Search JIM Advanced Search

Journal of Integrative Medicine ›› 2014, Vol. 12 ›› Issue (5): 417-424.doi: 10.1016/S2095-4964(14)60040-6

• Research Article • Previous Articles     Next Articles

Electroacupuncture at Feishu (BL13) and Zusanli (ST36) down-regulates the expression of orexins and their receptors in rats with chronic obstructive pulmonary disease

Xin-fang Zhanga, Ji Zhub, Wen-ye Gengc, Shu-jun Zhaoa, Chuan-wei Jianga, Sheng-rong Caia, Miao Chengd, Chuan-yun Zhoud, Zi-bing Liua,d   

  1. Department of Physiology, College of Integrated Chinese and Western Medicine, Anhui University of Chinese Medicine, Hefei 230038, Anhui Province, China
    The Third Clinical College of Zhejiang Chinese Medical University, Hangzhou 310005, Zhejiang Province, China
    Department of Pharmacology, Shanghai Medical College of Fudan University, Shanghai 200032, China
    Institute of Acupuncture and Meridian, College of Acupuncture and Osteology, Anhui University of Chinese Medicine, Hefei 230038, Anhui Province, China
  • Received:2014-01-29 Accepted:2014-05-19 Online:2014-09-10 Published:2014-09-15


Inflammation and lung function decline are the main pathophysiological features of chronic obstructive pulmonary disease (COPD). Acupuncture can improve lung function in patients with COPD, but the underlying mechanisms are not well understood. Orexins (OXs), which are found in peripheral plasma, are neuropeptides that regulate respiration and their levels are related to COPD. Therefore, we hypothesized that acupuncture might alter OXs, reduce lung inflammation and improve lung function in COPD.


COPD was induced in rats by exposure to cigarette smoke for 8 weeks and injecting with lipopolysaccharide twice. Electroacupuncture (EA) was performed at Feishu (BL13) and Zusanli (ST36) for 30 min/d for 2 weeks. Rat lung function and morphology were assessed after EA. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in bronchoalveolar lavage fluid (BALF) and orexin A and B levels in the lung tissue were detected by enzyme-linked immunosorbent assay. OX receptor mRNA levels and immunopositive cells were assessed with real-time polymerase chain reaction and immunohistochemical methods, respectively. The relationships among lung function, cell factors, and OX levels were analyzed by Pearson correlation analyses. 


Compared with the control group, lung function was significantly decreased in the rats with COPD (P<0.05). There were obvious increases in TNF-α and IL-1β levels in BALF (P<0.05 and P<0.01, respectively), orexin A level in lung tissue (P<0.01; but not orexin B) and mRNA expressions of OX 1 type receptor (OXR1) and OX 2 type receptor (OXR2) in lung tissue (P<0.05 and P<0.01, respectively); the integrative optical densities (IODs) of both receptors were greater in the COPD group (P<0.05). For rats with COPD subjected to EA, lung function was improved (P<0.05). There were notable decreases in TNF-α and IL-1β levels (P<0.05 and <0.01, respectively) in BALF. Orexin A, but not orexin B, levels in lung tissue also decreased (P<0.01), as did mRNA expression of OX1R and OX2R in lung tissue (P<0.05 and P<0.01, respectively). Receptor IODs were also reduced after EA treatment (P<0.05). Furthermore, orexin A levels and ratio of forced expiratory volume in 0.3 s to forced vital capacity were strongly negatively correlated (P<0.01), and orexin A was positively correlated with TNF-α and IL-1β (P<0.001 and P<0.05, respectively).


EA at Zusanli and Feishu improved lung function of rats with COPD and had an anti-inflammatory effect, which may be related to down-regulation of OXA and its receptors.

Key words: Pulmonary disease, Chronic obstructive, Electroacupuncture, Point BL13 (Feishu), Point ST36 (Zusanli), Orexin, Anti-inflammation, Rats


Figure 1

Comparison of lung histology in different groupsA: Hematoxylin-eosin staining showed large lung alveoli and thin alveolar septum in the control and EA groups, but small lung alveoli and thickened alveolar septum in the COPD group. B: The histogram showed the total area ratio of lung alveolus to alveolar septum (A/t). Data in Figure 1B are expressed as mean ± standard deviation (n=6). *P<0.05, vs control group. EA: electroacupuncture; COPD: chronic obstructive pulmonary disease."



