目的 通過(guò)網(wǎng)絡(luò)藥理學(xué)以及動(dòng)物實(shí)驗(yàn)探討六棱菊對(duì)代謝相關(guān)脂肪性肝?。∕AFLD）模型小鼠的干預(yù)作用及潛在的分子機(jī)制。方法 通過(guò)文獻(xiàn)檢索得到六棱菊的活性成分，通過(guò)SwissTargetPredicton和疾病數(shù)據(jù)庫(kù)（GeneCards）和PharmMapper數(shù)據(jù)庫(kù)預(yù)測(cè)六棱菊成分相關(guān)靶標(biāo)，Genecards數(shù)據(jù)庫(kù)獲取MAFLD和氧化應(yīng)激相關(guān)靶標(biāo)，通過(guò)STRING和Cytoscape3.9.1軟件建立蛋白質(zhì)-蛋白質(zhì)相互作用（PPI）網(wǎng)絡(luò)模型并找到關(guān)鍵靶點(diǎn)。對(duì)六棱菊治療MAFLD的靶標(biāo)進(jìn)行GO富集分析與KEGG通路富集分析。將50只C57BL/6J雄性無(wú)菌小鼠適應(yīng)性飼養(yǎng)1周后隨機(jī)分為5組，分別為對(duì)照組、模型組及六棱菊提取物低、中、高劑量（1.25、2.50、5.00 g·kg^-1）組；小鼠給予高脂飼料8周進(jìn)行MAFLD造模，造模后給藥6周。實(shí)驗(yàn)第14周末處死小鼠，比較各組小鼠肝臟指數(shù)、肝組織病理變化，測(cè)定各組小鼠血清中炎癥因子的水平及肝臟中脂肪含量，測(cè)定血清轉(zhuǎn)氨酶、肝臟脂肪水平以及肝組織的氧化損傷指標(biāo)丙二醛（MDA）、還原型谷胱甘肽（GSH）、超氧化物歧化酶（SOD）水平，檢測(cè)核因子E₂相關(guān)因子2 （Nrf2）信號(hào)通路重要基因和蛋白Nrf2、NAD（P）H喹啉氧化酶1 （Nqo1）、糖原合成酶激酶-3β （GSK-3β）和非受體酪氨酸蛋白激酶（Fyn）的表達(dá)水平。結(jié)果 網(wǎng)絡(luò)藥理學(xué)獲得六棱菊、MAFLD、氧化應(yīng)激的交集靶點(diǎn)共304個(gè)，六棱菊可能通過(guò)TNF、AKT1、ALB、EGFR、STAT3、NFE₂L2等核心靶點(diǎn)作用于HIF-1、糖尿病并發(fā)癥中的AGE-RAGE、甲狀腺激素、FoxO、PI3K-Akt、TNF信號(hào)通路等發(fā)揮MAFLD治療作用。動(dòng)物實(shí)驗(yàn)結(jié)果顯示，與對(duì)照組比較，模型組小鼠的整體狀態(tài)差，肝臟組織大面積脂肪變性及脂質(zhì)浸潤(rùn)，血清轉(zhuǎn)氨酶、肝臟脂肪水平明顯升高，證實(shí)了高脂飼料飲食喂養(yǎng)8周成功構(gòu)建了小鼠MAFLD模型；各藥物干預(yù)治療組小鼠肝臟指數(shù)、肝臟病理變化和肝臟脂肪水平，血清轉(zhuǎn)氨酶、脂質(zhì)過(guò)氧化水平和Nrf2信號(hào)通路重要基因和蛋白表達(dá)均較模型組顯著改變（P＜0.05），其中，六棱菊提取物高劑量組改變最為明顯。結(jié)論 不同劑量六棱菊提取物干預(yù)均能明顯改善MAFLD小鼠血清中炎癥因子水平，肝臟脂質(zhì)沉積、肝功能指標(biāo)，降低肝臟指數(shù)，可能與調(diào)控Nrf2信號(hào)通路的表達(dá)，降低肝組織中脂質(zhì)過(guò)氧化水平和炎癥反應(yīng)的表達(dá)有關(guān)。

