补骨脂素对溃疡性结肠炎小鼠肠道屏障及继发性肝损伤的改善作用  

Improvement effect of psoralen on intestinal barrier and secondary liver injury in mice with ulcerative colitis

作　　者：曹柳 唐晓晴 罗青 牛梦园 戴卫波[2] CAO Liu;TANG Xiao-qing;LUO Qing;NIU Meng-yuan;DAI Wei-bo(Zhongshan Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Traditional Chinese Medicine;Zhongshan Hospital of Traditional Chinese Medicine,Zhongshan 528400,China)

机构地区：[1]广州中医药大学附属中山中医院 [2]中山市中医院,中山528400

出　　处：《天然产物研究与开发》2024年第7期1121-1129,1148,共10页Natural Product Research and Development

基　　金：广东省自然科学基金(2023A1515011699);广东省医学科研基金(A2022479)。

摘　　要：研究补骨脂素(psoralen,PSO)对溃疡性结肠炎(ulcerative colitis,UC)小鼠的肠道屏障及继发性肝损伤的改善作用。40只雄性C57BL/6J小鼠随机分为对照组(control,Con)、模型组(model,Mod)、阳性药柳氮磺吡啶组(sulfasalazine,SASP,200 mg/kg)、PSO低剂量组(PSO-L,20 mg/kg)和PSO高剂量组(PSO-H,40 mg/kg),通过自由饮用2.25%的葡聚糖硫酸钠盐(dextran sulfate sodium salt,DSS)7 d构建UC模型。实验期间记录小鼠的体重,疾病活动指数(disease activity index,DAI)。实时荧光定量聚合酶链式反应(real-time quantitative polymerasechain reaction,RT-qPCR)检测结肠中带状闭合蛋白-1(zonula occludens-1,ZO1)、闭合蛋白-1(Claudin-1)和闭锁蛋白(Occludin)的表达。生化试剂盒检测肝脏中谷草转氨酶(aspartate aminotransferase,AST)、谷丙转氨酶(alanine transaminase,ALT)、碱性磷酸酶(alkaline phosphatase,AKP)和乳酸脱氢酶(lactate dehydrogenase,LDH)含量。酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测肝脏脂多糖(lipopolysaccharides,LPS)、C反应蛋白(C-reactiveprotein,CRP)、降钙素原(procalcitonin,PCT)、白细胞介素-6(interleukin-6,IL-6)以及肿瘤坏死因子-α(tumor necrosis factor,TNF-α)含量。蛋白印迹(Western blot)法检测肝脏Toll样受体4(Toll-like receptor 4,TLR4)、髓样分化因子88(myeloid differentiation primary response protein 88,MyD88)以及磷酸化核因子κB(phosphorylated nuclear factor kappa-B,p-NF-κB)的表达。结果显示,与Mod组相比,经PSO治疗后,UC小鼠体重下降得到缓解、DAI评分下降、缓解了结肠长度的缩短;ZO1、Claudin-1和Occludin的mRNA的表达均上调;肝脏AST、ALT、AKP和LDH含量均下降;肝脏LPS、CRP、PCT、IL-6以及TNF-α含量均下降;下调了肝脏TLR4、MyD88以及p-NF-κB蛋白的表达。以上结果表明,PSO可以改善DSS诱导的小鼠肠道屏障功能,并通过抑制TLR4/MyD88/NF-κB信号通路改善继发性肝损伤。This study aims to investigate the potential of psoralen(PSO)in ameliorating intestinal barrier dysfunction and secondary liver injury in mice with ulcerative colitis(UC).Forty male C57BL/6J mice were randomly assigned into the following groups:control group(Con),model group(Mod),positive control group treated with sulfasalazine(SASP,200 mg/kg),low-dose PSO group(PSO-L,20 mg/kg),and high-dose PSO group(PSO-H,40 mg/kg).Ulcerative colitis(UC)models were induced by freely drinking 2.25% dextran sulfate sodium salt(DSS)for seven days.During the experiment,the weight of mice and disease activity index(DAI)were recorded.Real-time quantitative polymerasechain reaction(RT-qPCR)was used to detect the expression of zonula occludens-1(ZO1),Claudin-1 and Occludin in the colon.Biochemical assay kits were employed to measure the levels of aspartate aminotransferase(AST),alanine transaminase(ALT),alkaline phosphatase(AKP)and lactate dehydrogenase(LDH)in the liver.Enzyme-linked immunosorbent assay(ELISA)was conducted to quantify the levels of lipopolysaccharides(LPS),C-reactive protein(CRP),procalcitonin(PCT),interleukin-6(IL-6)and tumor necrosis factor-alpha(TNF-α)in the liver.Western blot analysis was performed to assess the expression of Toll-like receptor 4(TLR4),myeloid differentiation primary response protein 88(MyD88)and phosphorylated nuclear factor kappa-B(p-NF-κB)in the liver.The results showed that compared to the Mod group,after PSO treatment,the weight loss of UC mice was alleviated,DAI scores decreased,and colonic length shortening was relieved.The mRNA expression of ZO1,Claudin-1,and Occludin was upregulated.The levels of AST,ALT,AKP and LDH in the liver decreased;the levels of LPS,CRP,PCT,IL-6 and TNF-α in the liver decreased;and the expression of TLR4,MyD88 and p-NF-κB proteins in the liver was downregulated.The above results indicate that PSO can improve intestinal barrier function in DSS-induced mice and ameliorate secondary liver injury by inhibiting the TLR4/MyD88/NF-κB signaling pathway.

关 键 词：补骨脂素 溃疡性结肠炎 肠道屏障 继发性肝损伤 炎症 

分 类 号：R285.5[医药卫生—中药学]