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目的 探讨蛋白激酶C(PKC)信号转导途径在支气管哮喘 (简称哮喘 )大鼠气道平滑肌细胞 (ASMC)增殖中的作用。方法 (1) 4 8只Wistar大鼠分为哮喘组 (A组 )及对照组 (B组 ) ,根据激发时间 (2、4、8周 )又分别分为A1、A2 、A3 组和B1、B2 、B3 组 ,其中A、B组大鼠各 12只 ,A2 、A3 、B2 及B3 组各 6只。用流式细胞术、四甲基偶氮唑盐 (MTT)法、增殖细胞核抗原 (PCNA)染色等方法观察每组ASMC增殖 ;(2 )用PKC激活剂 12 肉蔻酰 13 乙酸佛波酯 (PMA)及抑制剂Ro 31 82 2 0分别干预A1、B1组ASMC ,观察ASMC增殖的变化 ;(3)用逆转录 聚合酶链测定 (RT PCR)和免疫细胞化学法检测A1、A2 、A3 组和B1组ASMCPKC α的表达。结果 (1)A组ASMCS期比例、吸光度 (A)值、PCNA表达增高 ,与B组比较差异有显著性 (P <0 0 1)。 (2 )A1组ASMCS期比例、A值、PCNA阳性表达率在干预前分别为 (19± 3) %、0 4 5 9± 0 0 36、(80± 10 ) % ;10nmol/LPMA处理后分别为 (2 7± 4 ) %、0 5 99± 0 0 78、(95± 9) % ;5 0nmol/LPMA处理后为 (14± 3) %、0 346± 0 0 38、(5 3± 8) % ;Ro 31 82 2 0处理后为 (14± 3) %、0 343± 0 0 4 8、(4 9± 8) %。各干预剂处理后与处理前比较差异均有显著性 (P <0 0 1) ;5 0nmol/LPMA处?
Objective To investigate the role of protein kinase C (PKC) signal transduction pathway in the proliferation of airway smooth muscle cells (ASMCs) in asthmatic rats. Methods (1) 48 Wistar rats were divided into asthma group (group A) and control group (group B), which were divided into groups A1, A2, A3 and B1 according to the excitation time (2,4,8 weeks) B2 and B3 groups, including 12 rats in group A and B, and 6 rats in groups A2, A3, B2 and B3 respectively. The proliferation of ASMCs was observed by flow cytometry, MTT assay and PCNA staining. (2) The effect of PKC activator, phorbol 12 acetate ( PMA) and inhibitor Ro 31 82 20 were used to investigate the proliferation of ASMCs in A1 and B1 groups, respectively. (3) The expression of ASMCs in groups A1, A2 and A3 were detected by reverse transcriptase-polymerase chain reaction (RT PCR) and immunocytochemistry And B1 group ASMCPKC α expression. Results (1) The ASMCS proportion, absorbance (A) value and PCNA expression in group A were significantly higher than those in group B (P <0.01). (2) The percentage of ASMCS, the value of A and the expression of PCNA in group A1 were (19 ± 3)%, 0 459 ± 0 0 36, (80 ± 10)% respectively before intervention; (14 ± 3)%, 0 346 ± 0 0 38, (53 ± 8)% and (95 ± 9)% after treatment with 50 nmol / )%; Ro 31 82 2 0 after treatment was (14 ± 3)%, 0 343 ± 0 0 48, (49 9 ± 8)%. There was significant difference between before and after treatment (P <0.01);