目的 探討青藤堿調(diào)控Toll樣受體4（TLR4）/NADPH氧化酶4（NOX4）信號(hào)通路對(duì)心房顫動(dòng)大鼠炎癥反應(yīng)及心肌纖維化的影響。方法 將SD大鼠隨機(jī)分為對(duì)照組、模型組、青藤堿（20、40、80 mg/kg）組、維拉帕米組及青藤堿＋Pam3Cys組，每組12只。對(duì)照組大鼠通過(guò)舌下iv等量的生理鹽水以代替心房顫動(dòng)誘導(dǎo)液；其余各組大鼠均通過(guò)舌下iv心房顫動(dòng)誘導(dǎo)液構(gòu)建心房顫動(dòng)大鼠模型，建模成功后，進(jìn)行給藥處理，1次/d，持續(xù)2周。彩色多普勒超聲儀檢測(cè)大鼠左室射血分?jǐn)?shù)（LVEF）、左室短軸縮短率（LVFS）的變化；ELISA法檢測(cè)大鼠血清中白細(xì)胞介素-6（IL-6）、腫瘤壞死因子-α（TNF-α）水平；檢測(cè)大鼠左心室質(zhì)量指數(shù)；蘇木精–伊紅（HE）染色檢測(cè)大鼠心肌組織病理變化；Masson染色檢測(cè)大鼠心肌組織纖維化程度；qRT-PCR檢測(cè)左心室心肌組織中轉(zhuǎn)化生長(zhǎng)因子-β1（TGF-β1）、膠原蛋白Ⅰ型（Collagen-Ⅰ）mRNA表達(dá)；Western blotting檢測(cè)大鼠左心室心肌組織中TLR4、NOX4蛋白表達(dá)。結(jié)果 與模型組比較，青藤堿各劑量組大鼠心肌組織病理?yè)p傷及心肌纖維化程度有所改善，LVEF、LVFS顯著升高，IL-6、TNF-α水平，TGF-β1和Collagen-Ⅰ mRNA表達(dá)及TLR4、NOX4蛋白表達(dá)降低（P＜0.05）。與青藤堿組比較，青藤堿＋Pam3Cys組大鼠心肌組織病理?yè)p傷及心肌纖維化加重，LVEF、LVFS顯著降低，IL-6、TNF-α水平，TGF-β1和Collagen-Ⅰ mRNA表達(dá)及TLR4、NOX4蛋白表達(dá)顯著升高（P＜0.05）。結(jié)論 青藤堿可能通過(guò)抑制TLR4/NOX4通路抑制心房顫動(dòng)大鼠炎癥反應(yīng)及心肌纖維化。

Objective To investigate the effect of sinomenine on inflammatory response and myocardial fibrosis in rats with atrial fibrillation by regulating the Toll-like receptor 4 (TLR4)/NADPH oxidase 4 (NOX4) signaling pathway. Methods SD rats were randomly grouped into control group, model group, sinomenine (20、40、80 mg/kg) group, verapamil group, and sinomenine + Pam3Cys group, with 12 rats in each group. Rats in control group was injected with an equal amount of physiological saline through the sublingual vein instead of atrial fibrillation inducing fluid, rats in other groups were used to construct atrial fibrillation rat models by injecting atrial fibrillation inducing fluid into the sublingual vein. After successful modeling, medication was administered once daily for 2 weeks. Color doppler ultrasound was applied to detect changes in left ventricular ejection fraction (LVEF) and left ventricular short axis shortening rate (LVFS) in rats. ELISA method was applied to detect the levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in rat serum. The left ventricular mass index of rats was detected, HE staining was applied to detect pathological changes in rat myocardial tissue, Masson staining was applied to detect the degree of myocardial fibrosis in rats, qRT-PCR was applied to detect the expression of transforming growth factor-β1 (TGF-β1) and Collagen-I mRNA in left ventricular myocardial tissue. Western blotting was applied to detect the expression of TLR4 and NOX4 proteins in rat left ventricular myocardial tissue. Results Compared with the model group, the myocardial tissue pathological damage and severe myocardial fibrosis of rats in sinomenine group were improved, LVEF and LVFS were increased, the IL-6 and TNF-α levels, TGF-β1 and Collagen-Ⅰ mRNA expression, and TLR4, NOX4 protein expression were reduced (P < 0.05). Compared with sinomenine group, the myocardial tissue pathological damage and myocardial fibrosis of rats in sinomenine + Pam3Cys group were aggravated, LVEF and LVFS were reduced, IL-6 and TNF-α levels, TGF-β1 and Collagen-Ⅰ mRNA expression, and TLR4, NOX4 protein expression were increased (P < 0.05). Conclusion Sinomenine may inhibit inflammatory response and myocardial fibrosis in rats with atrial fibrillation by inhibiting the TLR4/NOX4 pathway.

R285；R286.2

山東省中醫(yī)藥科技發(fā)展計(jì)劃項(xiàng)目（2019-0034）