目的 通過網(wǎng)絡(luò)藥理學(xué)和動(dòng)物實(shí)驗(yàn)探究丹參-紅花藥對(duì)治療心肌缺血的潛在作用和機(jī)制。方法 檢索中藥系統(tǒng)藥理學(xué)數(shù)據(jù)庫與分析平臺(tái)（TCMSP）和文獻(xiàn)手動(dòng)補(bǔ)充確定丹參-紅花的活性成分及治療心肌缺血靶點(diǎn)。以“心肌缺血”及“急性心肌缺血”為關(guān)鍵詞，在CTD、DisGeNET、GeneCards和OMIM數(shù)據(jù)庫中檢索疾病的潛在靶點(diǎn)。利用Venny 2.1.0將篩選得到的有效化學(xué)成分靶點(diǎn)與疾病靶點(diǎn)進(jìn)行交集，確定丹參-紅花藥對(duì)治療心肌缺血的作用靶點(diǎn)。將交集靶點(diǎn)信息上傳至STRING v11.0，構(gòu)建活性成分-靶點(diǎn)網(wǎng)絡(luò)及蛋白質(zhì)相互作用網(wǎng)絡(luò)（PPI）。通過DAVID數(shù)據(jù)庫對(duì)丹參-紅花治療心肌缺血的靶點(diǎn)進(jìn)行基因本體（GO）功能和京都基因與基因組百科全書（KEGG）通路的富集分析。40只SD雄性大鼠，根據(jù)體質(zhì)量隨機(jī)分成4組，每組10只，即對(duì)照組、模型組、復(fù)方丹參滴丸（73 mg·kg^-1·d^-1）組、丹參-紅花（生藥量16 g·kg^-1·d^-1）組。對(duì)照組和模型組大鼠每天ig等量0.9%氯化鈉溶液，給藥組ig相應(yīng)劑量的藥物，每天1次，共7 d。于第6天，除對(duì)照組外，其余各組給藥1 h后sc異丙腎上腺素（ISO，85 mg·kg^-1），連續(xù)2 d造模。HE染色觀察各組大鼠心肌組織病理變化；蛋白免疫印跡法檢測各組大鼠心肌組織磷酸化哺乳動(dòng)物雷帕霉素靶蛋白（p-mTOR）、磷酸化轉(zhuǎn)錄激活蛋白3（p-STAT3）、缺氧誘導(dǎo)因子1α（HIF-1α）、血管內(nèi)皮生長因子A（VEGFA）、血小板衍生生長因子A（PDGFA）和堿性成纖維細(xì)胞生長因子（bFGF）蛋白表達(dá)水平。結(jié)果 網(wǎng)絡(luò)藥理學(xué)篩選得到丹參-紅花藥對(duì)治療心肌缺血的活性成分91個(gè)，靶點(diǎn)246個(gè)。PPI網(wǎng)絡(luò)結(jié)果顯示主要靶點(diǎn)包括STAT3、MAPK1、JUN、RELA、MAPK3、AKT1、SRC、APP、TNF、PIK3CA、CXCL8、KNG1、IL6、VEGFA、MAPK14、HSP90AA1、MAPK8、EGFR、FOS和IL2等。主要靶點(diǎn)相關(guān)的通路涉及血管生成（如HIF-1信號(hào)通路和VEGF信號(hào)通路）、炎癥反應(yīng)（如NF-κB信號(hào)通路和TNF信號(hào)通路）和細(xì)胞凋亡（如細(xì)胞凋亡信號(hào)通路和p53信號(hào)通路）等。動(dòng)物實(shí)驗(yàn)HE染色結(jié)果表明，對(duì)照組大鼠心肌細(xì)胞正常；模型組大鼠心肌纖維腫脹且有斷裂，心肌細(xì)胞增寬，局部有明顯水腫、滲出、炎性細(xì)胞浸潤；復(fù)方丹參滴丸組與模型組大鼠比較，心肌纖維排列略微紊亂且較窄，病理改變程度有所減輕。丹參-紅花組大鼠的心肌纖維排列整齊，細(xì)胞形態(tài)結(jié)構(gòu)完整，心肌纖維腫脹輕，無毛細(xì)血管擴(kuò)張，接近正常心肌形態(tài)。蛋白質(zhì)免疫印跡檢測結(jié)果表明，與對(duì)照組比較，模型組大鼠心肌組織中HIF-1α、VEGFA、PDGFA和bFGF的蛋白表達(dá)水平顯著升高（P＜0.05、0.01），p-mTOR和p-STAT3的蛋白表達(dá)水平顯著下降（P＜0.05、0.01）。與模型組比較，丹參-紅花顯著上調(diào)了p-mTOR、p-STAT3、HIF-1α、VEGFA、PDGFA和bFGF的蛋白表達(dá)水平（P＜0.05、0.01）。結(jié)論 丹參-紅花可能通過上調(diào)與血管生成密切相關(guān)的蛋白水平來促進(jìn)急性心肌缺血模型大鼠缺血心肌組織的血管生成，從而改善其缺血性損傷。