Objective To investigate the effect and potential molecular mechanism of Laggera alata extract on alleviates metabolismrelated fatty liver disease (MAFLD) through network pharmacology and drug intervention. Methods The active ingredients of Laggera alata extract were analyzed through literature retrieval and the elated targets were predicted by SwissTargetPredicton and Pharm Mapper databases. MAFLD and oxidative stress-related targets were obtained by Genecards database. PPI network model was established and key targets were found by STRING and Cytoscape 3.9.1 software. Besides, GO enrichment analysis and KEGG pathway enrichment analysis were performed on the targets of hexagon chrysanthemum in the treatment of MAFLD. 50 male sterile C57BL/6J mice were randomly divided into five groups after one week of adaptive feeding, which were control group, model group, low dose group, medium dose group and high dose group of Laggera alata extract. The experimental mice were given high-fat diet for MAFLD modeling, and the low, medium and high dose groups (1.25, 2.50, and 5.00 g·kg^-1) were given daily by gavage. Mice in each intervention treatment group were given ig administration once a day for six weeks. The mice were killed at the end of the 14th week of the experiment, and the liver index, liver tissue pathological changes, serum inflammatory factors and liver fat content, serum aminotransferase, liver fat level, oxidative damage index of liver malondialdehyde (MDA), reduced glutathione (GSH) and superoxide dismutase (SOD) levels and important genes and proteins of Nrf2 signaling pathway of the five groups of mice were compared. Expression levels of nuclear factor E₂ associated factor 2 (Nrf2), NAD(P)H quinoline oxidase 1 (Nqo1), glycogen synthase kinase-3β (GSK-3β) and non-receptor tyrosine protein kinase (Fyn). Results A total of 304 intersection targets of Laggera alata, MAFLD and oxidative stress were obtained by network pharmacology. Laggera alata may act on HIF-1, AGE-RAGE, thyroid hormone, FoxO, PI3K-Akt and TNF signaling pathways in diabetic complications through core targets such as TNF, AKT1, ALB, EGFR, STAT3 and NFE₂L2 to exert the therapeutic effect of MAFLD. Compared with the control group, the overall state of mice was poor, the liver tissues were large steatosis and lipid infiltration, and the serum aminotransferase and liver fat levels were significantly increased, which confirmed that the MAFLD model of mice was successfully constructed after eight weeks of high-fat diet. Liver index, liver pathological changes, liver fat level, serum aminotransferase, lipid peroxidation level and Nrf2 signaling pathway important gene and protein expression of mice in all drug intervention groups were significantly changed compared with model group, among which the changes were most obvious in high-dose group, and the results were statistically significant (P < 0.05). Conclusion Different doses of Laggera alata extract can significantly improve the levels of serum inflammatory factors, liver lipid deposition, liver function, and decrease liver index in MAFLD mice, which may be related to regulating the expression of Nrf2 signaling pathway, reducing the level of lipid peroxidation and the expression of inflammation in liver tissues.

R285.5

廣西自然科學(xué)基金青年基金項(xiàng)目（2024GXNSFBA010302）;廣西重點(diǎn)研發(fā)計(jì)劃項(xiàng)目（桂科AB21196057）;國(guó)家中醫(yī)藥管理局高水平中醫(yī)藥重點(diǎn)學(xué)科建設(shè)項(xiàng)目-少數(shù)民族藥學(xué)（壯藥學(xué)）［zyyzdxk-2023165］;廣西國(guó)際壯醫(yī)醫(yī)院”青苗工程“人才培育項(xiàng)目（編號(hào)：2022001）;廣西國(guó)際壯醫(yī)醫(yī)院培育項(xiàng)目（2023GZYJKT008）;廣西中醫(yī)藥多學(xué)科交叉創(chuàng)新團(tuán)隊(duì)項(xiàng)目（GZKJ2309）;廣西中醫(yī)藥大學(xué)“高層次人才培育創(chuàng)新團(tuán)隊(duì)”項(xiàng)目（2022A008）;廣西國(guó)際壯醫(yī)醫(yī)院2023年高層次人才隊(duì)伍建設(shè)三年行動(dòng)計(jì)劃項(xiàng)目（GZCX20231203）;廣西中醫(yī)藥大學(xué)第三批“岐黃工程”高層次人才團(tuán)隊(duì)培育項(xiàng)目（202414）