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目的探讨蛋白酶激活受体-2(proteinase-actived receptors-2,PAR-2)mRNA和蛋白在小鼠胃肠道各部位的表达情况及PAR-2激动剂对小鼠胃肠动力的影响。方法 130只BABL/c小鼠,禁食不禁水16~24h后,取10只小鼠胃、十二指肠、空肠及回肠组织,采用RT-PCR法检测各部位PAR-2 mRNA表达情况,采用免疫组织化学法检测各部位PAR-2蛋白表达情况。余120只小鼠随机分为6组,分别为对照组、抑氨肽酶素A对照组(抑氨肽酶素A组)、SLIGRL-NH_2 1μmol/kg+抑氨肽酶素A组(SLIGRL-NH_2 1μmol/kg组)、SLIGRL-NH_2 2.5μmol/kg+抑氨肽酶素A组(SLIGRL-NH_2 2.5μmol/kg组)、SLIGRL-NH_2 5μmol/kg+抑氨肽酶素A组(SLIGRL-NH_2 5μmol/kg组)、LRGILS-NH_2 5μmol/kg+抑氨肽酶素A组(LRGILS-NH_2 5μmol/kg组),每组20只。各组分别腹腔注射实验分组的相应药物,随后迅速给予葡聚糖蓝2000 0.4 mL灌胃,测定并比较6组小鼠小肠推进比和胃内色素排空率。结果PAR-2在小鼠胃、十二指肠、空肠及回肠均有表达;SLIGRL-NH_22.5μmol/kg组和SLIGRL-NH_2 5μmol/kg组小鼠胃内色素排空率[(73.65±8.61)%、(74.81±8.58)%]和小肠推进比[(62.75±3.52)%、(66.38±4.92)%]明显高于对照组[(62.13±7.82)%、(42.32±4.02)%]、抑氨肽酶素A组[(61.72±8.32)%、(43.21±5.12)%]和SLIGRL-NH_2 1μmol/kg组[(61.92±8.96)、(41.73±3.98)%](P<0.05),SLIGRL-NH_2 2.5μmol/kg组与SLIGRL-NH_25μmol/kg组比较差异无统计学意义(P>0.05);对照组、抑氨肽酶素A组、SLIGRL-NH_2 1μmol/kg组和LRGILS-NH_2 5μmol/kg组小肠推进比、胃内色素排空率两两比较差异均无统计学意义(P>0.05)。结论 PAR-2在小鼠胃肠道中广泛表达,在调节消化道功能中起重要作用,激活后可明显提高小鼠的胃肠动力。
Objective To investigate the expression of PAR-2 mRNA and protein in various parts of the gastrointestinal tract in mice and the effect of PAR-2 agonist on gastrointestinal motility in mice. Methods Totally 130 BABL / c mice were fasted for 16-24 h after fasting. The gastric mucosa, duodenum, jejunum and ileum of 10 mice were sacrificed. The expression of PAR-2 mRNA was detected by RT-PCR. Immunohistochemistry was used to detect the expression of PAR-2 protein. The remaining 120 mice were randomly divided into 6 groups: control group, aminopeptidase A control group (aminopeptidase A group), SLIGRL-NH 2 1 μmol / kg + aprotinin group A (SLIGRL- (SLIGRL-NH 2 2.5μmol / kg), SLIGRL-NH 2 5μmol / kg + aminoglitazone A (SLIGRL-NH 2 5μmol / kg), SLIGRL-NH 2 2.5μmol / kg + / kg group), LRGILS-NH 2 5μmol / kg + aminoglitazone group A (LRGILS-NH 2 5μmol / kg group), with 20 rats in each group. Each group was intraperitoneal injection of the corresponding group of drugs, followed by rapid dextran blue 2000 0.4 mL gavage, determination and comparison of 6 groups of mice intestinal propulsion ratio and the rate of gastric emptying. Results PAR-2 was expressed in the stomach, duodenum, jejunum and ileum of mice. The gastric emptying rate in SLIGRL-NH_22.5μmol / kg group and SLIGRL-NH_2 5μmol / kg group [(73.65 ± 8.61 ), (74.81 ± 8.58)% and small intestine propulsion rate (62.75 ± 3.52)% and (66.38 ± 4.92)%, respectively, were significantly higher than those in the control group (62.13 ± 7.82% and 42.32 ± 4.02%, respectively) (61.72 ± 8.32)%, (43.21 ± 5.12)%] and SLIGRL-NH_2 1μmol / kg group [(61.92 ± 8.96), (41.73 ± 3.98)%] (P <0.05) There was no significant difference between SLIGRL-NH_2 2.5μmol / kg group and SLIGRL-NH_25μmol / kg group (P> 0.05). In the control group, the group treated with amitriptyline A, SLIGRL-NH_2 1μmol / kg and LRGILS-NH 2 5μmol / kg group of intestinal propulsion ratio, gastric emptying rate of any significant difference between the two groups were not statistically significant (P> 0.05). Conclusion PAR-2 is widely expressed in the gastrointestinal tract of mice and plays an important role in the regulation of gastrointestinal function. Activation of PAR-2 can significantly improve the gastrointestinal motility of mice.