目的 探討牛舌草水提物（AIAE）抗心力衰竭作用及作用機制。方法 以AB野生型及Tg（cmlc2：EGFP）斑馬魚為研究對象，設置對照組、模型（200 μmol∙L⁻¹鹽酸維拉帕米）組、地高辛（陽性藥，10 μg·mL⁻¹）組、AIAE（0.1、0.5、2.5 mg·mL⁻¹）組。給藥后檢測各組斑馬魚心包面積、靜脈淤血面積、靜脈竇-動脈球（SV-BA）距離、心率、短軸縮短率（FS）、射血分數(shù)（LVEF）、血流速度變化；吖啶橙染色觀察心肌細胞凋亡，蘇木素-伊紅（HE）染色觀察心臟組織病理變化；試劑盒法檢測鈉尿肽系統(tǒng)中N末端腦鈉肽前體（NT-proBNP）含量；實時熒光定量PCR（qRT-PCR）法檢測腦鈉肽（bnp）及心鈉肽（anp）mRNA表達水平。通過轉錄組測序（RNA-seq）預測AIAE抗心力衰竭的相關信號通路，并利用qRT-PCR檢測信號通路相關基因的表達水平。結果 與模型組相比，AIAE組斑馬魚心包面積、靜脈淤血面積、SV-BA距離均顯著減?。?i>P＜0.05、0.01、0.001），心肌細胞凋亡數(shù)減少，心率、FS、LVEF、血流速度顯著升高（P＜0.05、0.01、0.001），心臟壁厚度有不同程度恢復，NT-proBNP含量、bnp、anp mRNA表達水平顯著降低（P＜0.05、0.01、0.001）。RNA-seq結果提示AIAE可能通過調控氧化磷酸化、糖酵解/糖異生等能量代謝相關通路發(fā)揮作用。qRT-PCR驗證結果表明AIAE可調控上述通路cox6a1、pck1等基因的異常表達（P＜0.05、0.01、0.001）。結論 AIAE通過調節(jié)氧化磷酸化、糖酵解/糖異生等信號通路，抑制鈉尿肽系統(tǒng)激活，抑制心肌細胞凋亡發(fā)揮抗心力衰竭作用。

Objective To investigate the effect and possible mechanism of aqueous extract of Anchusa italica (AIAE) on heart failure in zebrafish. Methods Zebrafish of AB wild type and Tg (cmlc2: EGFP) were used as research subjects. The control group, model group (200 μmol∙L⁻¹ verapamil hydrochloride), digoxin group (positive drug, 10 μg·mL⁻¹), and AIAE groups (0.1, 0.5, 2.5 mg·mL⁻¹) were set up. After administration, the pericardial area, venous congestion area, distance between sinus venosus and bulbus arteriosus (SV-BA), heart rate, fractional shortening (FS), left ventricular ejection fraction (LVEF), and blood flow velocity changes of zebrafish in each group were detected. Acridine orange staining was used to observe myocardial cell apoptosis, and hematoxylin-eosin (HE) staining was used to observe pathological changes in cardiac tissue. The content of N-terminal pro-brain natriuretic peptide (NT-proBNP) in the natriuretic peptide system was detected by kit method. The mRNA expression levels of brain natriuretic peptide (bnp) and atrial natriuretic peptide (anp) were detected by real-time fluorescence quantitative PCR (qRT-PCR). The related signaling pathways of AIAE in anti-heart failure were predicted by transcriptome sequencing (RNA-seq), and the expression levels of related genes in the signaling pathways were detected by qRT-PCR. Results Compared with the model group, the pericardial area, venous congestion area, SV-BA distance, NT-proBNP content, bnp and anp mRNA expression levels were significantly reduced (P < 0.05, 0.01, 0.001), the number of cardiomyocyte apoptosis was reduced, the heart rate, FS, LVEF and blood flow velocity were significantly increased (P < 0.05, 0.01, 0.001), and the heart wall thickness was restored to different degrees in the AIAE groups. RNA-seq results suggested that AIAE may play a role in regulating oxidative phosphorylation, glycolysis/gluconeogenesis and other energy metabolism-related pathways. qRT-PCR results showed that AIAE could regulate the abnormal expression of cox6a1, pck1 and other genes in this pathway (P < 0.05, 0.01, 0.001). Conclusion The AIAE plays an anti-heart failure role by regulating oxidative phosphorylation, glycolysis/gluconeogenesis and other signaling pathways, inhibiting the activation of NP system and inhibiting cardiomyocyte apoptosis.

R285.5

中央引導地方科技發(fā)展專項資金項目（ZYYD2024JD06）；天山創(chuàng)新團隊項目（2024D14018）；新疆維吾爾自治區(qū)重點實驗室開放課題（KLUMH-202404）