Page 24 - 《中国药房》2023年12期

归脾丸对D-半乳糖致衰老小鼠的免疫调节作用研究 Δ

李思晓 ，苑广信 ，马 月，王宇桐（北华大学药学院，吉林 吉林 132013）
#
＊
中图分类号 R965 文献标志码 A 文章编号 1001-0408（2023）12-1426-05
DOI 10.6039/j.issn.1001-0408.2023.12.04

摘 要 目的 研究归脾丸对D-半乳糖（D-gal）致衰老小鼠的免疫调节作用。方法 运用网络药理学方法筛选归脾丸发挥免疫作
用的相关靶点和相关通路，并通过药效学实验进行验证。将105只雄性ICR小鼠随机分为空白对照（CON）组、模型对照（MOD）
组、阳性对照（POS）组、归脾丸低剂量（GD）组和归脾丸高剂量（GG）组，除CON组皮下注射生理盐水外，其余各组小鼠皮下注射
400 mg/kg D-gal以复制衰老模型；CON组与MOD组灌胃蒸馏水，POS组灌胃300 mg/kg匹多莫德口服溶液，GD组和GG组分别灌
胃300 mg/kg和600 mg/kg的归脾丸水溶液，每日1次，共计8周。给药结束后，采集血清及脾脏组织，检测小鼠血清中白细胞介素
2（IL-2）、IL-4、IL-6、肿瘤坏死因子α（TNF-α）的含量以及免疫球蛋白G（IgG）、IgM和IgA含量；计算脾脏指数并观察脾脏组织病理
学改变；检测小鼠脾脏组织中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）、丙二醛（MDA）活性和8-羟基脱氧鸟苷（8-
OHdG）含量；检测除POS组外小鼠脾脏组织中TNF/磷酸肌醇-3-激酶-蛋白激酶B（PI3k-Akt）途径中相关蛋白的表达情况。结果
网络药理学分析结果显示，TNF、IL-6、Akt1为核心靶点。药效学研究结果显示，与MOD组比较，归脾丸各剂量组小鼠血清中IL-
2、IL-4、IgG、IgM、IgA含量均显著升高，TNF-α、IL-6含量均显著降低，脾脏指数均显著升高（P＜0.05或P＜0.01）；淋巴细胞弥漫性
增生的现象改善，脾细胞紧密排列，白髓和红髓之间界线清晰；脾脏组织中SOD、GSH-Px活性均显著升高，MDA活性和8-OHdG
含量均显著降低，TNF-α、PI3K、p-Akt蛋白表达水平均显著降低（P＜0.05或P＜0.01）。结论 归脾丸能够调节D-gal致衰老小鼠的
免疫功能，其作用可能与调控TNF/PI3k-Akt途径进而减轻小鼠脾脏组织氧化应激损伤以及调节TNF-α、PI3K、p-Akt蛋白水平的
表达有关。
关键词 归脾丸；免疫调节；网络药理学；药效学；肿瘤坏死因子；磷酸肌醇-3-激酶；蛋白激酶B

Study on immunomodulatory effect of Guipi pills on D-galactose-induced aging mice
LI Sixiao，YUAN Guangxin，MA Yue，WANG Yutong（School of Pharmacy， Beihua University， Jilin Jilin
132013， China）

ABSTRACT OBJECTIVE To study the immunomodulatory effect of Guipi pills on D-galactose（D-gal）-induced aging mice.
METHODS The immune-related targets and related pathways for Guipi pills to exert immune effects were screened by network
pharmacology and verified through pharmacodynamic experiments. Totally 105 male ICR mice were randomly divided into blank
control （CON） group， model control （MOD） group， positive control （POS） group， Guipi pills low-dose （GD） group and Guipi
pills high-dose （GG） group. Except for the CON group， other groups were subcutaneously injected with 400 mg/kg D-gal to induce
the aging model； CON group and MOD group were given distilled water， POS group was given 300 mg/kg pidotimod oral solution
intragastrically， GD group and GG group were given Guipi pills 300， 600 mg/kg intragastrically， once a day， for 8 weeks. After
medication， the serum and spleen were collected， and the contents of interleukin 2 （IL-2）， IL-4， IL-6 and tumor necrosis factor α
（TNF-α）， and the contents of immunoglobulin G （IgG）， IgM and IgA were detected. The spleen index was calculated and the
histopathological changes in the spleen were observed. The activities of superoxide dismutase （SOD）， glutathione peroxidase （GSH-
Px） and malondialdehyde （MDA）， and the content of 8-hydroxy-2 deoxyguanosine （8-OHdG） in spleen were detected； the
expression of TNF/phosphoinositide-3-kinase-threonine protein kinase （PI3k-Akt）-related proteins in spleen was detected except for
POS group. RESULTS The results of network pharmacology showed that TNF， IL-6 and Akt1 were core targets. The results of
pharmacodynamic study showed that compared with MOD group， the contents of IL-2， IL-4， IgG， IgM and IgA were increased
significantly in Guipi pills groups， while the contents of TNF- α and IL-6 were decreased significantly； the spleen index was
increased significantly （P＜0.05 or P＜0.01）. The phenomenon of diffuse proliferation of lymphocytes was improved， the spleen
cells were closely arranged， and the line between the white pulp and red pulp was clear. The activities of SOD and GSH-Px in
spleen were increased significantly， while the activity of MDA， the content of 8-OHdG， and the protein expressions of TNF-α，
PI3K and p-Akt were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS Guipi pills can regulate the immune function
of D-gal-induced aging mice， which is related to regulating the TNF/PI3k-Akt pathway， thereby reducing oxidative stress damage in
spleen tissue of mice， and regulating protein expressions of
Δ 基金项目 吉林省科技发展计划项目（No.YDZJ202201ZYTS197）
TNF-α， PI3K and p-Akt.
＊ 第一作者 硕 士 研 究 生 。 研 究 方 向 ：药 物 分 析 。 E-mail：
1073085922@qq.com KEYWORDS Guipi pills； immuno-modulatory； network phar-
# 通信作者 教授，硕士生导师，博士。研究方向：药品质量控制。 macology； pharmacodynamics； tumor necrosis factor；
E-mail：17287598@qq.com phosphoinositide-3-kinase； threonine protein kinase

· 1426 · China Pharmacy 2023 Vol. 34 No. 12 中国药房 2023年第34卷第12期