Figure 2

Orexin receptor expression in rat lung A-F: Immunohistochemical staining showing immunoreactive cells (arrows) for orexin 1 type receptor (OX1R) and orexin 2 type receptor (OX2R) in the lung of the control (A and D), COPD (B and E) and EA group (C and F) rats. Bar, 100 μm. G: Comparison of the integrative optical density (IOD) of OX1R or OX2R. H: Comparison of OX1R and OX2R mRNA levels. Data in Figure 2G and Figure 2H are expressed as mean±standard deviation and analyzed by one-way analysis of variance followed by least-significant difference t tests for comparison of means between two groups (n=6). *P<0.05, vs control group; △P<0.05, vs COPD group. EA: electroacupuncture; COPD: chronic obstructive pulmonary disease."

[1] Fabbri LM, Hurd SS . Global Strategy for the Diagnosis, Management and Prevention of COPD: 2003 update[J]. Eur Respir J, 2003,22(1):1-2
doi: 10.1183/09031936.03.00063703
[2] Corhay JL, Nguyen Dang D, Louis R . Exacerbations in COPD: a burden to curtail[J]. Rev Med Liege, 2006,61(10):691-696
[3] Neumeister W, Kuhlemann H, Bauer T, Krause S, Schultze-Werninghaus G, Rasche K . Effect of acupuncture on quality of life, mouth occlusion pressures and lung function in COPD[J]. Med Klin (Munich), 1999,94(1 Spec No):106-109
[4] Geng WY, Liu ZB, Song NN, Zhang GH, Jin WZ, Li L, Cao YX, Zhu DN, Shen LL . Effects of electroacupuncture at Zusanli(ST36) on inflammatory cytokines in a rat model of smoke-induced chronic obstructive pulmonary disease[J]. J Integr Med, 2013,11(3):213-219
doi: 10.3736/jintegrmed2013024
[5] Lin YY, Wang YP, Lu HY, Guo XC, Liu X, Wu CB, Xu YH . Plasma pharmacokinetics and lung distribution of tetrahydropalmatine after topical application of cold asthma recipe extract: Feishu(BL 13) versus non-Feishu acupoint[J]. J Ethnopharmacol, 2014,153(2):454-460
doi: 10.1016/j.jep.2014.03.009
[6] Li R, Li WJ, Cai YN, Li ZG, Luo Q, Zhou MJ, Li CX, Li FH, Liu MF . Effects of moxibustion at Feishu(BL13) and Gaohuang(BL43) on expression of TGF-β in the bleomycin -induced pulmonary fibrosis[J]. Zhongguo Zhen Jiu, 2005,25(11):790-792
[7] Yu JB, Dong SA, Luo XQ, Gong LR, Zhang Y, Wang M, Cao XS, Liu DQ . Role of HO-1 in protective effect of electro-acupuncture against endotoxin shock-induced acute lung injury in rabbits[J]. Exp Biol Med (Maywood), 2013,238(6):705-712
doi: 10.1177/1535370213489487
[8] Sakurai T, Amemiya A, Ishii M, Matsuzaki I, Chemelli RM, Tanaka H, Williams SC, Richardson JA, Kozlowski GP, Wilson S, Arch JR, Buckingham RE, Haynes AC, Carr SA, Annan RS , McNulty DE, Liu WS, Terrett JA, Elshourbagy NA, Bergsma DJ, Yanagisawa M.Orexins and orexin receptors: a family of hypothalamic neuropeptides and G protein-coupled receptors that regulate feeding behavior[J]. Cell, 1998,92(4):573-585
doi: 10.1016/S0092-8674(00)80949-6
[9] Young JK, Wu M, Manaye KF, Kc P, Allard JS, Mack SO, Haxhiu MA . Orexin stimulates breathing via medullary and spinal pathways[J]. J Appl Physiol (1985), 2005,98(4):1387-1395
doi: 10.1152/japplphysiol.00914.2004
