论文部分内容阅读
电压门控钠通道广泛分布于各类细胞和组织中,参与许多生理功能的调节。作为位于脂质双分子层的膜蛋白,周围的质膜成分对于其门控特性和药理学特性是否存在影响仍然未知。本研究采用全细胞膜片钳技术,以两种钠通道的特异性调制剂BmK I和BmK AS为研究工具,在鞘磷脂酶D作用于细胞膜后,观察ND7-23细胞系上内源表达的电压门控钠通道的门控特性和药理学特性是否发生改变。结果显示,鞘磷脂酶D作用后,电压门控钠通道的门控特性并未发生变化,但其药理学特性发生了一定程度的改变。在低浓度30 nmol/L BmK I作用后,鞘磷脂酶D的修饰使得激活曲线的斜率因子k值发生改变,且30和100 nmol/L BmK I作用后,电压依赖性的慢失活和稳态失活发生超极化偏移。同样在低浓度0.1和10 nmol/L BmK AS作用后,鞘磷脂酶D的修饰使得电压依赖性的慢失活发生超极化偏移或斜率因子k值的改变。以上结果表明,通道毒理学依赖于周围的质膜环境。证明细胞膜可以调节钠通道的药理学特性。这不仅有助于对钠通道结构与周围膜蛋白相互作用关系的进一步理解,同时也为针对钠通道相关疾病的药物研发提供有益的参考思路。
Voltage-gated sodium channels are widely distributed in various types of cells and tissues and are involved in the regulation of many physiological functions. As membrane proteins located in the lipid bilayer, whether the surrounding plasma membrane components have an influence on their gating properties and pharmacological properties remains unknown. In this study, the whole-cell patch clamp technique was used to study the expression of endogenous voltage on ND7-23 cell line by using two kinds of sodium channel specific modulators BmK I and BmK AS as research tools after sphingomyelinase D acting on the cell membrane Gated sodium channel gating characteristics and pharmacological characteristics have changed. The results showed that after the action of sphingomyelinase D, the gating characteristics of voltage-gated sodium channel did not change, but its pharmacological properties changed to some extent. After 30 nmol / L BmK I at a low concentration, the kappa value of the slope of the activation curve was changed by the modification of sphingomyelinase D, and after 30 and 100 nmol / L BmK I treatment, the voltage-dependent slow inactivation and stabilization Hyperpolarization shift occurs in state inactivation. Also at low concentrations of 0.1 and 10 nmol / L BmK AS, the modification of sphingomyelinase D caused a shift in hyperpolarization or a change in the value of the slope factor, k, for a slow, voltage-dependent deactivation. The above results indicate that channel toxicology depends on the surrounding plasma membrane environment. Prove the cell membrane can regulate the pharmacological properties of sodium channels. This not only contributes to the further understanding of the relationship between the sodium channel structure and the surrounding membrane proteins, but also provides a useful reference for the drug development of sodium channel-related diseases.