中文摘要：

目的探索在内脏高敏感状态下,跨器官敏化模型鼠脊髓背角蛋白激酶（PKC）γ基因表达的改变并探索其与内脏痛敏程度之间的相关性,寻求调控跨器官敏化的新靶点。方法采用鸡卵清蛋白（OVA）基础致敏联合结直肠芥子油（MO）灌注的方法建立跨器官敏化大鼠模型。30只SD大鼠随机分为：正常对照组（A组,n=7）;跨器官敏化模型组（B组,n=8）,给予OVA基础致敏联合MO结肠灌注;OVA＋MO＋生理盐水对照组（C组,n=7）,给予OVA基础致敏联合MO结肠灌注,鞘内注射生理盐水10μl;OVA＋MO＋PKCγ特异性抑制剂（GF109203X）组（D组,n=8）,OVA基础致敏联合MO结肠灌注,鞘内注射GF109203X10μl。采用免疫荧光染色技术及Westernblotting检测各组大鼠脊髓PKCγ的表达情况,并探讨PKCγ表达与内脏痛敏程度之间的关系。结果免疫荧光染色结果显示,跨器官敏化模型组大鼠L4-6脊髓背角组织PKCγ蛋白表达量（98.99±7.73）明显多于正常对照组（56.14±6.22,P〈0.05）;而鞘内注射GF109203X的D组大鼠PKCγ表达量显著降低,明显低于鞘内注射生理盐水的C组（56.83±7.07vs89.59±9.90,P〈0.05）。Westernblotting检测结果也显示,B组大鼠L4-6脊髓背角组织中PKCγ的表达量（1.538±0.228）与A组（1.065±0.064）相比显著增加（P〈0.001）;D组大鼠PKCγ表达量（1.124±0.169）与C组大鼠（1.605±0.224）比较则显著降低（P〈0.001）。结论采用OVA基础致敏联合MO结肠灌注可成功建立跨器官敏化大鼠模型,脊髓背角组织中PKCγ参与了跨器官内脏痛敏信号的传递,在跨器官敏化的产生和维持中发挥了重要作用。

英文摘要：

Objective To investigate the changes in PKC gamma（PKCγ）expression in spinal cord dorsal horn in the cross-organ sensitized rat model,and study the relationship between PKCγexpression and visceral hypersensitivity.Methods Ovalbumin（OVA） basal-sensitization combined with colon mustard oil（MO）instillation was used to reproduce the sensitized cross-organ rat model.Thirty SD rats were randomly divided into four groups：control group（group A,n=7）,cross-organ sensitized model group（group B,received basal OVA-sensitization combined with colon MO instillation,n=8）,OVA＋MO＋saline group（group C,received basal OVA-sensitization combined with colon MO instillation and intrathecal injection of 10lsaline,n=7）,OVA＋MO＋GF109203Xgroup（group D,received basal OVA-sensitization combined with colon MO instillation and intrathecal injection of 10lGF109203X,n=8）.The expression of PKCγin spinal cord dorsal horn was monitored by Western blotting and immunofluorescence assay.The relationship between PKCγexpression and visceral hypersensitivity was analyzed.Results It was shown by immunofluorescence assay that the PKCγexpression in L4-6of spinal cord was significantly higher in cross-organ sensitized model rats（98.99±7.73）than in control rats（56.14±6.22,P〈0.05）.The expression of PKCγ in group D（56.83±7.07）was decreased significantly compared with that in group C（89.59±9.90,P〈0.05）.Western blotting analysis also showed that the PKCγexpression was significantly increased in group B（1.538±0.228）compared with that in group A（1.065±0.064, P〈0.001）,while significantly lower in group D compared with group C（1.124±0.169 vs 1.605±0.224）.Conclusions The cross-organ sensitized rat model can be successfully established by basic OVA-sensitization combined with colon MO instillation.PKCγplays an important role in the development and maintenance of colon-urethra cross-organ sensitization by involving in the transmission of hyperalgesia signal.