蛋白激酶C抑制剂对离体鼻息肉组织中嗜酸粒细胞凋亡的影响

The influence of protein kinase C inhibitor in eosinophil apoptosis of nasal polyps

目的 研究蛋白激酶C(proteinkinaseC ,PKC)抑制剂对鼻息肉组织嗜酸粒细胞凋亡的作用。方法 用细胞培养的方法 ,观察研究PKC抑制剂——— 1 (5 异喹啉硫酰氨 ) 3 甲基哌嗪 [1 (5 lsoquinolinyl sulfony) 3 methylpiperazinedihydrochloride ,Iso H7]对鼻息肉组织嗜酸粒细胞凋亡的影响 ,即将鼻息肉组织分为 2组 ,实验组在培养液中加不同浓度 (10 -6,10 -4,10 -2 mol/L)的抑制剂———Iso H7,分别在培养后的第 1、3、5天 ,各取出 1块 ,常规石蜡包埋 ,连续切片 ,用末端脱氧核苷酸转移酶介导的d UTP缺口末端标记法 (terminal deoxynucleotidyltransferased UTPnickendlabeling ,TUNEL)、凋亡原位杂交和MGG(May GrunwaldGiemsa)双染进行嗜酸粒细胞凋亡指数测定 ;对照组不加PKC抑制剂。结果 ①培养第 1天各浓度下的 2组标本均偶见凋亡的嗜酸粒细胞 ,培养到第 3天 ,实验组可见大量的嗜酸粒细胞凋亡 ,对照组第 1天与实验组第 1天比较嗜酸粒细胞凋亡指数差异无显著性 (t=1 72 0 0 ,P >0 0 5 ) ;②在实验组中 ,培养第 1天凋亡的嗜酸粒细胞与第 3天比较差异有显著性 (t=7 0 2 5 6 ,P <0 0 0 1) ,实验组第 3天与第 5天比较差异无显著性 (t=0 93995 ,P >0 0 5 ) ,但可见部分凋亡的嗜酸粒细胞膜破裂。

Objective To study the influence of the inhibitor of protein kinase C (PKC), Iso-H7 [1-(5-lsoquinolinyl-sulfony)-3-methylpiperazine dihydrochloride] on eosinophil apoptosis in nasal polyp tissues. Methods Nasal polyp tissues were cultured and divided into two groups, test group (with Iso-H7 in different concentration) and control group (without the inhibitor of PKC), to check out the influence of Iso-H7 on eosinophil apoptosis in nasal polyps. The nasal polyp tissues were cultured in Iso-H7 in concentration of 10 -6 ,10 -4 ,10 -2 mol/L for 1 d,3 d, and 5 d, respectively. Eosinophil apoptosis index was measured with TUNEL (terminal-deoxynucleotidyl transferase d-UTP nick end labeling), in situ hybridization and MGG staining to identify eosinophil apoptosis. Results Occasionally, there was little eosinophil apoptosis in two groups in 1 d,but in 3 d, There were a great deal of eosinophil apoptosis in test group. The eosinophil apoptosis indexs of the two groups were significantly different (P<0.01). The indexs of eosinophil apoptosis was not significantly higher in 3 d than that in 5 d in test groups (P>0.05),but significantly higher than that in 1d (P<0.01). Conclusions Iso-H7 takes a important role in accelerating eosinophil apoptosis. The inhibitor of PKC could significantly promote eosinophil apoptosis in nasal polyps. There is no dosage and time dependence of PKC inhibitor in regulating eosinophil apoptosis of nasal polyps.