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目的探讨心力衰竭(心衰)患者血清中的抗β3肾上腺素能受体(β3-AR)自身抗体的生物学效应,为临床治疗心衰提供新的思路和线索。方法(1)以人β3-AR细胞外第二环的合成肽段作为抗原,采用ELISA筛选正常人和心衰患者血清中抗β3-AR自身抗体。(2)提纯该抗体阳性的患者血清中的IgG。(3)设立各组对照观察该抗体对成年大鼠心肌细胞收缩效应的影响。(4)设立各组对照观察该抗体对乳鼠心肌细胞跳动频率的影响。结果(1)正常人抗β3-AR自身抗体的阳性率为11.0%,平均滴度为1∶14.59±1.61;心衰患者抗β3-AR自身抗体的阳性率为26.7%,平均滴度为1∶43.27±2.71;与正常人相比,P<0.05。(2)与空白对照组相比,该抗体可降低成年大鼠心肌细胞收缩幅度/初长度(3.84%±0.33%)、收缩速率(-0.47μm/s±0.07μm/s)和舒张速率(0.17μm/s±0.02μm/s),P<0.05。该作用不能被纳多洛尔(nadolol,β1-AR和β2-AR受体拮抗剂)阻断,但可被布拉洛尔(bupranolol,非特异性β受体拮抗剂)或β3抗原阻断。(3)与空白对照组(94.3次/min±10.7次/min)相比,该抗体可降低乳鼠心肌细胞跳动频率(47.1次/min±8.11次/min),P<0.05。同样该作用不能被纳多洛尔阻断,但可被布拉洛尔或β3抗原阻断。此外,该抗体的负性变时效应在观察时间内(6h)无衰减。结论心衰患者血清中含有较高滴度的抗β3-AR自身抗体并具有负性变力和变时效应,提示该抗体可能参与心衰的病理生理机制。
Objective To investigate the biological effects of anti-β3-adrenergic receptor (β3-AR) autoantibodies in the serum of patients with heart failure (HF), and to provide new ideas and clues for the clinical treatment of HF. Methods (1) The anti-β3-AR autoantibodies in the serum of normal and heart failure patients were screened by using the synthetic peptide of the second extracellular domain of human β3-AR as antigen. (2) Purify IgG in serum of the antibody-positive patient. (3) To establish the control group to observe the effect of this antibody on the contractile effect of cardiomyocytes in adult rats. (4) To establish the control group to observe the effect of the antibody on the beating frequency of neonatal rat cardiomyocytes. Results (1) The positive rate of anti-β3-AR autoantibodies in normal subjects was 11.0% with an average titer of 1:14.59 ± 1.61. The positive rate of anti-β3-AR autoantibodies in patients with heart failure was 26.7% and the average titer was 1 : 43.27 ± 2.71; P <0.05 compared with normal subjects. (2) Compared with the blank control group, the antibody could reduce the contractile rate / initial length (3.84% ± 0.33%), systolic velocity (-0.47μm / s ± 0.07μm / s) and diastolic velocity 0.17 μm / s ± 0.02 μm / s), P <0.05. This effect can not be blocked by nadolol (β1-AR and β2-AR receptor antagonists) but can be blocked by bupranolol or a β3 antigen. (3) Compared with the blank control group (94.3 times / min ± 10.7 times / min), the antibody can reduce the frequency of cardiomyocyte beating (47.1 times / min ± 8.11 times / min), P <0.05. Again this effect can not be blocked by nadolol, but it can be blocked by buraol or β3 antigen. In addition, the negative variable effect of the antibody did not attenuate within the observation time (6h). Conclusion The serum of patients with heart failure contains high titers of anti-β3-AR autoantibodies and has negative variable and time-varying effects, suggesting that the antibody may be involved in the pathophysiology of heart failure.