目的 利用網(wǎng)絡(luò)藥理學(xué)方法研究地菍總黃酮抗2型糖尿病的作用機(jī)制，通過(guò)動(dòng)物實(shí)驗(yàn)對(duì)靶點(diǎn)進(jìn)行驗(yàn)證。方法 通過(guò)文獻(xiàn)檢索收集地菍總黃酮的化學(xué)成分，并在Swiss Target Prediction平臺(tái)進(jìn)行篩選；依據(jù)Swiss Target Prediction、GeneCards、TTD、DrugBank數(shù)據(jù)庫(kù)預(yù)測(cè)和篩選地菍總黃酮成分治療2型糖尿病的作用靶點(diǎn)；由Cytoscape 3.7.1構(gòu)建“成分–疾病–靶點(diǎn)”網(wǎng)絡(luò)；String數(shù)據(jù)庫(kù)構(gòu)建靶點(diǎn)蛋白相互作用（PPI）網(wǎng)絡(luò)；R語(yǔ)言對(duì)靶點(diǎn)基因進(jìn)行基因本體（GO）富集分析及京都基因和基因組百科全書（KEGG）通路富集分析。以高脂飲食誘導(dǎo)和ip鏈脲佐菌素（STZ）方法構(gòu)建2型糖尿病模型大鼠，將2型糖尿病模型造模成功的40只大鼠隨機(jī)分成模型組，二甲雙胍組（0.2 g/kg）組，地菍總黃酮（0.34、0.45、0.60 g/kg）組，每組8只，繼續(xù)提供高脂飼料喂養(yǎng)，另設(shè)對(duì)照組。各組每天ig相應(yīng)藥物，1次/d，對(duì)照組和模型組ig 0.5% CMC-Na溶液，連續(xù)給藥5周。記錄大鼠體質(zhì)量、飲水量和攝食量，檢測(cè)大鼠血糖及血脂的相關(guān)指標(biāo)，運(yùn)用qPCR法檢測(cè)地菍總黃酮對(duì)2型糖尿病大鼠肝組織中磷脂酰肌醇-3-激酶（PI3K）、蛋白激酶B（Akt）、腺苷酸活化蛋白激酶（AMPK）作用靶點(diǎn)的調(diào)控作用。Western blotting法檢測(cè)各組大鼠肝臟中Akt、AMPK、固醇調(diào)節(jié)元件結(jié)合蛋白-1C（SREBP-1C）的蛋白相對(duì)表達(dá)量。結(jié)果 從地菍總黃酮成分中篩選得到槲皮素、芹菜素、柚皮素等8種活性成分，作用于98個(gè)2型糖尿病靶點(diǎn)，GO功能與KEGG富集分析表明，地菍總黃酮抗糖尿病主要通過(guò)調(diào)節(jié)PI3K/Akt信號(hào)通路、AMPK相關(guān)信號(hào)通路等來(lái)發(fā)揮抗2型糖尿病的作用。動(dòng)物實(shí)驗(yàn)證實(shí)，地菍總黃酮可降低2型糖尿病大鼠的空腹血糖（FBG），抑制血脂升高，上調(diào)2型糖尿病大鼠肝臟PI3K、Akt、AMPK mRNA表達(dá)，顯著增加Akt、AMPK蛋白表達(dá)含量，降低SREBP-1C蛋白表達(dá)含量（P＜0.01、0.05），對(duì)糖、脂代謝紊亂均具有改善作用，具有良好的抗2型糖尿病作用。結(jié)論 驗(yàn)證了地菍總黃酮多成分、多靶點(diǎn)、多途徑治療2型糖尿病的特點(diǎn)，其抗糖尿病的作用機(jī)制可能與調(diào)節(jié)PI3K、Akt、AMPK、SREBP-1C靶點(diǎn)的表達(dá)水平，從而改善糖脂代謝紊亂有關(guān)，為后續(xù)更進(jìn)一步研究作用機(jī)制提供參考。

