Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2010, Vol. 8 ›› Issue (11): 1053-1059.doi: 10.3736/jcim20101109

• Original Expreimental Research • Previous Articles     Next Articles

Effects of Chinese herbal medicine Bushen Gubiao Recipe on toll-like receptor 4 and CD4 +CD25 +foxp3 +regulatory T cells in mice with recurrent respiratory tract infections

Ya-bing Zhoua,Jian-er Yub, Jie Wub,Li Baib, Li-li Huob, Xin-guang Zhangb,Li-qing Lib   

  1. a Department of Traditional Chinese Medicine, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
    b Department of Paediatrics, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200071, China
  • Received:2010-07-18 Accepted:2010-08-13 Online:2010-11-20 Published:2010-11-15
  • Contact: Jian-er Yu


To evaluate the effects of Bushen Gubiao Recipe (BGR), a compound traditional Chinese herbal medicine, on toll-like receptor 4 (TLR4) signaling pathway and CD4 +CD25 +foxp3 +regulatory T cells (CD4 +CD25 +foxp3 +Tregs) in mice with recurrent respiratory tract infections (RRTIs).


A mouse model of kidney-yang deficiency which simulated physical characteristics of RRTIs was established by intraperitoneal injection of hydrocortisone for 14 d. The model mice were divided into 4 groups, model group, high-dose BGR group, low-dose BGR group, and nucleic acid and casein oral solution group. They were administered respectively with distilled water, high-dose BGR (50 g/kg body weight), low-dose BGR (25 g/kg body weight) and nucleic acid and casein oral solution. Besides, a normal control group was set up and gastrogavaged with distilled water. The effect of intervention was evaluated 4 weeks later by estimating the changes in behaviors of mice. Expressions of TLR4 and nuclear factor-kappa B (NF-κB) p65 in lung tissue were detected by immunohistochemical SABC method, the expression of TLR4 mRNA in lung tissue was detected by fluorescence quantitative-polymerase chain reaction (FQ-PCR), and the level of blood CD4 +CD25 +foxp3 + Tregs was determined by flow cytometry.


A kidney-yang deficiency mouse model with RRTIs was successfully established by intraperitoneal injection of hydrocortisone. BGR could improve the abnormal behavioral condition of the mice and enhance the expressions of TLR4 and NF-κB p65 in the lung tissue. Expression of TLR4 mRNA in high-dose BGR group was higher than that in model group (P<0.05), while the difference was not statistically significant between high-dose BGR group and low-dose BGR group (P>0.05). Levels of CD4 +CD25 +foxp3 +Tregs in high-dose BGR group and nucleic acid and casein oral solution group were lower than that in model group (P<0.05), while the difference was not statistically significant between high-dose BGR group and low-dose BGR group (P>0.05).


BGR can improve the behavior of the kidney-yang deficiency mice, and improve the innate immune function by up-regulating TLR/NF-κB signaling pathway. BGR can adjust the immune imbalance of T-helper cell (Th) 1/Th2 through reducing the activity of CD4 + CD25 +foxp3 +Tregs and enhancing the immune response of Th1 type.

Key words: Chinese herbal drugs, Bushen Gubiao Recipe, Respiratory tract infections, Toll-like receptor 4, T-lymphocytes, regulatory, Nuclear factor-kappa B, Mice

Figure 1

Expression of TLR4 in lung tissue of mice assayed by immunohistochemical method with DAB staining in different groups (Light microscopy, ×250) A: Normal; B: Model; C: High-dose BGR; D: Low-dose BGR; E: Nucleic acid and casein oral solution."

Table 1

Expressions of TLR4 and NF-κB p65 in lung tissue of mice assayed by immunohistochemical staining in different groups"

Figure 2

Expression of NF-κB p65 in lung tissue of mice assayed by immunohistochemical method with DAB staining in different groups (Light microscopy, ×250) A: Normal; B: Model; C: High-dose BGR; D: Low-dose BGR; E: Nucleic acid and casein oral solution."

