目的 基于“肠-肝轴”学说，从“肝保护”及“肠屏障”角度表征蓬莪术醋制后对肝、肠的调节效应，揭示蓬莪术醋制增效机制。方法 将昆明小鼠随机分为对照组、模型组、阳性药（0.06 g/kg）组及蓬莪术生、醋品水煎液高、中、低剂量（3、2、1 g/kg）组，各组ig给药（10 mL/kg），1次/d，连续给药4 d。末次给药1 h后，除对照组外，其余各组小鼠ip硫代乙酰胺（thioacetamide，TAA，100 mg/kg）建立肝损伤模型。测定小鼠肝脏指数；采用试剂盒检测血清中天冬氨酸氨基转移酶（aspartate aminotransferase，AST）、丙氨酸氨基转移酶（alanine aminotransferase，ALT）、总超氧化物歧化酶（superoxide dismutase，SOD）、丙二醛（malondialdehyde，MDA）、脂多糖（lipopolysaccharide，LPS）水平，检测肝脏组织肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-6（interleukin-6，IL-6）、IL-1β、IL-10以及肠道IL-17A和分泌型免疫球蛋白A（secretory immunoglobulin A，sIgA）的含量；苏木素-伊红（hematoxylin-eosin，HE）染色观察肝脏及回肠组织形态；免疫组织化学法检测回肠组织紧密连接蛋白1（zonula occludens proteins-1，ZO-1）、咬合蛋白（occludin）的表达；16S rDNA基因测序分析小鼠肠道菌群变化；Western blotting检测Toll样受体4（Toll-like receptor 4，TLR4）、髓样分化因子88（myeloid differentiation factor 88，MYD88）及核因子-κB-p65（nuclear factor kappa-B-p65，NF-κB-p65）蛋白的表达。结果 蓬莪术醋制前后均可改善肝损伤小鼠的肝脏炎症细胞浸润、肝细胞肿胀变大等变化，且醋制后显著降低肝脏指数、ALT、AST及MDA水平（P＜0.05、0.001），显著升高SOD水平（P＜0.001）；蓬莪术醋制前后均可缓解肝损伤小鼠回肠绒毛萎缩、上皮细胞紧密连接间隙增宽等变化，且醋制后显著升高occludin的蛋白表达（P＜0.05），显著逆转由肝损伤引起的厚壁菌门丰度升高（P＜0.05）、拟杆菌门丰度的降低（P＜0.05）以及厚壁菌门与拟杆菌门比值的升高（P＜0.05）；蓬莪术醋制前后均可升高肝损伤小鼠回肠内IL-17A（P＜0.01）和sIgA水平（P＜0.001），降低TNF-α、IL-1β、IL-6水平（P＜0.05），同时抑制TLR4/MyD88/NF-κB通路上关键蛋白MyD88、NF-κB-p65和TLR4（P＜0.05）的表达，且醋制后显著升高IL-10水平（P＜0.05），降低LPS水平（P＜0.01），抑制TLR4（P＜0.05）蛋白表达。结论 蓬莪术醋制后通过保护肠道机械屏障、调节肠道生物屏障、修复肠道免疫屏障，抑制“肠-肝轴”TLR4/MyD88/NF-κB通路，可更好地发挥肝保护作用、改善肝损伤。

Objective Based on “gut-liver axis” theory, the regulatory effects of raw and vinegar-processed Curcuma phaeocaulis on liver and intestine were studied from the perspectives of “liver protective effect” and “intestinal barrier effect”, in order to study the processing synergistic mechanism of vinegar-processed C. phaeocaulis leading into liver. Method Kunming mice were randomly divided into the control group, model group, positive group (0.06 g/kg), high-dose, medium-dose and low-dose groups (3, 2, 1 g/kg) of raw and vinegar-processed C. phaeocaulis decoction, with 10 mice in each group. Each group was administered intragastrically once a day for four consecutive days (10 mL/kg). One hour after administration on the 4th day, except for the control group, all groups were intraperitoneally injected with thioacetamide (TAA) at a dose of 100 mg/kg to establish the liver injury model. The liver wet weight and liver index of mice were detected. The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), superoxide dismutase (SOD), malondialdehyde (MDA), lipopolysaccharide (LPS) and the contents of tumor necrosis factor-α (TNF-α), interleukin-6(IL-6), IL-1β, IL-10, IL-17A and secretory immunoglobulin A (sIgA) were detected by biochemical kits. The morphology of liver and ileum tissues was observed by hematoxylin-eosin (HE) staining. The expressions of zonula occludens proteins-1 (ZO-1) and occludin in ileum tissues were detected by immunohistochemistry. 16S rDNA gene sequencing was used to analyze the changes of intestinal flora in mice. The expressions of Toll-like receptor4 (TLR4), myeloid differentiation factor 88 (MyD88) and nuclear factor kappa-B-p65 (NF-κB-p65) proteins were detected by Western blotting. Results After treatment with C. phaeocaulis, the liver inflammatory cell infiltration and swelling of liver cells were improved, and the liver index, ALT, AST and MDA were significantly decreased (P < 0.05, 0.001) while SOD was significantly increased (P < 0.001) after treatment with vinegar-processed C. phaeocaulis. After treatment with C. phaeocaulis, the changes of ileum villus atrophy and tight junction gap widening of epithelial cells were alleviated in mice with liver injury, and the expression of occludin (P < 0.05) was significantly increased after treatment with vinegar-processed C. phaeocaulis. 16S rDNA sequencing analysis showed that vinegar-processed C. phaeocaulis could significantly reverse the increase of Firmicutes abundance (P < 0.05), the decrease of Bacteroidetes abundance (P < 0.05) and the increase of Firmicutes/Bacteroidetes ratio (P < 0.05) caused by liver injury. The levels of IL-17A and sIgA were increased (P < 0.01, 0.001), and the levels of TNF-α, IL-1β and IL-6 were decreased (P < 0.05). The expression of key proteins MyD88, NF-κB-p65 and TLR4 (P < 0.05) in the TLR4/MyD88/NF-κB pathway were inhibited to varying degrees in all drug administration groups. In vinegar-processed C. phaeocaulis groups, the levels of IL-10 were increased (P < 0.05), the LPS levels were significantly decreased (P < 0.01), and the TLR4 protein expression was significantly inhibited (P < 0.05). Conclusion The vinegar-processing C. phaeocaulis can play a better role in liver protection. It can regulate the three intestinal barriers by inhibiting the key pathway TLR4/MyD88/NF-κB of “gut-liver axis”, so as to play the synergistic effect of vinegar-processing C. phaeocaulis leading into liver to alleviate liver injury based on “gut-liver axis”.

R285.5

国家自然科学基金面上项目(82274099);四川省科技厅面上项目(24NSFSC1101);成都市科学技术局2025年第一批重点研发项目(2024-YF05-01999-SN)