马旭冉, 王彦礼, 邹迪新, 刘佳星, 宋红新, 杨伟鹏, 李彧. 3种止泻名方对大鼠溃疡性结肠炎的炎性细胞因子与肠道菌群影响的对比研究J. 药学学报, 2019,54(7): 1241-1250. doi: 10.16438/j.0513-4870.2019-0051
引用本文: 马旭冉, 王彦礼, 邹迪新, 刘佳星, 宋红新, 杨伟鹏, 李彧. 3种止泻名方对大鼠溃疡性结肠炎的炎性细胞因子与肠道菌群影响的对比研究J. 药学学报, 2019,54(7): 1241-1250. doi: 10.16438/j.0513-4870.2019-0051
MA Xu-ran, WANG Yan-li, ZOU Di-xin, LIU Jia-xing, SONG Hong-xin, YANG Wei-peng, LI Yu. Comparison of intestinal bacteria and inflammatory cytokine expression in rats with ulcerative colitis after treatment of three regimentsJ. Acta Pharmaceutica Sinica, 2019,54(7): 1241-1250. doi: 10.16438/j.0513-4870.2019-0051
Citation: MA Xu-ran, WANG Yan-li, ZOU Di-xin, LIU Jia-xing, SONG Hong-xin, YANG Wei-peng, LI Yu. Comparison of intestinal bacteria and inflammatory cytokine expression in rats with ulcerative colitis after treatment of three regimentsJ. Acta Pharmaceutica Sinica, 2019,54(7): 1241-1250. doi: 10.16438/j.0513-4870.2019-0051

3种止泻名方对大鼠溃疡性结肠炎的炎性细胞因子与肠道菌群影响的对比研究

Comparison of intestinal bacteria and inflammatory cytokine expression in rats with ulcerative colitis after treatment of three regiments

  • 摘要: 对比研究黄芩汤、四神丸、痛泻要方3种止泻名方对大鼠溃疡性结肠炎(ulcerative colitis,UC)细胞因子表达与肠道菌群变化的影响。经中国中医科学院中药研究所伦理委员会审议同意并批准后,以2,4,6-三硝基苯磺酸(2,4,6-trinitrobenzenesulfonic,TNBS)复合造模法制备UC大鼠模型,按体重随机分为正常组、模型组、阳性药柳氮磺胺吡啶组(sulfasalazine,SASP,0.5g·kg-1)、黄芩汤组(HQT,20g·kg-1)、四神丸组(SSW,26g·kg-1)和痛泻要方组(TXYF,22g·kg-1)。连续给药7天后,取大鼠粪便并眼眶取血后处死,再取结肠,HE染色法检测大鼠结肠组织病变损伤程度,酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)测定血清中肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素6(interleukin-6,IL-6)、白细胞介素8(interleukin-8,IL-8)和前列腺素E2(prostaglandin E2,PGE2)含量,提取粪便样品总DNA,于16SMiseqPE300V3-4区段进行高通量测序并分析。细胞因子结果表明,与模型组相比,阳性药SASP组的IL-6、PGE2、TNF-α含量均降低(P<0.05),IL-8水平降低最为显著(P<0.01);HQT组IL-6、IL-8、TNF-α含量均降低(P<0.05),PGE2含量显著降低(P<0.01);SSW组4种炎性因子含量均降低,但无统计学差异;TXYF组IL-6、TNF-α含量降低,IL-8、PGE2含量明显降低(P<0.05)。对肠道菌群测序后的结果显示,在样本微生物信息数据量足够充分的情况下,物种丰度:SSW组 > HQT组 > TXYF组;样本间相似性:TXYF组 > SSW组 > HQT组;与模型组相比,HQT组和TXYF组菌群种类差异较大,肠内有益菌含量:HQT组 > SSW组 > TXYF组。3种名方对大鼠溃疡性结肠炎均有一定治疗作用,可有效改善UC大鼠的体征及精神状况,但在抑制炎性因子IL-6、IL-8、PGE2、TNF-α及调节肠道微生态方面,HQT的治疗作用优于SSW和TXYF。

     

    Abstract: This study was designed to compare intestinal bacteria and inflammatory cytokine expression in rats with ulcerative colitis (UC) after treatment of three regiments, Huang-qin-tang (HQT), Si-shen-wan (SSW), and Tong-xie-yao-fang (TXYF). After approved by Institute of Chinese Materia Medica Ethics Committees in China Academy of Chinese Medical Sciences, UC in rats was induced by using a compound method (trinitrobenzenesulfonic acid plus ethanol). Rats were randomly divided into control, disease, positive control salazosulfapyridine (SASP, 0.5 g·kg-1), HQT (20 g·kg-1), SSW (26 g·kg-1), and TXYF groups (22 g·kg-1). After 7 days of treatment, colonic tissues and the blood were taken for various assays. Damage of colonic tissues was detected by H&E staining. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-8 (IL-8) and prostaglandin E2 (PGE2) in the serum were detected by the enzyme linked immunosorbent assay (ELISA). Total DNA was extracted from stool samples for analyses of 16SMiseqPE300V3-4 segment using high-throughput sequencing. The inflammatory cytokine results showed that compared with the disease group, the content of IL-6, PGE2, TNF-α in SASP group were decreased (P<0.05), with the most significant decrease being the level of IL-8 (P<0.01), whereas the levels of IL-6, IL-8 and TNF-α in HQT group were reduced (P<0.05) and PGE2 content was clearly reduced (P<0.01). The contents of four cytokines in SSW group were decreased, but there was no statistical difference. While the levels of IL-6 and TNF-α in TXYF group were reduced, and the reductions of IL-8 and PGE2 were significant (P<0.05). The results after sequencing showed that microbiome species richness SSW group > HQT group > TXYF group; the similarity between samples TXYF group > SSW group > HQT group; the species of HQT and TXYF group have greater difference when compared to the disease group. The content of beneficial bacteria in the intestine of HQT group > SSW group > TXYF group. Three regiments all have therapeutic effects on UC, manifested by improvements of the signs and mental status of UC rats. However, in terms of inhibition of inflammatory factors IL-6, IL-8, PGE2 and TNF-α, and regulation of intestinal microbiome, the therapeutic effect of HQT was superior than SSW and TXYF.

     

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