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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(10mL/kg,1次/6h)和谷氨酰胺灌胃(0.15g/100g,1次/6h)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6h采集标本外,其余各组均分别在术后6、12、24h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HMGB1m RNA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。
Objective: To investigate the effect of Qingyi No.2 on intestinal immune injury in rats with severe acute pancreatitis (SAP). Methods: Eighty SD rats were randomly divided into sham operation group, SAP model group (SAP group), SAP model + Qingyiyi Ⅱ treatment group (Qingyi Ⅱ group), SAP model + positive drug control group (glutamine Group), including sham operation group (n = 8) and the remaining group (n = 24). The SAP model was established by retrograde injection of pancreaticobiliary duct with 5% sodium taurocholate. Qingjian II group and glutamine group (10mL / kg, 1 / 6h) and glutamine gavage (0.15g / 100g, 1 / 6h), the other two groups in the same way to give an equal volume of saline; in addition to the sham group The rats were collected at 6h after operation. All the other groups were collected at 6, 12, and 24 hours after operation, respectively, and the pathological changes of pancreas and ileum were observed. The levels of IL-1, IL- 10 concentration. The expression of HMGB1 mRNA in ileum was detected by RT-PCR. The apoptosis of CD3 +, CD4 +, CD8 + T lymphocyte subsets in Peyer’s node of ileum was detected by flow cytometry. Results: In addition to the sham operation group, the pathological changes of pancreas and ileum in other groups were obvious and gradually increased. However, the damage degree of pancreas and ileum in the two treatment groups was lighter than that in SAP group. Compared with the control group, the levels of IL-1, IL-10 and HMGB1 mRNA in the other groups were significantly increased (all P <0.05), but the levels of IL-1, IL-10 (P <0.05). The apoptosis rates of CD3 +, CD4 +, CD8 + T lymphocytes in the other groups were significantly increased (all P <0.05) Which increased gradually in SAP group, but gradually decreased in both groups. The apoptotic rates of T cells in both groups were lower than those in SAP group at each time point (P <0.05). There was no significant difference between the two treatment groups at the same time point (P> 0.05). Conclusion: QingYi Ⅱ can relieve the intestinal immune function injury in SAP. The mechanism may be related to down-regulation of HMGB1 expression in the ileum and reduction of T lymphocyte apoptosis.