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目的探讨双酚A对大鼠背根神经节神经元钙离子通道各亚型的影响。方法选择50只健康4~6周龄清洁级SD大鼠,采用Ⅰ型胶原酶(4 mg/L)和Ⅰ型胰蛋白酶(1.5 mg/L)消化法分离背根神经节(DRG)神经元,以L、N、P/Q型钙通道阻断剂硝苯地平(nifedipine,10μmol/L)、乙酸齐考诺肽(ω-conotoxin MVIIA,2μmol/L)、ω-芋螺毒素(ω-contoxin MVIIC,2μmol/L)预孵育神经元20 min,特异性阻断通道电流,记录电流前即时通过灌流系统加入10μmol/L BPA,采用全细胞膜片钳方法记录钙电流。结果双酚A对各亚型通道均有抑制作用,其中,以L型钙通道为主。阻断L型钙通道,10μmol/L BPA对钙电流的抑制率由(53.22±11.00)%降低至(14.02±5.51)%,差异有统计学意义(P<0.05);BPA使L型钙通道电流密度(p A/p F)峰值由-30.54±3.92减小到-20.7±3.92,差异有统计学意义(P<0.05)。阻断N、P/Q型钙通道,对钙电流的抑制率由(53.22±11.00)%分别降低至(26.90±12.33)%和(25.66±5.99)%,差异有统计学意义(P<0.05)。结论双酚主要通过抑制L钙通道亚型,抑制钙离子电流。
Objective To investigate the effects of bisphenol A on calcium channels in rat dorsal root ganglion neurons. Methods Fifty healthy SD rats of 4 ~ 6 weeks of age were selected and dorsal root ganglion (DRG) neurons were isolated by digestion with type Ⅰ collagenase (4 mg / L) and type Ⅰ trypsin (1.5 mg / The effects of nifedipine (10μmol / L), ω-conotoxin MVIIA (2μmol / L), ω-conotoxin (ω- Contoxin MVIIC and 2μmol / L preincubated neurons for 20 min, blocking the channel current specifically. The current was immediately added to 10μmol / L BPA via perfusion system before recording current, and the whole-cell patch clamp method was used to record the calcium current. Results Bisphenol A had inhibitory effects on all the subtypes of channels, of which, L-type calcium channels were the most important. The inhibition of L-type calcium channel by 10μmol / L BPA decreased from (53.22 ± 11.00)% to (14.02 ± 5.51)%, the difference was statistically significant (P <0.05) The peak current density (p A / p F) decreased from -30.54 ± 3.92 to -20.7 ± 3.92, the difference was statistically significant (P <0.05). The inhibition of calcium channel in N, P / Q type calcium channel decreased from (53.22 ± 11.00)% to (26.90 ± 12.33)% and (25.66 ± 5.99)%, respectively, with statistical significance ). Conclusion Bisphenol mainly inhibits the calcium current by inhibiting the L calcium channel subtype.