佛波酯对体外培养大鼠前庭上皮PKC表达的影响

Expression of PKC in vestibular epithelium of rat during organ culture in vitro

目的 观察体外培养条件下蛋白激酶 C(proteinkinase C,PKC)在佛波酯 (phogrbol 12 - myristate 13- acetate,PMA)诱导下大鼠前庭上皮的表达和分布特点 .方法 取新生大鼠椭圆囊 ,建立体外前庭器官培养模型 ,采用免疫组织细胞化学的方法 ,以 PKCβ,γ抗体为标记物 ,检测 PMA诱导后前庭上皮 PKCβ,γ的表达变化和分布特点 .结果 PKCβ,γ在正常培养新生大鼠前庭上皮细胞的细胞膜及细胞质中有弱表达 .PMA作用后 ,其表达逐渐增多 ,PKCβ1 主要位于细胞膜 ,在 30 min时表达最强 ,其后逐渐减少 ,4 5 min仍高于正常(P<0 .0 1) .PKCβ2 不但位于细胞膜 ,而且在纤毛亦明显表达 ;PKCγ在 PMA诱导后胞质表达增加 .在毛细胞层和支持细胞层均有 ,主要位于毛细胞层 .结论 体外前庭器官培养在 PMA诱导后 。

AIM To investigate the expression of PKCβ,γ of vestibular epithelium in rats after PMA treatment during organ culture in vitro so as to elucidate the biological character. METHODS The vestibular organ was placed in the bottom of a culture plate which contained culture medium to grow. After the initial culture period of 24 h, the explant was incubated with PMA for different periods of time. The expressions of PKCβ 1, β 2, and PKCγ were determined by anti PKCβ 1, β 2, and PKCγ with immunocytochemistry. RESULTS In the normal cultured vestibular epithelium of new born rats, the levels of expression of PKC were low in the control groups. In the experimental groups, PKC levels were elevated significantly and increased to the peak at 30 minutes. Then the levels decreased gradually, but at 45 minutes they were still increasing compared with the normal. PKC β staining was detected in cell membranes, PKCβ 2, not only stained in cell membranes but also in cilia, PKCγ staining increased in cell cytoplasm. CONCLUSION During organ culture in vitro , the expressions of PKCβ,γ of vestibular epithelium in rats after PMA treatment increased, which suggests that the isoform of PKC may have different forms of and different activation, isoform may function differently.