Table 2

Expression of TLR4 mRNA in lung tissue of mice messured with real-time FQ-PCR in different groups"

Table 3

CD4+CD25+foxp3+Tregs in peripheral blood of mice determined by flow cytometry in different groups"

[1] Zhao SY . Progress in the clinical research into respiratory diseases in children[J]. Zhongguo Shi Yong Er Ke Za Zhi, 2009,24(5):335-337
doi: 10.3969/j.issn.1007-614x.2013.03.001
赵顺英 . 儿童呼吸系统疾病临床研究进展[J]. 中国实用儿科杂志, 2009,24(5):335-337
doi: 10.3969/j.issn.1007-614x.2013.03.001
[2] Yu JE, Huo LL, Wu M, Lu BL, Zhang XF, Guan Y . The influence of Bushen Gubiao Decoction on T Cell subpopulation, immunoglobulin, natural killer lymphocyte and cortisol in children of recurrent respiratory tract infections[J]. Hunan Zhong Yi Za Zhi, 2006,22(5):3-5 5
虞坚尔, 霍莉莉, 吴敏, 卢伯良, 张晓峰, 管宇 .补肾固表方对复感儿T细胞亚群﹑免疫球蛋白﹑NK﹑皮质醇的影响[J].湖南中医杂志, 2006, 22(5):3-
[3] Yu JE, Huo LL, Li L, Li BS, Chen FF . Regulative effects of Bushengubiao Recipe on unbalanced expressions of Th1 and Th2 cytokines from peripheral blood in children with recurrent respiratory tract infections[J]. Beijing Zhong Yi Yao Da Xue Xue Bao, 2007,30(9):630-633
doi: 10.3321/j.issn:1006-2157.2007.09.017
虞坚尔, 霍莉莉, 李莉, 李本尚, 陈凡凡 . 补肾固表方对复感儿外周血Th1和Th2型细胞因子表达失衡的调节作用[J]. 北京中医药大学学报, 2007,30(9):630-633
doi: 10.3321/j.issn:1006-2157.2007.09.017
[4] Yu JE, Huo LL, Wu M, Lu WW . Influence of Bushengubiao Prescription on IFN-γ and IL-4 protein content in culture-supernatant of Th cells in patients with RRTI[J]. Zhonghua Zhong Yi Yao Za Zhi, 2007,22(7):451-453
doi: 10.3969/j.issn.1673-1727.2007.07.009
虞坚尔, 霍莉莉, 吴敏, 路薇薇 . 补肾固表方对复感儿Th细胞培养上清液中γ-干扰素与白细胞介素-4蛋白含量的影响[J]. 中华中医药杂志, 2007,22(7):451-453
doi: 10.3969/j.issn.1673-1727.2007.07.009
[5] Wu L, Cui MX, Xia XQ . Changes in immune function of children patients with repeated respiratory infection and effect of treatment with nucleotide and casein injection[J]. Zhongguo Lin Chuang Yao Li Xue Yu Zhi Liao Xue, 2006,11(4):467-469
doi: 10.3969/j.issn.1009-2501.2006.04.025
吴琳, 崔敏娴, 夏晓青 . 反复呼吸道感染患儿免疫功能变化及核酪注射治疗疗效观察[J]. 中国临床药理学与治疗学, 2006,11(4):467-469
doi: 10.3969/j.issn.1009-2501.2006.04.025
[6] Wang YY . New strategy for treatment of respiratory infection and the predominance of traditional Chinese medicine[J]. J Chin Integr Med, 2004,2(3):167-171
王轶宇 . 肺部感染治疗新对策及中医药治疗的优势[J]. 中西医结合学报, 2004,2(3):167-171
[7] Chen Q. Methodology of research in pharmacology of traditional Chinese medicine(2nd ed)[M]. Beijing: People’s Medical Publishing House, 2006,( 2nd ed):1049
陈奇 . 中药药理研究方法学(第二版)[M]. 北京: 人民卫生出版社, 2006,( 第二版):1049
[8] Xiao J, Wang YX, Gao JD . Progress in study of kidney-yang deficiency syndrome[J]. Shanghai Zhong Yi Yao Da Xue Xue Bao, 2008,22(2):73-77
肖静, 王毅兴, 高建东 . 肾阳虚证的研究进展[J]. 上海中医药大学学报, 2008,22(2):73-77
[9] Yu JE, Chen MH, Li HG, Zhao J . Influence of “kidney-tonifying and exterior-consolidating formula” on immunity of mouse model with deficiency of lung and kidney[J]. Shanghai Zhong Yi Yao Da Xue Xue Bao, 2000,14(3):35-37
虞坚尔, 陈慕涵, 李合国, 赵鋆 . 补肾固表方对肺肾两虚小鼠模型免疫功能的影响[J]. 上海中医药大学学报, 2000,14(3):35-37
[10] Orange JS, Levy O, Geha RS . Human disease resulting from gene mutations that interfere with appropriate nuclear factor-kappaB activation[J]. Immunol Rev, 2005,203(1):21-37
doi: 10.1111/imr.2005.203.issue-1
