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目的观察在内毒素休克及白介素-1β介导家兔肠系膜上动脉(superior mesenteric artery,SMA)钙敏感性改变中SMA PKC、Rho激酶活性的变化。方法健康成年家兔96只,雌雄不拘,分为两实验大组:①内毒素休克组(包括5小组:正常对照组、LPS 30 min组、LPS 1 h组、LPS 2 h组和LPS 4 h组);②IL-1β处理组(包括3小组:0 ng/ml IL-1β组、50 ng/ml IL-1β组、100 ng/ml IL-1β组),每小组12只家兔。经耳缘静脉注入1 mg/kg LPS(O111∶B4)复制内毒素休克模型,在给LPS前及给LPS后30 min,1、2、4 h取家兔SMA,提取其PKC及Rho激酶蛋白并纯化,检测PKC及Rho激酶活性的变化,并与内毒素休克后血管钙敏感性变化进行相关分析;无菌条件下取正常成年家兔SMA,在0、50、100 ng/ml IL-1β作用12 h后,检测其PKC及Rho激酶活性的变化,并与IL-1β处理后家兔SMA钙敏感性变化进行相关分析。结果在给LPS后30 min至1 h成功复制出内毒素休克模型,与正常对照组相比,PKC、Rho激酶活性均在给LPS 30 min后升高(P<0.01),在2 h及4 h后均降低(P<0.01),与钙敏感性变化均呈显著正相关(r=0.912和0.968,P<0.05)。与0 ng/mlIL-1β组相比,100 ng/ml IL-1β能使PKC、Rho激酶活性明显降低(P<0.05),与钙敏感性变化均呈显著正相关(r=0.922和0.934,P<0.05)。结论 IL-1β能通过改变PKC、Rho激酶活性来调节家兔肠系膜上动脉钙敏感性,IL-1β可能通过改变PKC、Rho激酶活性调节家兔内毒素休克后肠系膜上动脉钙敏感性变化。
Objective To observe the changes of SMA PKC and Rho kinase in endotoxin shock and interleukin-1β-mediated calcium sensitivity change in rabbit superior mesenteric artery (SMA). Methods 96 healthy adult rabbits were randomly divided into two groups: ① endotoxin shock group (including 5 groups: normal control group, LPS 30 min group, LPS 1 h group, LPS 2 h group and LPS 4 h Group); ② IL-1β treatment group (including 3 groups: 0 ng / ml IL-1β group, 50 ng / ml IL-1β group, 100 ng / ml IL-1β group), each group of 12 rabbits. The model of endotoxic shock was established by injecting 1 mg / kg LPS (O111: B4) into the ear vein. Rabbit SMA was harvested before LPS administration and at 30 min, 1, 2 and 4 h after LPS injection. PKC and Rho kinase protein The changes of PKC and Rho kinase activity were detected by enzyme linked immunosorbent assay (ELISA). The changes of calcium sensitivity in vascular smooth muscle cells after endotoxic shock were also analyzed. Normal adult rabbit SMA was treated with 0, 50 and 100 ng / ml IL-1β After 12 h, the changes of PKC and Rho kinase activity were detected, and the changes of SMA calcium sensitivity in rabbit after IL-1β treatment were analyzed. Results Endotoxic shock models were successfully replicated 30 min to 1 h after LPS administration. PKC and Rho kinase activity increased at 30 min after LPS administration (P <0.01). Compared with normal control group, h (P <0.01), and there was a significant positive correlation between calcium sensitivity and calcium sensitivity (r = 0.912 and 0.968, P <0.05). Compared with 0 ng / ml IL-1β group, 100 ng / ml IL-1β significantly decreased the PKC and Rho kinase activity (P <0.05), and had a positive correlation with calcium sensitivity (r = 0.922 and 0.934, P <0.05). Conclusion IL-1β can regulate the calcium sensitivity of rabbit superior mesenteric artery by changing PKC and Rho kinase activity. IL-1β may regulate the calcium sensitivity of superior mesenteric artery after endotoxin shock by changing PKC and Rho kinase activity.