基于Th1、Th2细胞因子探讨清肠合剂改善术后腹腔粘连的机制

Mechanism study of Qingchang mixture in the treatment of postoperative abdominal adhesions by regulating the expression of Th1 and Th2 cytokines

目的探讨清肠合剂改善大鼠术后腹腔粘连的效果及其可能的作用机制。方法75只SD大鼠随机分为5组:空白组、假手术组、模型组、清肠合剂组及透明质酸钠组。除空白组和假手术组外,其余3组采用盲肠刮擦法建立腹腔粘连大鼠模型;透明质酸钠组用2 mL透明质酸钠凝胶均匀涂抹受损的腹壁及盲肠后关腹。清肠合剂组造模成功后每日予2 mL清肠合剂(14.58 g/kg)灌胃1次,其余4组予等体积生理盐水灌胃。每组分别于术后第3、7、14天处死5只大鼠,采用Nair’s评分评估腹腔粘连情况,并收集术后7 d粘连组织或正常腹膜组织,应用荧光定量PCR法检测白细胞介素(IL)-4、信号转导和转录激活因子6(STAT-6)及干扰素-γ(IFN-γ)的mRNA表达水平;Western blot检测IL-4、IL-10、STAT-6及IFN-γ的蛋白表达情况;苏木精-伊红染色后光镜下观察各组粘连组织病理变化,评估炎症和纤维形成情况。结果与空白组和假手术组相比,模型组粘连组织中IL-4、STAT-6的mRNA和蛋白表达水平以及IL-10的蛋白表达水平下降,IFN-γ的mRNA和蛋白表达水平、炎症评分及纤维评分均升高(P<0.05)。与模型组相比,清肠合剂组和透明质酸钠组的IL-4和STAT-6的mRNA和蛋白表达水平以及IL-10蛋白表达水平均升高;Nair’s评分、炎症评分、纤维评分以及IFN-γ的mRNA和蛋白表达水平均降低(P<0.05)。结论清肠合剂可以抑制术后腹腔粘连的形成,其机制可能与调控辅助性T细胞1、辅助性T细胞2相关的细胞因子,降低炎症反应水平有关。

Objective To elucidate the efficacy and possible mechanism of Qingchang mixture in ameliorating postoperative abdominal adhesions in rats.Methods Seventy-five Sprague-Dawley rats were randomly allocated into five experimental groups:the control group,the sham-operated group,the model group,the Qingchang mixture treatment group and the sodium hyaluronate treatment group.Except the control group and the sham-operated group,the other three groups were treated with cecal abrasion method to establish the rat model of abdominal adhesion.In the sodium hyaluronate group,2 mL sodium hyaluronate gel was meticulously applied to the injured abdominal wall and cecum prior to abdominal closure.Following successful establishment of adhesion model,the Qingchang mixture group received a daily oral gavage of 2 mL Qingchang mixture(14.58 g/kg),while the other four groups were given equal volume normal saline administration.In each group,five rats were euthanized on postoperative days 3,7 and 14 to assess abdominal adhesions using Nair’s scoring system.Adhesive tissue or normal peritoneal tissue were harvested on postoperative day 7,and mRNA expression levels of interleukin-4(IL-4),signal transducer and activator of transcription 6(STAT-6)and interferon-γ(IFN-γ)were quantified via fluorescence-based real-time PCR.Concurrently,Western blot analysis was employed to evaluate protein expression levels of IL-4,interleukin-10(IL-10),STAT-6 and IFN-γ.Pathological alterations in adhesive tissue were visualized using hematoxylin-eosin(HE)staining under light microscopy,and inflammation and fibrotic changes were assessed accordingly.Results Compared with the blank and sham-operated groups,mRNA and protein expression levels of IL-4 and STAT-6 were significantly downregulated in the model group,protein expression level of IL-10 was also reduced.Conversely,the mRNA and protein expression levels of IFN-γ,as well as the inflammation and fibrosis scores were significantly elevated(P<0.05).In comparison to the model group,IL-4 and STAT-6 mRNA and protein expression levels were increased in the Qingchang mixture group and the hyaluronic acid group,along with an increase in IL-10 protein expression.Conversely,these groups exhibited a significant reduction in Nair’s scores,inflammation scores,fibrosis scores,and IFN-γmRNA and protein expression levels were decreased(P<0.05).Conclusion Qingchang mixture appears to suppress the development of postoperative peritoneal adhesions,likely through mechanism that involves modulating the expression of T-helper 1 and T-helper 2 cytokines,thereby attenuating inflammatory response.