天枢、内关穴分别电针预处理对急性心肌梗死小鼠棕色脂肪中UCP1/BMP3b信号通路的影响

The Impact of Electroacupuncture Preconditioning at Tianshu (ST25) or Neiguan (PC6) Acupoints on the UCP1/BMP3b Signaling Pathway in Brown Adipose Tissue of Mice with Acute Myocardial Infarction

  • 摘要:
    目的 观察天枢穴和内关穴分别电针预处理对急性心肌梗死(AMI)小鼠心功能以及棕色脂肪组织(BAT)中UCP1/BMP3b信号通路的影响,探讨不同穴位电针改善心肌梗死的潜在机制。
    方法 以健康成年小鼠和BAT切除小鼠为研究对象,并将其对应分为假手术组、模型组、内关组、天枢组。在适应性喂养1周后,干预组分别进行20 min的双侧内关穴或天枢穴电针预处理,结扎冠状动脉左前降支建立AMI模型。BAT切除组在电针前进行肩胛部BAT切除,其余干预与前保持一致。采用超声心动图检测各组小鼠心功能变化;TTC染色观察各组小鼠心肌梗死面积;ELISA检测各组小鼠血清中cTnT和BMP3b水平;qPCR法检测小鼠BAT中β3-AR、UCP1、BMP3b mRNA的相对表达;Western blot检测小鼠BAT中BMP3b和心脏中p-Smad1/5蛋白的相对表达。
    结果 与假手术组相比,模型组小鼠的左心室EF、FS降低(P<0.001),白色梗死区域面积增大(P<0.001),血清中cTnT水平升高(P<0.001),BAT中β3-AR、UCP1、BMP3b的mRNA表达水平升高(P<0.05,P<0.01),BMP3b的蛋白表达升高(P<0.01)。血清中的BMP3b含量升高(P<0.001),心脏中p-Smad1/5蛋白表达升高(P<0.01)。与模型组相比,天枢组和内关组小鼠的左心室EF、FS均升高(P < 0.001),白色梗死区域面积减小(P<0.001),血清中cTnT水平下降(P<0.05,P<0.01);天枢组BAT中β3-AR、UCP1、BMP3b的mRNA相对表达量明显升高(P <0.05,P<0.01,P<0.001),BMP3b的蛋白表达升高(P<0.01),心脏中p-Smad1/5蛋白表达升高(P<0.01),而内关组无明显变化。在BAT切除后,与模型组相比,内关组小鼠的左心室EF、FS升高(P<0.001),白色梗死区域面积减小(P< 0.001),血清中cTnT下降(P<0.001),而天枢组无明显变化;内关组和天枢组心脏中p-Smad1/5的蛋白表达和血清中BMP3b含量均无明显变化。
    结论 天枢穴或内关穴电针预处理都可以产生心肌保护效应,天枢穴的保护效应可能通过影响棕色脂肪中UCP1/BMP3b信号通路产生,而内关穴不依赖该通路。

     

    Abstract:
    OBJECTIVE To observe the effect of electroacupuncture (EA) preconditioning at Tianshu (ST25) and Neiguan (PC6) on cardiac function and UCP1/BMP3b signaling pathway in brown adipose tissue (BAT) in mice with acute myocardial infarction (AMI), so as to explore the potential mechanism of EA at different acupoints in improving myocardial infarction.
    METHODS Healthy adult mice and BAT excision mice were selected as the research objects, and they were divided into sham operation group, model group, PC6 group, and ST25 group. After one week of adaptive feeding, the mice in the intervention group were pretreated with bilateral EA at PC6 or ST25 for 20 minutes, respectively, and the AMI model was established by ligating the left anterior descending branch of the coronary artery. In the BAT resection group, BAT resection of the scapular region was performed before EA, and the rest of the intervention remained the same as before. Echocardiography was used to detect the changes in cardiac function. TTC staining was used to observe the myocardial infarct size. ELISA was used to detect the serum levels of cTnT and BMP3b in each group. The qPCR was used to detect the relative expression of β3-AR, UCP1 and BMP3b mRNA in mouse BAT. The protein expression of BMP3b in BAT and p-Smad1/5 in the heart were detected by Western blot.
    RESULTS Compared with the sham operation group, the left ventricular EF and FS of the model group mice were decreased (P<0.001), the white infarct area was increased (P<0.001), the cTnT level in serum was increased (P<0.001), the mRNA expression levels of β3-AR, UCP1, and BMP3b in BAT were increased (P<0.05, P<0.01), and the protein expression of BMP3b was increased (P<0.01). The BMP3b content in serum was increased (P<0.001), and the protein expression of p-Smad1/5 in the heart was increased (P<0.01). Compared with the model group, the left ventricular EF and FS of mice in the ST25 and PC6 group were increased (P<0.001), the area of white infarction was reduced (P<0.001), and the cTnT level in serum was decreased (P<0.05, P<0.01). The mRNA contents of β3-AR, UCP1, and BMP3b in BAT of the ST25 group were significantly increased (P<0.05, P<0.01, P<0.001), the protein expression of BMP3b was increased (P<0.01), and the protein expression of p-Smad1/5 in the heart was increased (P<0.01), while there was no significant change in the PC6 group. After BAT resection, compared with the model group, the left ventricular EF and FS of the mice in the PC6 group were increased (P<0.001), the area of white infarction was reduced (P<0.001), and the cTnT in serum was decreased (P<0.001), while there was no significant change in the ST25 group; there was no significant change in the protein expression of p-Smad1/5 in the heart and the BMP3b content in the serum of the PC6 and ST25 groups.
    CONCLUSION EA pretreatment at either ST25 or PC6 acupoints can produce myocardial protective effects. The protective effect of ST25 may be through influencing the UCP1/BMP3b signaling pathway in BAT, while PC6 does not depend on this pathway.

     

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