目的 研究黄连-厚朴配伍对溃疡性结肠炎（ulcerative colitis，UC）大鼠及磷脂酰肌醇-3-激酶（phosphatidylinositol-3-kinase，PI3K）/蛋白激酶B（protein kinase B，Akt）信号通路的调控作用。方法 利用高通量基因表达数据库获取UC基因表达谱数据，并综合生物信息学分析，预测黄连-厚朴治疗UC的潜在机制。采用2，4，6-三硝基苯磺酸（TNBS）/乙醇法建立UC大鼠模型，SD大鼠随机分为对照组、模型组、黄连（2 g/kg）组、厚朴（2 g/kg）组、黄连-厚朴配伍（4 g/kg）组和柳氮磺胺吡啶（0.4 g/kg）组，每组8只。造模24 h后，各给药组ig相应药物，1次/d，连续6 d，采用试剂盒检测各组大鼠血清中白细胞介素-1β（interleukin-1β，IL-1β）、IL-6、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）水平和髓过氧化物酶（myeloperoxidase，MPO）活性；计算并评价各组大鼠结肠质量/长度、脾脏指数和肠黏膜损伤指数（colon macroscopic damage index，CMDI）评分；采用苏木素-伊红（HE）染色法观察各组大鼠结肠组织病理变化并进行病理损伤评分；采用免疫组化法检测各组大鼠结肠组织PI3K、Akt和剪切型半胱氨酸蛋白酶-3（cleaved Caspase-3）表达情况；采用Western blotting法检测各组大鼠结肠组织磷酸化Akt（p-Akt）、B淋巴细胞瘤2（B-cell lymphoma 2，Bcl-2）和cleaved Caspase-3蛋白表达情况。结果 生物信息学预测显示，黄连-厚朴抗UC的潜在靶点有49个，可能与调控生物过程及过氧化物酶体增殖物活化受体（peroxisome proliferator-activated receptor，PPAR）信号通路、核因子κB（nuclear factor-κB，NF-κB）信号通路和PI3K/Akt信号通路等有关。实验验证结果显示，与模型组比较，各给药组大鼠血清中IL-1β、IL-6、TNF-α水平和MPO活性显著降低（P<0.01）；结肠质量/长度、脾脏指数、CMDI评分、结肠组织病理损伤评分均显著降低（P<0.01）；结肠组织PI3K、Akt和cleaved Caspase-3阳性表达显著减少（P<0.01）；结肠组织中p-Akt/Akt、cleaved Caspase-3蛋白表达水平显著降低（P<0.01），Bcl-2蛋白表达水平显著升高（P<0.01），其中黄连-厚朴配伍组疗效最佳。结论 黄连、厚朴及其配伍能够有效缓解TNBS诱导的UC，其作用机制可能与抑制PI3K/Akt通路相关，且黄连-厚朴配伍疗效最佳，体现了中药“相使”增效的配伍关系。

Objective To study the effect of Huanglian (Coptidis Rhizoma)-Houpo (Magnoliae Officinalis Cortex) on regulation of phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in rats with ulcerative colitis (UC). Methods The potential mechanism of Coptidis Rhizoma-Magnoliae Officinalis Cortex on treating UC was predicted based on UC gene expression profile data obtained from high-throughput gene expression database combined with comprehensive bioinformatics analysis. UC rats model was established by 2,4,6-trinitrobenzene sulfonic acid (TNBS)/ethanol method. SD rats were randomly divided into control group, model group, Coptidis Rhizoma (2 g/kg) group, Magnoliae Officinalis Cortex (2 g/kg) group, Coptidis Rhizoma-Magnoliae Officinalis Cortex (4 g/kg) group and sulfasalazine (0.4 g/kg) group, with eight rats in each group. The rats in each administration group were ig corresponding drugs 24 h after modeling, once a day for 6 d. Levels of interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α) and myeloperoxidase (MPO) activity in serum of rats in each group were detected by kits; Colon mass/length, spleen index and colon macroscopic damage index (CMDI) were calculated and evaluated; Hematoxylin-eosin (HE) staining method was used to investigate the pathological changes and pathological damage scores of colon tissue in rats; Immunohistochemical method was used to detect PI3K, Akt and cleaved Caspase-3 expression in colon of rats; Western blotting was used to detect expressions of phosphorylated Akt (p-Akt), B-cell lymphoma 2 (Bcl-2) and cleaved Caspase-3 in colon of rats. Results Bioinformatics predictions showed that there were 49 potential targets of Coptidis Rhizoma-Magnoliae Officinalis Cortex on UC, which may be related to the regulation of biological processes and pathways such as peroxisome proliferator-activated receptor (PPAR) signaling pathway, nuclear factor-κB (NF-κB) signaling pathway and PI3K/Akt signaling pathway. Experimental validation results showed that compared with model group, IL-1β, IL-6, TNF-α levels and MPO activity in serum of rats in each administration group were significantly reduced (P < 0.01); Colon mass/length, spleen index, CMDI score, and colon tissue pathological damage score of rats were significantly reduced (P < 0.01); Positive expressions of PI3K, Akt and cleaved Caspase-3 in colon tissue was significantly reduced (P < 0.01); Expressions of p-Akt/Akt and cleaved Caspase-3 in colon were significantly reduced (P < 0.01), Bcl-2 expression was significantly increased (P < 0.01), and Coptidis Rhizoma-Magnoliae Officinalis Cortex had the best curative effect. Conclusion Coptidis Rhizoma, Magnoliae Officinalis Cortex and their compatibility can effectively alleviate TNBS-induced UC, of which mechanism may be related to the inhibition of PI3K/Akt pathway, and Coptidis Rhizoma-Magnoliae Officinalis Cortex has the best effect, which reflects the compatibility of traditional Chinese medicines with “combination”.

R285.5

成都中医药大学中药“性-效-用”理论与实践创新团队（CXTD2018004）；成都中医药大学西南特色中药资源重点实验室开放研究基金资助项目（2020XSGG025）