目的 观察高糖环境对大鼠背根神经节(DRG)细胞瞬时感受器电位离子通道香草素受体4(TRPV4)、蛋白激酶Cε(PK Cε)mRNA表达的影响,明确高糖环境对以上2种受体的调控作用.方法 急性分离新生大鼠DRG细胞,培养48 h后更换培养液并分组.按照培养基含糖量不同分为:对照组(5.5 mmol/L,C组)、高糖1组(17.5 mmol/L,H1组)、高糖2组(35.0 mmol/L,H2组).换液后48 h提取细胞总RNA并进行逆转录.用RT-PCR及实时定量PCR检测TRPV4和PKCε mRNA表达的变化.结果 与对照组比较,PKCε mRNA在H1组表达没有明显变化(P＞0.05),而在H2组表达显著上调(P＜0.05).与对照组比较,TRPV4 mRNA在H1组表达显著上调(P＜0.05),而在H2组表达没有明显变化(P＞0.05).结论 高糖环境可以上调TRPV4/PKCε基因的表达,但是2种基因明显上调时糖浓度不一致,说明TRPV4的激活除了PKCε的调控外还受其它因素调节.%Objective To observe the effects of high glucose environment on expressions of transient receptor potential vanilloid 4 (TR-PV4), protein kinase Cε (PKCε) mRNA in dorsal root ganglion (DRG) neurons, and to explore the regulation mechanisms of high glucose environment on both two receptors. Methods DRG neurons were isolated from the newborn rats and cultured for 48 h,then the medium was changed and the neurons were divided into 3 groups according to the different glucose concentration in culture medium: control group (5.5 mmol/L,C),high glucose group 1(17.5 mmol/L,H1)and high glucose group 2(35.0 mmol/L,H2). Total RNA of DRG neurons were extracted after 48 h for RT-PCR and quantitative real-time PCR analysis of TRPV4 and PKCe mRNA expressions. Results Compared to control group,PKCe mRNA expression of H1 was not significantly changed (P >0.05) but obviously up-regulated H2 (P < 0.05). TRPV4 mRNA expression in H1 was significantly up-regulated (P< 0.05) but not in H2 (P> 0.05). Conclusion High glucose environment can increase the expression of TRPV4 and PKCe mRNA; however, the expression of two genes were not absolutely dose-dependently with the glu-cose concentration level in culture medium. These findings indicated more complicated mechanisms to activate the TRPV4 except the regula-tion by PKC ε.
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