[11] Peng JH, Fang ZH, Cui JW, Feng Q, Xu LL, Gu HT, Hu YY . Effects of Jianpi Huoxue Decoction on Kupffer cell signal pathway activation in rats with liver injury induced by Lieber-Decarli liquid diet and lipopolysaccharide[J]. J Chin Integr Med, 2007,5(3):302-306
彭景华, 方志红, 崔剑巍, 冯琴, 许丽莉, 顾宏图, 胡义扬 . 健脾活血方对酒精复合内毒素脂多糖诱导的肝损伤大鼠库普弗细胞活化信号通路的干预[J]. 中西医结合学报, 2007,5(3):302-306
[12] Rozkova D, Horvath R, Bartunkova J, Spisek R . Glucocorticoids severely impair differentiation and antigen presenting function of dendritic cells despite upregulation of Toll-like receptors[J]. Clin Immunol, 2006,120(3):260-271
doi: 10.1016/j.clim.2006.04.567
[13] Zhang X, Shan P, Qureshi S, Homer R, Medzhitov R, Noble PW, Lee PJ . Cutting edge: TLR4 deficiency confers susceptibility to lethal oxidant lung injury[J]. J Immunol, 2005,175(8):4834-4838
doi: 10.4049/jimmunol.175.8.4834 pmid: 16210584
[14] Hauber HP, Tulic MK, Tsicopoulos A, Wallaert B, Olivenstein R, Daigneault P, Hamid Q . Toll-like receptors 4 and 2 expression in the bronchial mucosa of patients with cystic fibrosis[J]. Can Respir J, 2005,12(1):13-18
doi: 10.1155/2005/648984
[15] Cole LE, Elkins KL, Michalek SM, Qureshi N, Eaton LJ, Rallabhandi P, Cuesta N, Vogel SN . Immunologic consequences of Francisella tularensis live vaccine strain infection: role of the innate immune response in infection and immunity[J]. J Immunol, 2006,176(11):6888-6899
doi: 10.4049/jimmunol.176.11.6888
[16] Diamond G, Legarda D , Ryan LK.The innate immune response of the respiratory epithelium[J].Immunol Rev, 2000, 173 27-38
doi: 10.1034/j.1600-065X.2000.917304.x pmid: 10719665
[17] Li X, Yang R, Zang Q, Hu YX . Pharmacological actions of glucocorticoids: progress in mechanisms[J]. Guo Ji Yao Xue Yan Jiu Za Zhi, 2009,36(1):27-30
李鑫, 杨蕊, 臧强, 胡永秀 . 糖皮质激素的药理作用机制研究进展[J]. 国际药学研究杂志, 2009,36(1):27-30
[18] Sakaguchi S . Naturally arising Foxp3-expressing CD25 + CD4+ regulatory T cells in immunological tolerance to self and non-self [J]. Nat Immunol, 2005,6(4):345-352
[19] Zhu XP, Yin WY, Jiang HY, Fei Y . The serum levels of immunoglobulin/immunoglobulin G subclass and the level of CD molecules in children with recurrent respiratory tract infection[J]. Lin Chuang Er Ke Za Zhi, 2010,28(2):135-137
朱晓萍, 尹文艳, 蒋红雨, 费樱 . 反复呼吸道感染患儿血清免疫球蛋白、IgG亚类及细胞免疫水平[J]. 临床儿科杂志, 2010,28(2):135-137
[20] Chen X, Murakami T, Oppenheim JJ, Zack Howard OM . Differential response of murine CD4+CD25+ and CD4+CD25-T cells to dexamethasone-induced cell death[J]. Eur J Immunol, 2004,34(3):859-869
doi: 10.1002/eji.v34:3
[1] Victor Abou Nehmi Filho, Gilson Masahiro Murata, Ruan Carlos Macêdo de Moraes, Gabriely Cristina Alves Lima, Danielle Araujo de Miranda, Katrin Radloff, Raquel Galvão Figuerêdo Costa, Joyce de Cassia Rosa de Jesus, Jéssica Alves De Freitas, Nayara Izabel Viana, Ruan Pimenta, Katia Ramos Moreira Leite, José Pinhata Otoch, Ana Flávia Marçal Pessoa. A novel supplement with yeast β-glucan, prebiotic, minerals and Silybum marianum synergistically modulates metabolic and inflammatory pathways and improves steatosis in obese mice. Journal of Integrative Medicine, 2021, 19(5): 439-450.
[2] Hao Gou, Ruo-chen Huang, Yong-hua Su, Wei Li. Design of dual targeting immunomicelles loaded with bufalin and study of their anti-tumor effect on liver cancer. Journal of Integrative Medicine, 2021, 19(5): 408-417.
