Effect of Bazi Bushen Capsules on Delaying Aging Process of Naturally Aging Mice Based on Immune-inflammation-aging

SONG Yahui; MA Kun; ZHANG Yaping; WANG Dandong; MAO Xinjing; ZHANG Shaolan; WEI Cong

doi:10.13422/j.cnki.syfjx.20230242

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Effect of Bazi Bushen Capsules on Delaying Aging Process of Naturally Aging Mice Based on Immune-inflammation-aging

更新时间：2023-03-30

- Effect of Bazi Bushen Capsules on Delaying Aging Process of Naturally Aging Mice Based on Immune-inflammation-aging
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 9, Pages: 146-155(2023)
- 作者机构：
  
  1.河北中医学院，石家庄 050091
  2.河北医科大学，石家庄 050017
  3.南京中医药大学，南京 210046
  4.络病研究与创新中药国家重点实验室，石家庄 050035
  5.河北省络病重点实验室，石家庄 050035
  6.国家中医药管理局中医络病学重点学科，石家庄 050035
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20230242
  CLC： R2-0;R33;R289;R339.3+8
- Published：05 May 2023，
  
  Published Online：10 February 2023，
  
  Received：30 December 2022，
- 稿件说明：
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宋亚辉,马坤,张雅萍等.基于免疫-炎症-衰老探讨八子补肾胶囊对自然衰老小鼠老化过程的延缓作用[J].中国实验方剂学杂志,2023,29(09):146-155.

SONG Yahui,MA Kun,ZHANG Yaping,et al.Effect of Bazi Bushen Capsules on Delaying Aging Process of Naturally Aging Mice Based on Immune-inflammation-aging[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(09):146-155.
宋亚辉,马坤,张雅萍等.基于免疫-炎症-衰老探讨八子补肾胶囊对自然衰老小鼠老化过程的延缓作用[J].中国实验方剂学杂志,2023,29(09):146-155. DOI： 10.13422/j.cnki.syfjx.20230242.

SONG Yahui,MA Kun,ZHANG Yaping,et al.Effect of Bazi Bushen Capsules on Delaying Aging Process of Naturally Aging Mice Based on Immune-inflammation-aging[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(09):146-155. DOI： 10.13422/j.cnki.syfjx.20230242.

摘要

目的

探讨八子补肾胶囊延缓自然衰老小鼠老化过程的功效作用及作用机制。

方法

以13月龄自然衰老小鼠为实验研究对象，将其按体质量随机分为4组，即自然衰老组，八子补肾低、高剂量组（1、2 g·kg

-1

），雷帕霉素组（0.002 g·kg

-1

）。通过行为学实验检测小鼠的衰弱表征并测定其虚弱指数；流式细胞术检测小鼠脾脏T和B淋巴细胞、效应性T细胞（TE）、记忆性T细胞（TM）、幼稚性T细胞（TN）、辅助性T细胞（Th）、细胞毒性T细胞（Tc）、Th1细胞、Th2细胞和调节性T细胞（Treg）比例；细胞增殖与活性检测（CCK-8）试剂盒检测小鼠淋巴细胞的增殖能力；苏木素-伊红（HE）染色观察小鼠脾脏组织病理学变化；免疫组化法（IHC）检测细胞周期蛋白依赖性激酶抑制剂2A（p16）和细胞周期蛋白依赖性激酶抑制剂1A（p21）的表达情况；实时荧光定量聚合酶链式反应（Real-time PCR）检测衰老相关蛋白p16和p21的mRNA表达情况；Luminex检测小鼠外周血血清炎症因子：肿瘤坏死因子-