Objective To explore the potential effect and mechanism of Salvia miltiorrhiza-Carthamus tinctorius on myocardial ischemia through network pharmacology and animal experiments. Methods The pharmacological database and analysis platform (TCMSP) of the traditional Chinese medicine system and the literature were searched and manually supplemented to determine the active components of S. miltiorrhiza-C. tinctorius and the therapeutic target of myocardial ischemia. With "myocardial ischemia" and "acute myocardial ischemia" as the keywords, search the potential targets of diseases in CTD, DisGeNET, GeneCards and OMIM databases. Venny 2.1.0 was used to intersect the screened effective chemical component target with the disease target, and determine the target of S. miltiorrhiza-C. tinctorius for the treatment of myocardial ischemia. Upload the intersection target information to STRING v11.0 to build the active component-target network and protein-protein interaction (PPI) network. The DAVID database was used to enrich and analyze the Gene Ontology (GO) function and the Kyoto gene and genome encyclopedia (KEGG) pathway of the target of S. miltiorrhiza-C. tinctorius in the treatment of myocardial ischemia. Forty SD male rats were randomly divided into four groups according to their body mass, 10 in each group, namely, control group, model group, Compound Salvia Miltiorrhiza Dropping Pills (73 mg·kg^-1·d^-1) group, and S. miltiorrhiza-C. tinctorius (16 g·kg^-1·d^-1) group. The rats in the control group and model group were given the same amount of 0.9% sodium chloride solution once a day for seven days. On the 6th day, except for the control group, the rats in other groups were administered with isoproterenol (ISO, 85 mg·kg^-1) by sc for two days. HE staining was used to observe the pathological changes of myocardial tissue of rats in each group. Detection of p-mTOR, p-STAT3, HIF-1α, VEGFA, PDGFA and bFGF protein expression level in myocardial tissue of rats in each group by Western blotting. Results 91 active components and 246 targets of S.miltiorrhiza-C. tinctorius were screened by network pharmacology. PPI network results show that the main targets include STAT3, MAPK1, JUN, RELA, MAPK3, AKT1, SRC, APP, TNF, PIK3CA, CXCL8, KNG1, IL6, VEGFA, MAPK14, HSP90AA1, MAPK8, EGFR, FOS and IL2. The main target-related pathways involve angiogenesis (such as HIF-1 signal pathway and VEGF signal pathway), inflammatory response (such as NF-κB signal pathway and TNF signal pathway) and apoptosis (such as apoptosis signal pathway and p53 signal pathway). The results of HE staining in animal experiments showed that the myocardial cells in the control group were normal. In the model group, the myocardial fibers were swollen and the area was broken, the myocardial cells were widened, and there were obvious edema, exudation and inflammatory cell infiltration in the local area. Compared with the model group, the arrangement of myocardial fibers in the Compound Danshen Dropping Pills group was slightly disordered and narrow, and the degree of pathological changes was alleviated. The myocardial fibers of the rats in the S. miltiorrhiza-C. tinctorius group were arranged in order, the cell morphology and structure were complete, the myocardial fibers swelled slightly, and there was no telangiectasia, which was close to the normal myocardial morphology. The result of Western blotting showed that compared with the control group, the protein expression levels of HIF-1α, VEGFA, PDGFA and bFGF in the myocardial tissue of rats in the model group were significantly increased (P＜0.05, 0.01), while the protein expression levels of p-mTOR and p-STAT3 were significantly decreased (P＜0.05, 0.01). Compared with the model group, S. miltiorrhiza-C. tinctorius significantly up-regulated the protein expression level of p-mTOR, p-STAT3 and HIF-1α, VEGFA, PDGFA and bFGF (P＜0.05, 0.01). Conclusion S. miltiorrhiza-C. tinctorius may promote the angiogenesis of ischemic myocardium in rats with acute myocardial ischemia by up-regulating the protein level related to angiogenesis, thus improving the ischemic injury.

R285.5

國家自然科學(xué)基金資助項(xiàng)目（81974544）