[3] Yi Ran Hu, San Li Xing, Chuan Chen, Ding Zhu Shen, Jiu Lin Chen. Tiaoxin Recipe, a Chinese herbal formula, inhibits microRNA-34a expression in the APPswe/PS1ΔE9 mouse model of Alzheimer's disease. Journal of Integrative Medicine, 2019, 17(6): 404-409.
[4] Bhesh Raj Sharma, Dong Wook Kim, Dong Young Rhyu. Korean Chungtaejeon tea extract attenuates weight gain in C57BL/6J-Lep ob/ob mice and regulates adipogenesis and lipolysis in 3T3-L1 adipocytes. Journal of Integrative Medicine, 2017, 15(1): 56-63.
[5] Yu-fan Shen, Xue Zhang, Ying Wang, Fan-fan Cao, Georges Uzan, Bin Peng, Deng-hai Zhang. Celastrol targets IRAKs to block Toll-like receptor 4-mediated nuclear factor-κB activation. Journal of Integrative Medicine, 2016, 14(3): 203-208.
[6] Shi-li Jiang, Xu-dong Hu, Ping Liu. Immunomodulation and liver protection of Yinchenhao decoction against concanavalin A-induced chronic liver injury in mice. Journal of Integrative Medicine, 2015, 13(4): 262-268.
[7] Elizabeth Abidemi Balogun, Sylvia Orume Malomo, Joseph Oluwatope Adebayo, Ahmed Adebayo Ishola, Ayodele Olufemi Soladoye, Lawrence Aderemi Olatunji, Olatunji Matthew Kolawole, Stephen Olubunmi Oguntoye, Abiola Samuel Babatunde, Oluwole Busayo Akinola . In vivo antimalarial activity and toxicological effects of methanolic extract of Cocos nucifera (Dwarf red variety) husk fibre. Journal of Integrative Medicine, 2014, 12(6): 504-511.
[8] Md. Asadujjaman, Ahmed Ullah Mishuk, Md. Aslam Hossain, Utpal Kumar Karmakar. Medicinal potential of Passiflora foetida L. plant extracts: Biological and pharmacological activities. Journal of Integrative Medicine, 2014, 12(2): 121-126.
[9] Marjan Nassiri-Asl, Taghi Naserpour Farivar, Esmail Abbasi, Hamid Reza Sadeghnia, Mehdi Sheikhi, Mina Lotfizadeh, Parisa Bazahang. Effects of rutin on oxidative stress in mice with kainic acid-induced seizure. Journal of Chinese Integrative Medicine, 2013, 11(5): 337-342.
[10] Md. Khirul Islam, Imran Mahmud, Sanjib Saha, Asit Baron Sarker, Himangsu Mondal, A.S.M. Monjur-Al-Hossain, Md. Anisuzzman. Preliminary pharmacological evaluation of Alocasia indica Schott tuber. Journal of Chinese Integrative Medicine, 2013, 11(5): 343-351.
[11] Sanjib Saha, Faroque Hossain, Md. Anisuzzman, Md. Khirul Islam. Pharmacological evaluation of Musa seminifera Lour. Fruit. Journal of Chinese Integrative Medicine, 2013, 11(4): 253-261.
[12] Sunil A. Nirmal, Jayashri M. Ingale, Shashikant R. Pattan, Sanjay B. Bhawar. Amaranthus roxburghianus root extract in combination with piperine as a potential treatment of ulcerative colitis in mice. Journal of Chinese Integrative Medicine, 2013, 11(3): 206-212.
[13] Lin Wang, Guan-liang Wang, Jia-han Liu, Di Li, De-zeng Zhu, Liang-neng Wu . Effects of ursolic acid in ameliorating insulin resistance in liver of KKAy mice via peroxisome proliferator-activated receptors α and γ. Journal of Chinese Integrative Medicine, 2012, 10(7): 793-799.
[14] Mei-jiao Mao, Jun-ping Hu, Cong Wang, Yi-yi Zhang, Ping Liu. Effects of Chinese herbal medicine Guanxinkang on expression of PPARγ-LXRα-ABCA1 pathway in ApoE-knockout mice with atherosclerosis. Journal of Chinese Integrative Medicine, 2012, 10(7): 814-820.
[15] Yi-tao Zhao , Xin-guang Zhang , Li Bai , Li-qing Li , Jian-er Yu . Effects of Chinese herbal medicine Pingchuan Formula on airway inflammation, interferon-γ and interleukin-4 in mice with asthma. Journal of Chinese Integrative Medicine, 2012, 10(7): 807-813.
Full text