Objective To study the mechanism of anti type 2 diabetes by Melastoma dodecandrum flavonoid by using the method of network pharmacology, and to verify the related targets by animal experiments. Methods To collect the chemical constituents of M. dodecandrum flavonoids through literature search, and screen the target of M. dodecandrum in treatment of type 2 diabetes based on the Swiss Target Prediction, GeneCards, TTD, and DrugBank databases platform. To construct the “active ingredients - type 2 diabetes - potential target” by Cytoscape 3.7.1 software. The target protein interaction network was constructed through the String platform, GO enrichment analysis and KEGG metabolic pathway were analyzed through the R language analysis platform. To construct type 2 diabetes mellitus model rats high-fat diet induction and ip administered with streptozotocin method. 40 rats successfully modeled type 2 diabetes mellitus model were randomly divided into model group, metformin group (0.2 g/kg) group, and M. dodecandrum flavonoids (0.34, 0.45, 0.60 g/kg) groups, with 8 rats in each group. High-fat diet was continued and a control group was set up. Each group was given the corresponding drug intragaically once daily, the control group and model group were given 0.5% CMC-Na solution intragaically for 5 weeks. The body mass, water intake, and food intake of rats were recorded, and the related indexes of blood glucose and blood lipid of rats were detected. qPCR was used to detect the regulatory effects of M. dodecandrum flavonoids flavones on the targets of PI3K, Akt, and AMPK in liver tissue of type 2 diabetic rats. The protein expressions levels of Akt, AMPK, and SREBP-1c in liver tissues of rats in each group were detected by Western blotting. Results 8 Active ingredients such as quercetin, apigenin, naringin were screened from M. dodecandrum flavonoids and acted on 98 type 2 diabetes targets. GO function and KEGG enrichment analysis showed that M. dodecandrum flavonoids play an anti-diabetic role in type 2 diabetes mainly by regulating PI3K/Akt signaling pathway and AMPK related signaling pathway. Animal experiments confirmed that M. dodecandrum flavonoids can decrease FBG, inhibit the increase of blood lipid, up-regulate the mRNA expression of PI3K, Akt, and AMPK in liver of type 2 diabetic rats, and significantly increase the protein expression levels of Akt and AMPK, reduce the expression of SREBP-1C protein (P＜0.01, 0.05), can improve the metabolic disorders of sugar and lipid, and has a good anti-type 2 diabetes effect. Conclusion This study verified the characteristics of M. dodecandrum flavonoids in treatment of type 2 diabetes with multi-component, multi-target and multi-pathway. The anti-diabetic mechanism of of M. dodecandrum flavonoids may be related to the regulation of the expression levels of PI3K, Akt, AMPK, and SREBP-1C targets, thereby improving the disorder of glucose and lipid metabolism, providing a reference for further research on the mechanism of action.

R977

廣西中醫(yī)藥重點(diǎn)學(xué)科壯藥學(xué)（GZXK-Z-20-64）；廣西一流學(xué)科建設(shè)項(xiàng)目（桂教科研[2022]1號(hào)）；廣西壯族自治區(qū)科學(xué)技術(shù)廳壯瑤藥重點(diǎn)實(shí)驗(yàn)室項(xiàng)目（桂科基字[2014]32號(hào)）；廣西協(xié)同創(chuàng)新中心建設(shè)項(xiàng)目（桂教科研[2014]13號(hào)）；廣西壯族自治區(qū)民族藥資源與應(yīng)用工程研究中心資助項(xiàng)目（桂發(fā)改高技函[2020]2605號(hào)）；廣西科技基地和人才專項(xiàng)（桂科AD21238031）；廣西科技計(jì)劃項(xiàng)目（桂科AB21196016）；廣西中醫(yī)藥大學(xué)研究生教育創(chuàng)新計(jì)劃項(xiàng)目（YCSY2022008，YCSW2023387）；廣西中醫(yī)藥大學(xué)校級(jí)科研項(xiàng)目（2022MS013）；廣西壯瑤藥重點(diǎn)實(shí)驗(yàn)室立項(xiàng)課題（GXZYZZ2022-16）