平溃灌肠剂对溃疡性结肠炎肠双歧杆菌黏附性及相关炎性细胞因子的影响

Study on the bifidobacterial adhesion to the host tissue in ulcerative colitis and related serum inflammatory factors with Pingkui enema

目的观察平溃灌肠剂对溃疡性结肠炎(ulcerative colitis,UC)模型大鼠肠双歧杆菌黏附性及相关血清炎性细胞因子的影响.方法取SPF级SD大鼠60只,按照随机数字表法分为正常组,模型组,中药低、中、高剂量组,柳氮磺吡啶组.除正常组外,其余各组大鼠采用三硝基苯磺酸制备UC模型.中药低、中、高组剂量组分别灌肠平溃灌肠剂4.32、8.64、17.25 g/kg,柳氮磺吡啶组灌肠柳氮磺吡啶混悬液0.32 g/kg;正常组及模型组灌肠等体积生理盐水.1次/d,持续给药10 d.采用HE染色观察结肠组织病理学改变;采用ELISA法检测血清IL-8、IL-13、TNF-α水平及大鼠肠黏膜双歧杆菌黏附素、黏附素受体水平;采用实时荧光定量PCR法检测肠道组织双歧杆菌含量.结果与模型组比较,中药中、高剂量组IL-8[(153.50±8.59)pg/ml、(150.84±5.38)pg/ml比(167.13±13.66)pg/ml]降低,IL-13[(37.04±3.62)pg/ml、(40.64±5.98)pg/ml比(31.57±2.95)pg/ml]升高,TNF-α[(330.08±10.02)pg/ml、(287.27±13.89)pg/ml比(376.50±19.06)pg/ml]降低(P<0.05),中药低、中、高剂量组大鼠双歧杆菌黏附素[(124.84±9.74)ng/L、(149.93±9.57)ng/L、(176.74±11.01)ng/L比(101.11±17.18)ng/L]、黏附素受体[(78.44±14.03)ng/L、(99.50±3.63)ng/L、(107.36±5.05)ng/L比(60.96±13.89)ng/L]水平升高(P<0.05),中药中剂量组双歧杆菌含量[(1 331.4±242.2)拷贝数/μl比(490.5±106.0)拷贝数/μl]较模型组明显升高(P<0.05).结论平溃灌肠剂可促进双歧杆菌定植于肠上皮细胞,减轻肠黏膜屏障损伤,使促炎因子和抑炎因子恢复动态平衡,修复肠道黏膜.

Objective This study is designed to investigate the effect and mechanism of Pingkui enema in treatment of ulcerative colitis. Methods Sixty SD rats at SPF level were randomly divided into 6 groups which were normal group, model group, and the low, medium and high dose groups of Chinese medicine Pingkui enema, and sulfasalazine (SASP) enema group. The normal group and model group were given an equal volume of saline enema after modeling. After continuous administration for 10 days, the histopathological effects were evaluated after extraction, and the serum interleukin-8 (IL-8), interleukin-13 (IL-13), tumor necrosis factor (TNF-α), rat intestinal mucosal bifidobacteria adhesion and adhesin receptor were measured by enzyme-linked immunosorbent assay (ELISA). The real-time fluorescence quantitative PCR (RT-PCR) were used to detect the intestinal bifidobacterial content. Results Compared with the model group, the content of IL-8 (153.50 ± 8.59 pg/ml, 150.84 ± 5.38 pg/ml vs. 167.13 ± 13.66 pg/ml), TNF-α (330.08 ± 10.02 pg/ml, 287.27 ± 13.89 pg/ml vs. 376.50 ± 19.06 pg/ml) in medium, high dose group of Pingkui enema significantly decreased (P<0.05), while the IL-13 (37.04 ± 3.62 pg/ml, 40.64 ± 5.98 pg/ml vs. 31.57 ± 2.95 pg/ml) significantly increased (P<0.05). The bifidobacteria adhesin (124.84 ± 9.74 ng/L, 149.93 ± 9.57 ng/L, 176.74 ± 11.01 ng/L vs. 101.11 ± 17.18 ng/L) and adhesin receptor (78.44 ± 14.03 ng/L, 99.50 ± 3.63 ng/L, 107.36 ± 5.05 ng/L vs. 60.96 ± 13.89 ng/L) in low-, medium-, high dose group of Pingkui enema significantly increased (P<0.05). The content of bifidobacteria (1 331.4 ± 242.2 copies/μl vs. 490.5 ± 106.0 copies/μl) in the middle dose Pingkui enema group significantly increased (P<0.05). Conclusions The Pingkui enema can promote the colonization of intestinal epithelial cells by promoting bifidobacteria, increase bifidobacterium adhesin and adhesin receptors, up-regulate the expression of IL-13, and down-regulate the expression of IL-8, TNF-α.