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目的探讨泻剂结肠大鼠结肠中阿片受体信号调节相关蛋白RGS-4和β-arrestin2的表达变化及意义。方法 7~8周龄Wistar大鼠16只,体质量(200±20)g,雌雄各半,按随机数字表法分为对照组(n=8)及泻剂结肠组(n=8),饲养环境温度18~28℃,相对湿度40%~80%。对照组饲以普通软饲料,泻剂结肠组饲以混有酚酞的饲料,建立泻剂结肠大鼠模型。通过RT-PCR及Western blot方法对RGS-4和β-arrestin2在结肠中的表达进行相对定量分析。结果两种阿片受体信号调节蛋白在泻剂结肠组及对照组大鼠结肠内均有不同程度的表达。RT-PCR结果显示泻剂结肠组RGS-4和β-arrestin2 mRNA相对表达水平均明显高于对照组[(3.418 3±0.247 4)vs(0.987 6±0.034 1),(2.974 4±0.214 2)vs(0.921 1±0.040 1),P<0.01]。Western blot检测结果显示RGS-4和β-arrestin2的表达在泻剂结肠组较对照组明显升高[(0.403 4±0.049 9)vs(0.115 3±0.010 2),(0.913 9±0.061 1)vs(0.467 6±0.043 7),P<0.01]。结论RGS-4和β-arrestin2在泻剂结肠大鼠结肠中表达均明显增强,提示其可能增强泻剂结肠大鼠结肠MOR信号传导调节功能。
Objective To investigate the expression and significance of opioid receptor signaling-related proteins RGS-4 and β-arrestin2 in colon of laxative colon rats. Methods Sixteen Wistar rats, aged 7-8 weeks old, were divided into control group (n = 8) and cathartic colon group (n = 8) according to the random number table. Rearing environment temperature 18 ~ 28 ℃, relative humidity 40% ~ 80%. The control group was fed ordinary soft feed, cathartic colon group fed with phenolphthalein feed, laxative colon rat model was established. The relative quantitative expression of RGS-4 and β-arrestin2 in the colon was analyzed by RT-PCR and Western blot. Results Both opioid receptor signaling proteins were expressed in the colon of the cathartic colon and the control rats. RT-PCR results showed that the relative expression levels of RGS-4 and beta-arrestin2 mRNA in the cathartic colon group were significantly higher than those in the control group [(3.418 3 ± 0.247 4) vs (0.987 6 ± 0.034 1), (2.974 4 ± 0.214 2) vs (0.921 1 ± 0.040 1), P <0.01]. The results of Western blot showed that the expressions of RGS-4 and β-arrestin2 were significantly increased in the cathartic colon as compared with the control group [(0.403 4 ± 0.049 9) vs (0.115 3 ± 0.010 2), (0.913 9 ± 0.061 1) vs (0.467 6 ± 0.043 7), P <0.01]. Conclusion The expression of RGS-4 and β-arrestin2 in the colon of laxative colon rats were significantly increased, suggesting that it may enhance colon MOR signaling regulation.