新型大麻制剂O-1602对急性胰腺炎小鼠肠道免疫及菌群的影响

目的 研究新型大麻制剂O-1602对急性胰腺炎(acute pancreatitis, AP)小鼠肠道免疫及菌群紊乱的干预作用。方法 成年C57/BL6小鼠21只,雌雄各半,随机均分为AP模型组(AP组,n=7)、AP治疗组(AP+O-1602组,n=7)及正常对照组(control组,n=7)。AP模型组、AP治疗组以雨蛙肽腹腔注射(50μg/kg)复制AP模型,并分别给以腹腔注射药物溶剂或O-1602溶液(10mg/kg)；正常对照组小鼠用相同方式获得等量生理盐水及药物溶剂注射。3组动物均于末次注药3h后麻醉处死,收集其血液、小肠和结肠及肠道Peyer's结。检测和分析肠道细菌总数和主要类别、Peyer's结中T淋巴细胞及其亚群的变化；检测血浆淀粉酶及细胞因子IL-10、MCP-1水平的变化。结果 与对照组相比,AP小鼠肠道细菌总数、主要亚群,尤其是大肠杆菌、肠球菌的数量、血浆淀粉酶活性、IL-10及MCP-1水平都有明显升高(P＜0.05或P＜0.01)；新型大麻制剂O-1602可在一定程度上逆转AP时的上述异常变化(P＜0.05)。各组肠道Peyer's结数量无明显差异,但AP小鼠Peyer's结中CD3+、CD4+T淋巴细胞百分比较对照组明显降低(AP组P＜0.01,AP+O-1602组P＜0.05),CD4+/CD8+较对照组也有所降低,但差异无统计学意义。结论 O-1602对雨蛙肽诱导的小鼠急性胰腺炎具有一定拮抗作用,其机制可能与其促进肠道细菌的清除、调节肠道T淋巴细胞功能、抑制炎症反应等有关。
Objective To investigate effect of a novel cannabis preparation O-1602 on the intestinal immunity and microflora in mice with acute pancreatitis(AP). Methods Twenty one adult C57/BL6 mice were randomly divided into three group: normal control group(control, n=7), AP group(n=7), and O-1602 treated group(AP+O-1602, n=7). AP was induced by intraperitoneal injection of caerulein(50μg /kg). Drug vehicle or O-1602(10mg/kg) was given intraperitoneal injection to AP group or AP+O-1602 group respectively, and equivalent volume of normal saline and drug vehicle was given to the control group. The mice were sacrificed at 3h after the last injection, and the blood, the whole small intestine and colon, the intestinal Peyer's patches were harvested. The microflora in the ileum and colon were examined； the T lymphocytes and its subgroups in the Peyer's patches were determined. The plasma levels of amylase, interleukin-10(IL-10) and monocyte chemotactic protein 1(MCP-1) were measured. Results Compared to the control group, mice in AP group presented with significantly increased intestinal microflora, plasma amylase activity and levels of IL-10 and MCP-1(P＜0.05 or P＜0.01). O-1602 attenuated these changes to some extent(P＜0.05). There was no significant difference in the numbers of Peyer's patches in the intestine among the three groups； but the percentage of CD3+ , CD4+ T lymphocyte decreased significantly in AP group and AP+O-1602 group(P＜0.01 and P＜0.05, respectively), and an indistinctively decreased percentage of CD4+/ CD8+ was observed. Conclusion O-1602 protects mice from cearulein-induced acute pancreatitis, which may be related to suppressing inflammatory response, promoting bacteria removal, and regulating T lymphocyte function in intestinal barrier