目的 基于“腸-肝軸”學(xué)說，從“肝保護(hù)”及“腸屏障”角度表征蓬莪術(shù)醋制后對肝、腸的調(diào)節(jié)效應(yīng)，揭示蓬莪術(shù)醋制增效機(jī)制。方法 將昆明小鼠隨機(jī)分為對照組、模型組、陽性藥（0.06 g/kg）組及蓬莪術(shù)生、醋品水煎液高、中、低劑量（3、2、1 g/kg）組，各組ig給藥（10 mL/kg），1次/d，連續(xù)給藥4 d。末次給藥1 h后，除對照組外，其余各組小鼠ip硫代乙酰胺（thioacetamide，TAA，100 mg/kg）建立肝損傷模型。測定小鼠肝臟指數(shù)；采用試劑盒檢測血清中天冬氨酸氨基轉(zhuǎn)移酶（aspartate aminotransferase，AST）、丙氨酸氨基轉(zhuǎn)移酶（alanine aminotransferase，ALT）、總超氧化物歧化酶（superoxide dismutase，SOD）、丙二醛（malondialdehyde，MDA）、脂多糖（lipopolysaccharide，LPS）水平，檢測肝臟組織腫瘤壞死因子-α（tumor necrosis factor-α，TNF-α）、白細(xì)胞介素-6（interleukin-6，IL-6）、IL-1β、IL-10以及腸道IL-17A和分泌型免疫球蛋白A（secretory immunoglobulin A，sIgA）的含量；蘇木素-伊紅（hematoxylin-eosin，HE）染色觀察肝臟及回腸組織形態(tài)；免疫組織化學(xué)法檢測回腸組織緊密連接蛋白1（zonula occludens proteins-1，ZO-1）、咬合蛋白（occludin）的表達(dá)；16S rDNA基因測序分析小鼠腸道菌群變化；Western blotting檢測Toll樣受體4（Toll-like receptor 4，TLR4）、髓樣分化因子88（myeloid differentiation factor 88，MYD88）及核因子-κB-p65（nuclear factor kappa-B-p65，NF-κB-p65）蛋白的表達(dá)。結(jié)果 蓬莪術(shù)醋制前后均可改善肝損傷小鼠的肝臟炎癥細(xì)胞浸潤、肝細(xì)胞腫脹變大等變化，且醋制后顯著降低肝臟指數(shù)、ALT、AST及MDA水平（P＜0.05、0.001），顯著升高SOD水平（P＜0.001）；蓬莪術(shù)醋制前后均可緩解肝損傷小鼠回腸絨毛萎縮、上皮細(xì)胞緊密連接間隙增寬等變化，且醋制后顯著升高occludin的蛋白表達(dá)（P＜0.05），顯著逆轉(zhuǎn)由肝損傷引起的厚壁菌門豐度升高（P＜0.05）、擬桿菌門豐度的降低（P＜0.05）以及厚壁菌門與擬桿菌門比值的升高（P＜0.05）；蓬莪術(shù)醋制前后均可升高肝損傷小鼠回腸內(nèi)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），同時(shí)抑制TLR4/MyD88/NF-κB通路上關(guān)鍵蛋白MyD88、NF-κB-p65和TLR4（P＜0.05）的表達(dá)，且醋制后顯著升高IL-10水平（P＜0.05），降低LPS水平（P＜0.01），抑制TLR4（P＜0.05）蛋白表達(dá)。結(jié)論 蓬莪術(shù)醋制后通過保護(hù)腸道機(jī)械屏障、調(diào)節(jié)腸道生物屏障、修復(fù)腸道免疫屏障，抑制“腸-肝軸”TLR4/MyD88/NF-κB通路，可更好地發(fā)揮肝保護(hù)作用、改善肝損傷。

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 “l(fā)iver 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

國家自然科學(xué)基金面上項(xiàng)目(82274099);四川省科技廳面上項(xiàng)目(24NSFSC1101);成都市科學(xué)技術(shù)局2025年第一批重點(diǎn)研發(fā)項(xiàng)目(2024-YF05-01999-SN)