目的 探究烏頭堿對心力衰竭模型大鼠心功能及心室重構的影響及對miR-150-5p的調控作用。方法 將SPF級SD雄性大鼠隨機分為假手術組、模型組、曲美他嗪組、烏頭堿組、烏頭堿＋antagomir-NC組、烏頭堿＋miR-150-5p antagomir組，每組15只。除假手術組外，其余組均利用結扎左前降支冠狀動脈法建立心力衰竭大鼠模型。測定各組大鼠心功能指標左室舒張末期內徑（LVEDD）、左室收縮末期內徑（LVESD）、左室射血分數（LVEF）；ELISA檢測各組大鼠心肌損傷指標心肌肌鈣蛋白I（CTnI）、腦鈉肽（BNP）和N末端B型腦鈉肽前體（NT-proBNP）的水平；測定各組大鼠心臟和左心室質量指數；Masson染色觀察各組大鼠心肌組織形態(tài)；TUNEL染色檢測各組大鼠心肌細胞凋亡率；RT-qRCR檢測各組大鼠心肌組織中miR-150-5p和細胞周期蛋白D2（CCND2）的表達；雙熒光素酶報告基因實驗驗證miR-150-5p與CCND2的靶向關系；Western blotting檢測心肌細胞中CCND2蛋白的表達。結果 與模型組相比，烏頭堿組大鼠心功能指標LVEDD、LVESD、心肌損傷指標CTnI、BNP和NT-proBNP水平、心臟質量指數、左心室質量指數、心肌纖維化區(qū)域、心肌細胞凋亡率和CCND2 mRNA水平均降低（P＜0.05），LVEF、miR-150-5p水平升高（P＜0.05）；使用miR-150-5p antagomir進行回補實驗，結果顯示，烏頭堿對心力衰竭大鼠心功能和心室重構的保護作用被逆轉，且CCND2 mRNA水平升高（P＜0.05）；與miR-150-5p mimic-NC組相比，miR-150-5p mimic組心肌細胞CCND2蛋白表達顯著降低（P＜0.05），且雙熒光素酶報告基因實驗驗證了miR-150-5p與CCND2之間存在靶向關系。結論 烏頭堿通過上調miR-150-5p表達對心力衰竭大鼠心功能和心室重構發(fā)揮保護作用。

Objective To investigate the impacts of aconitine on cardiac function and ventricular remodeling in heart failure modelrats, and its regulatory effect on miR-150-5p during this process. Methods SPF grade SD male rats were randomly grouped into sham surgery group, model group, trimetazidine group, aconitine group, aconitine + antagonimir NC group, and aconitine + miR-150-5p antagonimir group, with 15 rats in each group. Except for the sham surgery group, all other groups established heart failure rat models by ligating the left anterior descending coronary artery. The left ventricular end diastolic diameter (LVEDD), left ventricular end systolic diameter (LVESD), and left ventricular ejection fraction (LVEF) were measured for cardiac function indicators of rats in each group. ELISA was applied to detect the levels of myocardial injury indicators cardiac troponin I (CTnI), brain natriuretic peptide (BNP), and N-terminal B-type brain natriuretic peptide precursor (NT-proBNP) of rats in each group. Heart and left ventricular mass index of rats in each group were measured. Masson staining was applied to observe the myocardial tissue morphology of rats in each group, TUNEL staining was applied to detect the apoptosis rate of myocardial cells of rats in each group. RT-qRCR was applied to detect the expression of miR-150-5p and cyclin D2 (CCND2) in myocardial tissue of rats in each group. The double luciferase reporter gene experiment verified the targeting relationship between miR-150-5p and CCND2. Western blotting was used to detect the expression of CCND2 protein in myocardial cells. Results Compared with the model group, the cardiac function indicators LVEDD, LVESD, myocardial injury indicators CTnI, BNP, and NT proBNP levels, cardiac mass index, left ventricular mass index, myocardial fibrosis area, myocardial cell apoptosis rate and the level of CCND2 mRNA in the aconitine group decreased (P < 0.05). The levels of LVEF and miR-150-5p increased (P < 0.05). Supplementation experiments with miR-150-5p antagomir showed that the protective effects of aconitine on cardiac function and ventricular remodeling in rats with heart failure were reversed, and CCND2 mRNA levels were increased (P < 0.05). Compared with miR-150-5p mimic-NC group, the expression of CCND2 protein in myocardial cells in miR-150-5p mimic group was significantly decreased (P < 0.05), and the double luciferase reporter gene experiment verified that there was a targeted relationship between miR-150-5p and CCND2. Conclusion Aconitine plays a protective role in cardiac function and ventricular remodeling in rats with heart failure by up-regulating the expression of miR-150-5p.

R285；R286.2

河北省中醫(yī)藥管理局科研課題（2022442）