左金丸对DSS诱导的溃疡性结肠炎的作用及其机制

Effect and Mechanism of Zuojinwan on DSS-induced Ulcerative Colitis

目的:基于网络药理学与实验验证探究左金丸(ZJW)治疗溃疡性结肠炎(UC)的作用及机制。方法:借助网络药理学与分子对接初步明确ZJW治疗UC的活性成分与潜在机制,将48只雄性C57BL/6J小鼠随机分为正常组、模型组、柳氮磺吡啶组(300 mg·kg^(-1))和左金丸低、中、高剂量组(1.82、3.64、7.28 g·kg^(-1)),采用葡聚糖硫酸钠(DSS)构建UC模型,造模第3天开始灌胃给药,连续7 d,每日观察小鼠一般状态并评估疾病活动指数(DAI)。苏木素-伊红(HE)染色观察结肠组织病理学变化,酶联免疫吸附测定法(ELISA)检测小鼠血清中肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6和IL-10水平,并以蛋白免疫印迹法(Western blot)验证分子机制。结果:网络药理学预测发现磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路可能是ZJW调节UC的关键途径。分子对接结果表明ZJW关键成分与核心靶点之间均有良好的结合能力。动物研究结果显示,与正常组比较,模型组小鼠结肠长度显著缩短(P<0.01),DAI评分、脾脏指数、结肠组织病理学评分与血清中TNF-α、IL-6水平明显升高(P<0.05,P<0.01),结肠组织PI3K、磷酸化蛋白激酶B(p-Akt)与B细胞淋巴瘤-2(Bcl-2)相关X蛋白(Bax)表达明显增加(P<0.05,P<0.01),血清IL-10水平与结肠组织Bcl-2蛋白表达显著降低(P<0.01);与模型组比较,左金丸各给药组小鼠UC症状明显改善,结肠组织病理损伤减轻,血清中炎性细胞因子TNF-α、IL-6水平显著下调(P<0.01),结肠组织PI3K、p-Akt与Bax蛋白表达明显抑制(P<0.05,P<0.01),血清IL-10水平与结肠组织Bcl-2蛋白表达显著提高(P<0.01),以高剂量组效果最佳。结论:ZJW可有效缓解DSS诱导的UC,其机制可能与抑制PI3K/Akt信号通路、调节凋亡相关蛋白表达相关。

Objective:To explore the effect and mechanism of Zuojinwan(ZJW)in the treatment of ulcerative colitis(UC)through network pharmacology and experimental validation.Method:Using network pharmacology and molecular docking,the active components and potential mechanism of ZJW in treating UC were preliminarily identified.Forty-eight male C57BL/6J mice were randomly divided into a normal group,a model group,a sulfasalazine group(300 mg·kg^(-1)),and low-,medium-,and high-dose ZJW groups(1.82,3.64,7.28 g·kg^(-1)).The UC model was induced by dextran sulfate sodium(DSS),and oral administration of drugs began on the third day of modeling,lasting for 7 days.The general condition of mice was observed daily,and the disease activity index(DAI)was evaluated.Hematoxylin-eosin(HE)staining was performed to observe histopathological changes in colon tissue.Enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of tumor necrosis factor(TNF)-α,interleukin(IL)-6,and IL-10 in mouse serum.The molecular mechanism was validated using Western blot.Result:Network pharmacology predicted that the phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)signaling pathway might be a key pathway in the regulation of UC by ZJW.Molecular docking results showed good binding ability between the key components of ZJW and core targets.Animal experiment results showed that compared with the normal group,the model group had shortened colon length(P<0.01),increased DAI scores,spleen index,colon tissue pathology scores,and levels of TNF-αand IL-6 in serum(P<0.05,P<0.01),increased PI3K,phosphorylated Akt(p-Akt),and B-cell lymphoma-2(Bcl-2)-associated X protein(Bax)expression in colon tissue(P<0.05,P<0.01),and decreased serum IL-10 levels and colon tissue Bcl-2 protein expression(P<0.01).Compared with the model group,the ZJW groups showed significant improvement in UC symptoms,relieved colon tissue pathological damage,downregulated levels of inflammatory cytokines TNF-αand IL-6 in serum(P<0.01),inhibited expression of PI3K,p-Akt,and Bax proteins in colon tissue(P<0.05,P<0.01),and increased serum IL-10 levels and colon tissue Bcl-2 protein expression(P<0.01),with the high-dose group showing the best effect.Conclusion:ZJW effectively alleviates DSS-induced UC,and its mechanism may be related to the inhibition of the PI3K/Akt signaling pathway and regulation of apoptosis-related protein expression.