[1] Dong Yang, Yong-ping Du, Qing Shen, Wei Chen, Yan Yu, Guang-lei Chen. Expression of alpha-smooth muscle actin in renal tubulointerstitium in patients with kidney collateral stasis. Journal of Chinese Integrative Medicine, 2008, 6(1): 41-44
[2] Hai-feng Wei, Bai-liu Ya, Ling Zhao, Cui-fei Ye, Li Zhang, Lin Li. Evaluation of tongue manifestation of blood stasis syndrome and its relationship with blood rheological disorder in a rat model of transient brain ischemia. Journal of Chinese Integrative Medicine, 2008, 6(1): 73-76
[3] Xi Lin, Jian-ping Liu. Herbal medicines for viral myocarditis. Journal of Chinese Integrative Medicine, 2008, 6(1): 76
[4] Xi Lin, Jian-ping Liu. Tai chi for treating rheumatoid arthritis. Journal of Chinese Integrative Medicine, 2008, 6(1): 82
[5] Liang-ping Hu, Hui Gao. Discrimination of errors in statistical analysis of medical papers published in the first issue of 2006 in Journal of Chinese Integrative Medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 98-106
[6] Yan-bo Zhu , Qi Wang, Cheng-yu Wu, Guo-ming Pang, Jian-xiong Zhao, Shi-lin Shen, Zhong-yuan Xia , Xue Yan . Logistic regression analysis on relationships between traditional Chinese medicine constitutional types and overweight or obesity. Journal of Chinese Integrative Medicine, 2010, 8(11): 1023-1035
[7] Wei Xu, Meng Shi, Jian-gang Liu, Cheng-long Wang . Collagen protein expressions in ischemic myocardium of rats with acute myocardial infarction and effects of qi-tonifying, yin-tonifying and blood-activating herbs and detoxifying and blood-activating herbs. Journal of Chinese Integrative Medicine, 2010, 8(11): 1041-1047
[8] Tao Wang , Feng Qin. Effects of Chinese herbal medicine Xiaoyao Powder on monoamine neurotransmitters in hippocampus of rats with postpartum depression. Journal of Chinese Integrative Medicine, 2010, 8(11): 1075-1079
[9] Ying Xu , Chang-chun Zeng , Xiu-yu Cai , Rong-ping Guo , Guang Nie , Ying Jin. Chromaticity and optical spectrum colorimetry of the tongue color in different syndromes of primary hepatic carcinoma. Journal of Chinese Integrative Medicine, 2012, 10(11): 1263-1271
[10] Xiang-ying Mao , Qin Bian , Zi-yin Shen. Analysis of the osteogenetic effects exerted on mesenchymal stem cell strain C3H10T1/2 by icariin via MAPK signaling pathway in vitro. Journal of Chinese Integrative Medicine, 2012, 10(11): 1272-1278