Objectives: To explore the effect of resveratrol (Res) on Kawasaki disease (KD)-induced myocardial injury and to evaluate its effect on apoptosis and autophagy.
Methods: Forty-eight juvenile male Sprague Dawley rats were randomly divided into a control group, a Res group, a lactobacillus casei cell wall extract (LCWE)-induced Kawasaki disease group (KD group), and a LCWE-induced Kawasaki disease + Res treatment group (Res+KD group). The control group was intraperitoneally injected with saline. The Res group was intraperitoneally injected with resveratrol (100 mg/kg). The KD group was intraperitoneally injected with 0.5 mL LCWE (1 mg/mL). The Res+KD group was intraperitoneally injected with 0.5 mL LCWE (1 mg/mL) and resveratrol (100 mg/kg). After 4 weeks, the left ventricular ejection fraction (LVEF) and short axis shortening rate (LVFS) were detected by echocardiography. The apoptotic rate was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) staining. The levels of B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), microtubule-associated protein 1 light chain 3β (LC3B), Beclin-1, autophagy related 5 (Atg5) and sequestosome-1 (p62) were detected by Western blotting. The formation of autophagosome was observed under transmission electron microscope.
Results: There was no significant difference in the above-mentioned indexes between the control group and the Res group (all P>0.05). Compared with the control group, the values of LVEF and LVFS were significantly decreased in the KD group (both P<0.01); the ratio of Bax/Bcl-2 and TUNEL-positive cells were increased (both P<0.01); the LC3BII/LC3BI ratio, the levels of Beclin 1, Atg5 and p62, and the number of autophagosomes were also significantly increased in KD group (all P<0.01). Compared with the KD group, the values of LVEF and LVFS were significantly increased, the ratio of Bax/Bcl-2 and TUNEL-positive cells were decreased, the LC3BII/LC3BI ratio, the levels of Beclin 1, Atg5 and p62 were all decreased (all P<0.01), and the number of autophagosomes was suppressed.
Conclusions: Res can attenuate the KD-induced myocardial injury via inhibiting the apoptosis and autophagy.
目的: 探讨白藜芦醇(resveratrol，Res)在川崎病(Kawasaki disease，KD)诱发心肌损伤中的作用及其对细胞凋亡和自噬的调控作用。方法: 将48只幼年雄性Sprague Dawley大鼠(4周龄)随机分为对照组、Res对照组(Res组)、干酪乳酸杆菌细胞壁提取物(lactobacillus casei cell wall extract，LCWE)注射模拟KD组(KD组)以及LCWE注射模拟+Res处理组(Res+KD组)。对照组腹腔注射生理盐水，Res组腹腔注射Res(100 mg/kg)，KD组腹腔注射LCWE (1 mg/mL)，Res+KD组腹腔注射LCWE(1 mg/mL)和Res(100 mg/kg)。4周后，采用超声检测左室射血分数(left ventricular ejection fraction，LVEF)和短轴缩短率(left ventricular fraction shortening，LVFS)；采用原位末端标记法(terminal-deoxynucleoitidyl transferase mediated nick end labeling，TUNEL)染色观察细胞凋亡的变化；采用蛋白质印迹法检测心肌组织中B细胞淋巴瘤2(B-cell lymphoma-2，Bcl-2)、Bcl-2关联X蛋白(Bcl-2-associated X protein，Bax)、微管相关蛋白轻链3β(microtubule-associated protein 1 light chain 3β，LC3B)、自噬关键分子酵母Atg6同系物(Beclin 1)、自噬相关蛋白5(autophagy related 5，Atg5)和选择性自噬接头蛋白(sequestosone-1，p62)表达，并在透射电镜下观察自噬小体的形成。结果: 与对照组相比，Res组各项指标差异均无统计学意义(均P>0.05)；KD组LVEF和LVFS均明显降低(均P<0.01)，Bax/Bcl-2比值和TUNEL染色阳性细胞均明显增加(均P<0.01)，而LC3BII/LC3BI比值、Beclin 1、Atg5和p62表达也均显著增加(均P<0.01)，自噬小体增多；与KD组相比，Res+KD组LVEF和LVFS均增加(均P<0.01)，Bax/Bcl-2比值和TUNEL染色阳性细胞均减少(均P<0.01)，LC3BII/LC3BI比值、Beclin1、Atg5和p62表达也均显著降低(均P<0.01)，自噬小体的形成明显受到抑制。结论: Res能够通过抑制细胞凋亡和自噬减轻KD引起的心肌损伤。.
Keywords: Kawasaki disease; apoptosis; autophagy; resveratrol.