目的 探讨左归降糖舒心方对糖尿病动脉粥样硬化小鼠肠道黏膜屏障的影响及机制。方法 Apoe -/- 雄性 小鼠适应性饲养 1 周后，给予小剂量链脲佐菌素（40 mg·kg -1 ）腹腔注射，每日 1 次，连续 5 d；检测小鼠空腹血 糖＞11.1 mmol·L -1 后，继续以高脂饲料喂养 12 周。造模结束后，按照随机数字表法将造模小鼠分为模型组、 二甲双胍组（0.065 g·kg -1 ）、左归降糖舒心方组（28.86 g·kg -1 ），另取 C57BL/6 雄性小鼠作为空白对照组，每组 5 只。各给药组按照上述剂量灌胃给药（15 mL·kg -1 ），空白对照组及模型组给予等量蒸馏水灌胃，每天 1 次， 连续 8 周。采用 ELISA 法检测小鼠血清中胰高血糖素样肽 1（GLP-1）、胆囊收缩素（CCK）、脂多糖（LPS）、白 细胞介素 18（IL-18）、白细胞介素 1β（IL-1β）的含量；苏木精-伊红（HE）染色法观察主动脉、回肠组织病理变 化；过碘酸雪夫（PAS）染色法观察回肠组织杯状细胞数量；RT-qPCR 法检测小鼠回肠组织肠道紧密连接蛋白 1 （Claudin-1）、闭锁小带蛋白 1（ZO-1）、咬合蛋白（Occludin）mRNA 表达情况；Western Blot 法检测回肠组织 NOD 样受体热蛋白结构域相关蛋白 3（NLRP3）、IL-1β 及 Gasdermin D-N 端片段（GSDMD-N）蛋白表达水平。结果 与 空白对照组比较，模型组小鼠血清 GLP-1、CCK 含量显著降低（P＜0.01）；血清 LPS、IL-1β、IL-18 含量均显 著升高（P＜0.01）；主动脉血管内膜增厚且结构紊乱，内壁不平整，存在大面积粥样斑块突出管腔，高倍镜下 可见斑块内胆固醇结晶，平滑肌细胞排列紊乱，内膜下可见泡沫细胞及脂质堆积，炎症细胞浸润；回肠组织结 构完整性被破坏，肠黏膜上皮排列紊乱，部分发生固有层增厚、中断，肠绒毛存在疏松断裂；回肠绒毛高度及 隐窝深度显著降低（P＜0.01）；回肠 PAS 阳性细胞表达量显著降低（P＜0.01）；回肠组织 Claudin-1、ZO-1、 Occludin mRNA 表达显著下调（P＜0.01）；回肠组织 NLRP3、IL-1β、GSDMD-N 蛋白表达水平均显著升高（P＜ 0.01）。与模型组比较，左归降糖舒心方组及二甲双胍组小鼠血清 GLP-1、CCK 含量显著升高（P＜0.01）；血清 LPS、IL-1β、IL-18 含量均显著降低（P＜0.01）；主动脉管腔狭窄程度及斑块病变减轻，血管内壁欠光滑，散 在粥样斑块，脂质蓄积及炎症细胞浸润情况均有不同程度改善；回肠黏膜上皮损伤均有所改善，回肠结构较完 整，肠绒毛密度增加，绒毛断裂状态减少；肠绒毛高度及隐窝深度明显增加（P＜0.05，P＜0.01）；回肠 PAS 阳 性细胞表达量明显增加（P＜0.05，P＜0.01）；回肠组织 Claudin-1、ZO-1、Occludin mRNA 表达明显上调（P＜ 0.05，P＜0.01）；回肠组织 NLRP3、IL-1β、GSDMD-N 蛋白表达水平均明显降低（P＜0.05，P＜0.01）。 结论 左归降糖舒心方可以改善糖尿病动脉粥样硬化小鼠的胃肠激素分泌，降低炎症因子水平，修复肠道黏 膜屏障，其作用机制可能与调控 NLRP3 炎症小体信号通路，抑制肠道炎症因子表达有关。

