目的 基于“肝病实脾”理论探讨白术多糖对酒精性肝损伤（ALI）大鼠的保护作用。方法 将大鼠随机 分为模型组，白术多糖低、高剂量组，多烯磷脂酰胆碱组，白术多糖高剂量+氧化三甲胺组及对照组，每组 12 只。除对照组大鼠外，其余组大鼠均通过连续 6 周灌胃白酒的方式进行模型复制。模型复制的同时进行药 物干预，给药 1 日 1 次，持续 6 周。检测大鼠血清中谷丙转氨酶（ALT）及谷草转氨酶（AST）水平的变化；HE 染色法观察大鼠肝组织、脾脏组织的病理变化；检测大鼠脾脏指数；流式细胞术检测脾脏组织中 CD4+ 、CD8+ 及 CD4+ /CD8+ 表达；检测大鼠脾脏组织中丙二醛（MDA）、超氧化物歧化酶（SOD）含量及活性氧（ROS）荧光强度 的变化；Western Blot 法检测脾脏及肝脏组织中葡萄糖调节蛋白 78（GRP78）、半胱氨酸天冬氨酸蛋白酶 12 （Caspase-12）蛋白水平。结果 与对照组比较，模型组大鼠肝脏及脾脏组织病理损伤严重，血清中 AST、ALT 水平，脾脏指数，脾脏组织中 CD8+ 水平、MDA 含量、ROS 荧光强度以及脾脏、肝脏组织中 GRP78、Caspase-12 蛋白表达水平均升高，脾脏组织中 CD4+ 水平、CD4+ /CD8+ 、SOD 含量均降低（均 P＜0.05）；与模型组比较，白 术多糖低剂量组、白术多糖高剂量组、多烯磷脂酰胆碱组大鼠肝组织及脾脏组织病理损伤有所缓解，血清中 AST、ALT 水平，脾脏指数，脾脏组织中 CD8+水平、MDA 含量、ROS 荧光强度以及脾脏、肝脏组织中 GRP78、Caspase-12 蛋白表达均降低，脾脏组织中 CD4+ 水平、CD4+ /CD8+ 、SOD 含量均升高（均 P＜0.05）；氧 化三甲胺减弱了高剂量白术多糖对 ALI 大鼠脾脏氧化应激的抑制作用以及对免疫功能的改善作用（均 P＜0.05）。 结论 白术多糖可能通过抑制 ROS/GRP78/Caspase-12 信号通路抑制 ALI 大鼠脾脏氧化应激并改善免疫功能。

Objective To investigate the protective effect of polysaccharide from atractylodes on alcoholic liver injury （ALI） in rats based on the theory of “treating the liver by reinforcing the spleen”. Methods The rats were randomly divided into model group，polysaccharide from atractylodes low-dose and high-dose group，polyene phosphatidylcholine group，polysaccharide from atractylodes high-dose + trimethylamine oxide group and control group，with 12 rats in each group. Except for the control group，the model was replicated by intragastric administration of liquor for 6 consecutive weeks. The model was replicated while drug intervention was administered once a day for 6 consecutive weeks. The changes in serum alanine transaminase （ALT） and aspartate aminotransferase （AST） levels in rats were detected； HE staining was applied to detect pathological changes in rat liver and spleen tissues； the spleen index of rats was detected；flow cytometry was applied to detect the expressions of CD4+ ，CD8+ ，and CD4+ /CD8+ in splenic tissue；the changes in malondialdehyde （MDA），superoxide dismutase （SOD） contents，and reactive oxygen species （ROS） fluorescence intensity in rat spleen tissue were detected；Western Blot detect glucose regulated protein 78 （GRP78）， cysteinyl aspartate specific protease 12 （Caspase-12） proteins in spleen and liver tissues. Results Compared with the control group，the model group had severe pathological damage to the liver and spleen tissues of rats，the levels of AST and ALT in serum，spleen index，CD8+ level in spleen tissue，MDA content，ROS fluorescence intensity，and the protein expressions of GRP78 and Caspase-12 in spleen and liver tissues elevated，the levels of CD4+，CD4+ /CD8+， and SOD content in spleen tissue decreased （P＜0.05）；compared with the model group，the pathological damage to the liver and spleen tissues of rats in the polysaccharide from atractylodes low-dose group， polysaccharide from atractylodes high-dose group and polyene phosphatidylcholine group was alleviated， the levels of AST and ALT in serum， spleen index， CD8+ level in spleen tissue， MDA content， ROS fluorescence intensity， and the protein expressions of GRP78 and Caspase-12 in spleen and liver tissues reduced，the levels of CD4+ ，CD4+ /CD8+ ，and SOD content in spleen tissue increased （P＜0.05）； trimethylamine oxide weakened the inhibitory effect of high-dose polysaccharide from atractylodes on oxidative stress in the spleen of ALI rats and its improvement on immune function. Conclusion Polysaccharide from atractylodes may inhibit spleen oxidative stress and improve immune function in ALI rats by inhibiting ROS/GRP78/Caspase-12 signaling pathway.

R285.5

河南省中医药科学研究专项课题（2022ZY1147）。