清胰II号对重症急性胰腺炎大鼠肠道免疫损伤的影响
| 作者: |
1周淼,
2代静静,
1兑丹华,
1敖宇
1 遵义医学院附属医院 普通外科，贵州 遵义563000 2 遵义市第一人民医院 重症监护室，贵州 遵义 563000 |
| 通讯: |
周淼
Email: 1234567@qq.com 代静静 Email: 123@qq.com 兑丹华 Email: 2361169015@qq.com 敖宇 Email: 123@qq.com |
| DOI: | 10.3978/.10.3978/j.issn.1005-6947.2017.03.008 |
| 基金: | 贵州省科学技术基金资助项目, 黔科合J字2008-2200号 贵州省遵义市红花岗区科技局基金资助项目, fcz120131222 |
摘要
目的：探讨清胰II号对重症急性胰腺炎（SAP）大鼠肠道免疫损伤的影响。方法：将80只SD大鼠随机分成假手术组、SAP模型组（SAP组）、SAP模型+清胰II号治疗组（清胰II号组）、SAP模型+阳性药物对照组（谷氨酰胺组），其中假手术组8只，其余组各24只；采用胰胆管逆行注射5%的牛磺胆酸钠建立SAP模型；清胰II号组与谷氨酰胺组术后分别用清胰II号（10 mL/kg，1次/6 h）和谷氨酰胺灌胃（0.15 g/100 g，1次/6 h）灌胃，另两组以相同的方式给予等体积的生理盐水；除假手术组大鼠在术后6 h采集标本外，其余各组均分别在术后6、12、24 h各取8只大鼠采集标本，观察胰腺与回肠组织的病理变化；ELISA法检测血清IL-1、IL-10浓度；RT-PCR检测回肠组织HMGB1 mRNA的表达；流式细胞仪检测回肠Peyer结中CD3+、CD4+、CD8+T淋巴细胞亚群的凋亡。结果：除假手术组外，其余各组胰腺与回肠组织均出现明显的病理学改变，且逐渐加重，但两个治疗组术后各时间点胰腺与回肠组织损伤程度均轻于SAP组。与对照组比较，其余各组术后血清IL-1、IL-10水平、回肠HMGB1 mRNA表达量均明显逐渐升高（均P<0.05），但两个治疗组IL-1、IL-10的升高幅度均低于SAP组（均P<0.05）；其余各组术后回肠组织CD3+、CD4+、CD8+T淋巴细胞凋亡率均明显升高（均P<0.05），其中在SAP组呈逐渐升高趋势，而在两个治疗组呈逐渐降低趋势，且两个治疗组各时间点的各T细胞的凋亡率均小于SAP组（均P<0.05）。两个治疗组间同时间点上述各指标的差异均无统计学意义（均P>0.05）。结论：清胰II号可减轻SAP时肠道免疫功能的损伤，其机制可能与下调回肠HMGB1表达，减少T淋巴细胞的凋亡有关。
关键词：
胰腺炎，急性坏死性
肠黏膜
炎症
细胞凋亡
方剂
Effect of Qingyi II formula on intestinal immune injury secondary to severe acute pancreatitis in rats
CorrespondingAuthor:ZHOU Miao Email: 1234567@qq.com
Abstract
Objective: To investigate the effect of Qingyi II formula on intestinal immune injury secondary to severe acute pancreatitis in rats. Methods: Fifty-six SD rats were randomly divided into sham operation group, SAP model group (SAP group), SAP model plus Qingyi II formula treatment group (Qingyi II group), and SAP model plus positive drug control group (glutamine group), with 8 rats in sham operation group and 24 rats each in the other groups. SAP model was induced by retrograde injected 5% sodium taurocholate into the bile-pancreatic duct. After operation, rats in Qingyi II group and glutamine group were subjected to gavage administration of Qingyi II formula (10 mL/kg,
once per 6 h) and glutamine (0.15 g/100 g, once per 6 h) respectively, while those in the other two groups were given the same volume of normal saline instead in the same fashion. The specimen samples, except in sham operation group that were harvested at 6 h after operation, were harvested at 6, 12 and 24 h after operation in all of the other groups, with 8 rats in each time point. Then, the pathological changes in pancreatic and ileal tissues were observed, the serum concentrations of IL-1 and IL-10 were determined by ELISA assay, the HMGB1 mRNA expressions in the ileal tissues were detected by RT-PCR, and the apoptosis of CD3+, CD4+ and CD8+T cell subsets in the Peyer’s patches of the ileum were measured by flow cytometry. Results: Except in sham operation group, obvious and gradually aggravated pathological changes were seen in the pancreatic and ileal tissues in all of the other groups, which were all milder in both treatment groups than those in SAP group at each postoperative time point. Compared with sham operation group, the serum concentrations of IL-1 and IL-10, and HMGB1 mRNA expressions in the ileal tissues were all significantly and increasingly elevated in the remaining groups (all P<0.05), but the increasing amplitudes of IL-1 and IL-10 in both treatment groups were milder than those in SAP group at each postoperative time point (all P<0.05); the apoptosis rates of CD3+, CD4+ and CD8+T lymphocytes in the ileal tissues were all significantly increased in the remaining groups (all P<0.05), which showed an ascending trend in SAP group while a descending trend in either treatment group,and further, the apoptosis rates of these T lymphocytes in either treatment group were significantly lower than those in SAP group at each time point (all P<0.05). The difference in all above parameters showed no statistical significance between the two treatment group at the same time points (all P>0.05). Conclusion: Qingyi II formula can lessen the intestinal immune injury secondary to SAP in rats, and the mechanism may be associated with its down-regulating of HMGB1 expression in the ileum and thereby decreasing the apoptosis of T lymphocytes.
Keywords：
once per 6 h) and glutamine (0.15 g/100 g, once per 6 h) respectively, while those in the other two groups were given the same volume of normal saline instead in the same fashion. The specimen samples, except in sham operation group that were harvested at 6 h after operation, were harvested at 6, 12 and 24 h after operation in all of the other groups, with 8 rats in each time point. Then, the pathological changes in pancreatic and ileal tissues were observed, the serum concentrations of IL-1 and IL-10 were determined by ELISA assay, the HMGB1 mRNA expressions in the ileal tissues were detected by RT-PCR, and the apoptosis of CD3+, CD4+ and CD8+T cell subsets in the Peyer’s patches of the ileum were measured by flow cytometry. Results: Except in sham operation group, obvious and gradually aggravated pathological changes were seen in the pancreatic and ileal tissues in all of the other groups, which were all milder in both treatment groups than those in SAP group at each postoperative time point. Compared with sham operation group, the serum concentrations of IL-1 and IL-10, and HMGB1 mRNA expressions in the ileal tissues were all significantly and increasingly elevated in the remaining groups (all P<0.05), but the increasing amplitudes of IL-1 and IL-10 in both treatment groups were milder than those in SAP group at each postoperative time point (all P<0.05); the apoptosis rates of CD3+, CD4+ and CD8+T lymphocytes in the ileal tissues were all significantly increased in the remaining groups (all P<0.05), which showed an ascending trend in SAP group while a descending trend in either treatment group,and further, the apoptosis rates of these T lymphocytes in either treatment group were significantly lower than those in SAP group at each time point (all P<0.05). The difference in all above parameters showed no statistical significance between the two treatment group at the same time points (all P>0.05). Conclusion: Qingyi II formula can lessen the intestinal immune injury secondary to SAP in rats, and the mechanism may be associated with its down-regulating of HMGB1 expression in the ileum and thereby decreasing the apoptosis of T lymphocytes.