归脾丸对D-半乳糖致衰老小鼠的免疫调节作用研究
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篇名: 归脾丸对D-半乳糖致衰老小鼠的免疫调节作用研究
TITLE: Study on immunomodulatory effect of Guipi pills on D-galactose-induced aging mice
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400mg/kgD-gal以复制衰老模型;CON组与MOD组灌胃蒸馏水,POS组灌胃300mg/kg匹多莫德口服溶液,GD组和GG组分别灌胃300mg/kg和600mg/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。
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 TNF-α, PI3K and p-Akt.
期刊: 2023年第34卷第12期
作者: 李思晓;苑广信;马月;王宇桐
AUTHORS: LI Sixiao,YUAN Guangxin,MA Yue,WANG Yutong
关键字: 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
KEYWORDS: Guipi pills; immuno-modulatory; network phar-
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