（TNF-

）、

干扰素（IFN-

）、白细胞介素（IL）-1

、IL-2、IL-4、IL-10、IL-12 p70水平。

结果

与自然衰老组比较，八子补肾组和雷帕霉素组小鼠的衰弱表征明显改善，虚弱指数评分明显降低（

0.05，

0.01）；脾脏中T细胞、TN细胞、Tc细胞、Th2细胞和Treg细胞比例升高，TE细胞、TM细胞、Th细胞和Th1细胞比例下降（

0.05，

0.01）；脾淋巴细胞增殖能力增强（

0.05，

0.01）；脾脏红髓和白髓边缘区结构清晰，白髓面积增大，红髓面积相应减小；脾脏中衰老相关蛋白p16和p21蛋白表达量减少（

0.01）；p16和p21的mRNA表达量降低（

0.01）；血清中促炎细胞因子IL-1

、IL-2、IL-12 p70、IFN-

和TNF-

水平降低，抗炎因子IL-4和IL-10水平升高（

0.05，

0.01）。

结论

八子补肾胶囊具有调节自然衰老小鼠衰弱表征，调节机体免疫稳态及炎症水平，减少免疫细胞衰老的作用，起到了延缓衰老的作用功效。

Abstract

Objective

To study the effect of Bazi Bushen capsules on delaying the aging process of naturally aging mice and its mechanism.

Method

The mice were randomly divided into four groups according to their body weight， namely， aging group， low-dose Bazi Bushen capsules group （1 g·kg

-1

）， high-dose Bazi Bushen capsules group （2 g·kg

-1

）， and rapamycin group （0.002 g·kg

-1

）. The debilitating signs were detected by behavioral tests and the weakness index was measured. The percentages of spleen T and B lymphocytes， effector T cells （TE）， memory T cells （TM）， naive T cells （TN）， helper T cells （Th）， cytotoxic T cells （Tc），Th1 cells， Th2 cells， and regulatory T cells （Treg） were determined by flow cytometry. Cell proliferation and the cell counting kit-8 （CCK-8） assay was used to detect the proliferation of lymphocytes in mice. The hematoxylin-eosin （HE） staining was used to observe the histopathological changes in the mouse spleen. The expression of cyclin-dependent kinase inhibitor 2A （p16） and cyclin-dependent kinase inhibitor 1A （p21） was detected by immunohistochemistry （IHC）. The mRNA expression of senescence-related proteins p16 and p21 was determined by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and the inflammatory cytokines， including tumor necrosis factor-

（TNF-

）， interferon-

（IFN-

）， interleukin （IL）-1

， IL-2， IL-4， IL-10， and IL-12 p70， in peripheral blood of mice were detected by Luminex.

Result

As compared with the aging group， mice in the Bazi Bushen capsules and rapamycin groups showed significantly improved debilitating signs and reduced weakness index scores （

0.05，

0.01）， increased proportions of T cells， TN cells， Tc cells， Th2 cells， and Treg cells in the spleen， decreased proportions of TE cells， TM cells， Th cells， Th1 cells （

0.05，

0.01）， and increased proliferation of splenic lymphocytes （

0.05，

0.01）. In the Bazi Bushen capsules and rapamycin groups， clear structure of the red and white marrow marginal zone was observed in the spleen of mice， the area of the white marrow was increased， and the area of the red marrow was correspondingly decreased. The protein and mRNA expression of aging-related proteins p16 and p21 in the spleen was decreased （

0.01）， the levels of serum pro-inflammatory cytokines IL-1

， IL-2， IL-12 p70， IFN-

， and TNF-

levels were decreased， and the levels of IL-4 and IL-10 were increased （

0.05，

0.01） in the Bazi Bushen capsules and rapamycin groups as compared with the aging group.

Conclusion

Bazi Bushen capsules have the effect of regulating the debilitating signs of natural aging mice， regulating the immune homeostasis and inflammation level of the body， and reducing cell aging.

关键词

八子补肾胶囊炎症免疫衰老淋巴细胞亚群衰老相关蛋白

Keywords

Bazi Bushen capsulesinflammationimmunityaginglymphocyte subsetsaging-related proteins

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