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[参芪二皮颗粒对S_(180)荷瘤小鼠的免疫功能调节及抑瘤作用]

[Immune function regulation and tumor-suppressive effects of Shenqi Erpi Granules on S_(180) tumor-bearing mice].

作者信息

Zhang Xiong-Wei, Jiang Yan-Ning, Qi Hu, Li Bin, Gao Yuan-Lin, Zhang Ze-Yang, Feng Jian-An, Li Xi, Zeng Nan

机构信息

School of Pharmacy, Chengdu University of Traditional Chinese Medicine Chengdu 611137, China State Key Laboratory of Southwestern Chinese Medicine Resources, Chengdu University of Traditional Chinese Medicine Chengdu 611137, China.

Institute of Traditional Chinese Medicine, Sichuan Academy of Chinese Medicine Sciences Chengdu 610031, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2025 Jul;50(13):3753-3764. doi: 10.19540/j.cnki.cjcmm.20250507.401.

Abstract

This study aims to establish the S_(180) tumor-bearing mice model, and to investigate the influence of Shenqi Erpi Granules(SQEPG) on immune function, as well as the drug's tumor-suppressive effect and mechanism. SPF grade KM mice(half male and half female) were randomly divided into 6 groups: a control group, a model group, a cyclophosphamide group(50 mg·kg(-1)), as well as SQEPG groups in low-, medium-, and high-dose(5.25, 10.5, 21 g·kg(-1)). The control group and the model group were given distilled water, and the other 4 groups were given the corresponding drugs by gavage. The administration continued for 10 days before the mice were sacrificed. The antitumor and immune regulation effects of SQEPG were evaluated. The effect of SQEPG on delayed type hypersensitivity reaction(DTH), carbon clearance index, and serum hemolysin antibody level was observed to reflect the effect on the immune function of tumor-bearing mice. Tumor weight was recorded to calculate the tumor suppression rate and the immune organ index. Hematoxylin-eosin(HE) staining was used to detect morphological changes in tumor tissues. Flow cytometry was employed to detect the percentage of CD4+ and CD8+ T-cells in the spleen tissues and the tumor tissue apoptosis levels. Immunohistochemistry was conducted to detect the KI67 protein expression level of tumor tissues. ELISA resorted to the detection of the following expression levels in tumor tissues: tumor necrosis factor-α(TNF-α), interleukin-2(IL-2), interferon-γ(IFN-γ). Western blot was performed to detect the expression levels of caspase-3, B-cell lymphoma-2(Bcl-2), Bcl-2-associated X protein(Bax), cyclin-dependent kinases 4(CDK4), G_1/S-specific cyclin D1(cyclin D1), and vascular endothelial growth factor A(VEGFA). The results showed that, compared with the model group, the SQEPG could increase the swelling of the auricle of the tumor-bearing mice; significantly increase the phagocytic index of carbon granule contour(P<0.05 or P<0.01), and the middle dose of SQEPG could significantly increase the antibody level of hemolysin(P<0.05); different doses of SQEPG significantly inhibit the growth of the tumor, and decrease the mass of the tumor tissues(P<0.05 or P<0.01); the low dose of SQEPG significantly decreased spleen index(P<0.05), low and high doses of SQEPG increased thymus index, while medium doses of SQEPG decreased thymus index. High doses of SQEPG significantly elevated the levels of CD4+ and CD8+ T-cells in the spleens of the homozygous mice(P<0.01 or P<0.001), and increased the apoptosis rate of the cells of the tumor tissues(P<0.05); Meanwhile, high-dose SQEPG elevated the levels of immunity factors such as IL-2, IFN-γ and TNF-α in the serum of tumor-bearing mice(P<0.01); medium-and high-dose SQEPG significantly lowered the rate of positive expression of KI67 protein in tumor tissues(P<0.01). Compared with the model group, high-dose SQEPG significantly up-regulated the expression of caspase-3 and Bax proteins in tumor tissues(P<0.05), and significantly down-regulated the expression of CDK4, cyclin D1, and VEGFA proteins(P<0.05 or P<0.01). In conclusion, SQEPG has the effect of improving immune function and inhibiting tumor growth in tumor-bearing mice. Its mechanism of tumor-suppressive effects may be related to apoptosis promotion, cell cycle progression block, and tumor cell proliferation inhibition.

摘要

本研究旨在建立S_(180)荷瘤小鼠模型,探讨参芪二皮颗粒(SQEPG)对免疫功能的影响以及该药物的抑瘤作用和机制。将SPF级KM小鼠(雌雄各半)随机分为6组:对照组、模型组、环磷酰胺组(50 mg·kg(-1))以及低、中、高剂量的SQEPG组(5.25、10.5、21 g·kg(-1))。对照组和模型组给予蒸馏水,其他4组通过灌胃给予相应药物。给药持续10天,然后处死小鼠。评估SQEPG的抗肿瘤和免疫调节作用。观察SQEPG对迟发型超敏反应(DTH)、碳清除指数和血清溶血素抗体水平的影响,以反映其对荷瘤小鼠免疫功能的影响。记录肿瘤重量以计算肿瘤抑制率和免疫器官指数。采用苏木精-伊红(HE)染色检测肿瘤组织的形态学变化。运用流式细胞术检测脾脏组织中CD4+和CD8+ T细胞的百分比以及肿瘤组织凋亡水平。进行免疫组织化学检测肿瘤组织中KI67蛋白表达水平。采用ELISA检测肿瘤组织中以下表达水平:肿瘤坏死因子-α(TNF-α)、白细胞介素-2(IL-2)、干扰素-γ(IFN-γ)。进行蛋白质免疫印迹法检测半胱天冬酶-3、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、细胞周期蛋白依赖性激酶4(CDK4)、G_1/S特异性细胞周期蛋白D1(细胞周期蛋白D1)和血管内皮生长因子A(VEGFA)的表达水平。结果显示,与模型组相比,SQEPG可增加荷瘤小鼠耳廓肿胀度;显著提高碳粒廓清吞噬指数(P<0.05或P<0.01),且中剂量的SQEPG可显著提高溶血素抗体水平(P<0.05);不同剂量的SQEPG均显著抑制肿瘤生长,降低肿瘤组织质量(P<0.05或P<0.01);低剂量的SQEPG显著降低脾脏指数(P<0.05),低、高剂量的SQEPG可提高胸腺指数,而中剂量的SQEPG降低胸腺指数。高剂量的SQEPG显著提高纯合小鼠脾脏中CD4+和CD8+ T细胞水平(P<0.01或P<0.001),并提高肿瘤组织细胞凋亡率(P<0.05);同时,高剂量的SQEPG提高荷瘤小鼠血清中IL-2、IFN-γ和TNF-α等免疫因子水平(P<0.01);中、高剂量的SQEPG显著降低肿瘤组织中KI67蛋白阳性表达率(P<0.01)。与模型组相比,高剂量的SQEPG显著上调肿瘤组织中半胱天冬酶-3和Bax蛋白的表达(P<0.05),并显著下调CDK4、细胞周期蛋白D1和VEGFA蛋白的表达(P<0.05或P<0.01)。综上所述,SQEPG具有改善荷瘤小鼠免疫功能和抑制肿瘤生长的作用。其抑瘤作用机制可能与促进细胞凋亡、阻滞细胞周期进程以及抑制肿瘤细胞增殖有关。

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