Objective To investigate the effect and mechanism of Zuogui Jiangtang Shuxin Formula （ZGJTSXF） on the intestinal mucosal barrier in mice with diabetic atherosclerosis. Methods After one week of acclimatization， male Apoe -/- mice were intraperitoneally injected with a low dose of streptozotocin （40 mg·kg -1 ） once daily for five consecutive days. After confirming fasting blood glucose levels ＞11.1 mmol·L -1 ，the mice were continuously fed a high-fat diet for 12 weeks. Upon successful modeling，the mice were randomly divided into the model group，the metformin group （0.065 g·kg -1 ），the ZGJTSXF group （28.86 g·kg -1 ），and male C57BL/6 mice served as the blank control group with five mice per group. The treatment groups received the respective drugs by gavage （15 mL·kg -1 ），while the blank control and model groups received an equal volume of distilled water，once daily for 8 consecutive weeks. Serum levels of glucagon-like peptide-1 （GLP-1），cholecystokinin （CCK），lipopolysaccharide （LPS），interleukin-18 （IL-18）， and interleukin-1β （IL-1β） were measured by ELISA. Pathological changes in the aorta and ileum were observed by hematoxylin-eosin （HE） staining. The number of goblet cells in ileal tissue was assessed by periodic acid-Schiff （PAS） staining. The mRNA expression levels of intestinal tight junction proteins Claudin-1，zonula occludens-1 （ZO-1）， and Occludin in the ileum were detected by RT-qPCR. The protein expression levels of NOD-like receptor thermal protein domain associated protein 3 （NLRP3），IL-1β，and Gasdermin D-N-terminal fragment （GSDMD-N） in the ileum were measured by Western Blot. Results Compared with the blank control group， the model group showed significantly decreased serum levels of GLP-1 and CCK （P＜0.01），and significantly increased serum levels of LPS， IL-1β，and IL-18 （P＜0.01）. The aortic intima was thickened and structurally disordered，with an uneven inner wall and large atherosclerotic plaques protruding into the lumen. Under high magnification，cholesterol crystals within the plaques，disordered smooth muscle cell arrangement，subintimal foam cells，lipid accumulation，and inflammatory cell infiltration were observed. The structural integrity of the ileum was disrupted，with disordered arrangement of the intestinal mucosal epithelium， partial thickening and interruption of the lamina propria， and loose and broken intestinal villi. The ileal villus height and crypt depth were significantly reduced （P＜0.01）. The number of PAS- positive cells in the ileum was significantly decreased （P＜0.01）. The mRNA expression levels of Claudin-1，ZO-1， and Occludin in the ileum were significantly downregulated （P＜0.01）. The protein expression levels of NLRP3，IL-1β， and GSDMD-N in the ileum were significantly increased （P＜0.01）. Compared with the model group，the ZGJTSXF group and the metformin group showed significantly increased serum levels of GLP-1 and CCK （P＜0.01）， and significantly decreased serum levels of LPS，IL-1β，and IL-18 （P＜0.01）. The degree of aortic lumen stenosis and plaque lesions was reduced，with a relatively smoother vascular wall，scattered atherosclerotic plaques，and varying degrees of improvement in lipid accumulation and inflammatory cell infiltration. Damage to the ileal mucosal epithelium was ameliorated，the ileal structure was relatively intact，intestinal villus density increased，and villus breakage was reduced. The ileal villus height and crypt depth were significantly increased （P＜0.05，P＜0.01）. The number of PAS- positive cells in the ileum was significantly increased （P＜0.05，P＜0.01）. The mRNA expression levels of Claudin-1， ZO-1，and Occludin in the ileum were significantly upregulated （P＜0.05，P＜0.01）. The protein expression levels of NLRP3， IL-1β， and GSDMD-N in the ileum were significantly decreased （P＜0.05， P＜0.01）. Conclusion ZGJTSXF can improve gastrointestinal hormone secretion，reduce inflammatory factor levels，and repair the intestinal mucosal barrier in mice with diabetic atherosclerosis. Its mechanism of action may be related to the regulation of the NLRP3 inflammasome signaling pathway and the inhibition of intestinal inflammatory factor expression.

R285.5

国家自然科学基金项目（82205077）；湖南省自然科学基金项目（2022JJ40315）；湖南省教育厅资助科研项目（21B0390）；湖南省卫生 健康委科研计划项目（D202303067601）；中药粉体与创新药物省部共建国家重点实验室培育基地开放基金项目（21PTKF1016）。