characteristics showed that compared with the control group,the body mass,abdominal fat and the perirenal fat coefficient in the model group increased(P<0.01),the perirenal fat coefficient decreased significantly after administration(P<0.05,P<0.01). High-dose of YT + YR joint intervention could decrease the levels of serum total cholesterol(TC), triglyceride(TG), glucose(GLU),low-density lipoprotein cholesterol(LDL-C),alanine aminotransferase(ALT)and aspartate aminotransferase(AST),increase high-density of lipoprotein cholesterol(HDL-C),malondialdehyde(MDA)and superoxide dismutase(SOD), as well as improve MAFLD histopathological score(P<0.05);it was also found that liver coefficient,serum levels of TNF-α,IL-1β and IL-18 were decreased;the mRNA and protein expressions of Nrf2 and HO-1as well as the mRNA level of NQO-1 were increased;the mRNA and protein expression levels of NLRP3,ASC,Caspase-1,IL-1β and IL-18 were decreased,and the difference was statistically significant(P<0.05,P<0.01).Conclusion It has been demonstrated that YT,YR could improve liver lipid deposition and inflammation infiltration of HFD-induced MAFLD by inhibiting NLRP3 inflammasome activation,strengthening Nrf2 signaling pathway and regulation of oxidative stress. In general,YT and YR combined treatment for MAFLD has obvious advantages than the single treatment.