[10] Zhang W, Fukuda Y, Kuwaki T . Respiratory and cardiovascular actions of orexin-A in mice[J]. Neurosci Lett, 2005,385(2):131-136
doi: 10.1016/j.neulet.2005.05.032
[11] Liu ZB, Song NN, Geng WY, Jin WZ, Li L, Cao YX, Qian Y, Zhu DN, Shen LL . Orexin-A and respiration in a rat model of smoke-induced chronic obstructive pulmonary disease[J]. Clin Exp Pharmacol Physiol, 2010,37(10):963-968
doi: 10.1111/j.1440-1681.2010.05411.x
[12] Kastin AJ, Akerstrom V . Orexin A but not orexin B rapidly enters brain from blood by simple diffusion[J]. J Pharmacol Exp Ther, 1999,289(1):219-223
[13] Matsumura T, Nakayama M, Satoh H, Naito A, Kamahara K, Sekizawa K . Plasma orexin-A levels and body composition in COPD[J]. Chest, 2003,123(4):1060-1065
doi: 10.1378/chest.123.4.1060
[14] Nie YC, Wu H, Li PB, Luo YL, Zhang CC, Shen JG, Su WW . Characteristic comparison of three rat models induced by cigarette smoke or combined with LPS: to establish a suitable model for study of airway mucus hypersecretion in chronic obstructive pulmonary disease[J]. Pulm Pharmacol Ther, 2012,25(5):349-356
doi: 10.1016/j.pupt.2012.06.004
[15] Liu Z, Liu J, Zhao Y, Cai Y, He L, Xu H, Zhou X, Yan S, Lao L, Liu B . The efficacy and safety study of electro-acupuncture for severe chronic functional constipation: study protocol for a multicenter, randomized, controlled trial[J]. Trials, 2013,15(14):176
[16] Livak KJ, Schmittgen TD . Analysis of relative gene expression data using real-time quantitative PCR and the 2- ΔΔ CT method [J]. Methods, 2001,25(4):402-408
doi: 10.1006/meth.2001.1262
[17] Martin JG, Tamaoka M . Rat models of asthma and chronic obstructive lung disease[J]. Pulm Pharmacol Ther, 2006,19(6):377-385
doi: 10.1016/j.pupt.2005.10.005
[18] Tian L, Huang YX, Tian M, Gao W, Chang Q . Downregulation of electroacupuncture at ST36 on TNF-alpha in rats with ulcerative colitis[J]. World J Gastroenterol, 2003,9(5):1028-1033
doi: 10.3748/wjg.v9.i5.1028
[19] Yao WP, Wang S, Han L, Ma JQ, Shen Y . Effects of different frequencies of electro-acupuncture at shuigou(GV 26) on recovery of motor function in rats with focal cerebral ischemic injury[J]. J Tradit Chin Med, 2012,32(1):99-104
doi: 10.1016/S0254-6272(12)60040-5
[20] Arihara Z, Takahashi K, Murakami O, Totsune K, Sone M, Satoh F, Ito S, Mouri T . Immunoreactive orexin-A in human plasma[J]. Peptides, 2001,22(1):139-142
doi: 10.1016/S0196-9781(00)00369-7
[21] Jhren O, Neidert SJ, Kummer M, Dendorfer A, Dominiak P . Prepro-orexin and orexin receptor mRNAs are differentially expressed in peripheral tissues of male and female rats[J]. Endocrinology, 2001,142(8):3324-3331
doi: 10.1210/endo.142.8.8299
[1] Xin-fang Zhang, Shui-ying Xiang, Wen-ye Geng, Wen-juan Cong, Jing Lu, Chuan-wei Jiang, Kun Wang, Zi-bing Liu . Electro-acupuncture regulates the cholinergic anti-inflammatory pathway in a rat model of chronic obstructive pulmonary disease. Journal of Integrative Medicine, 2018, 16(6): 418-426.
[2] Morufu Eyitayo Balogun, Elizabeth Enohnyaket Besong, Jacinta Nkechi Obimma, Ogochukwu Sophia Mbamalu, Fankou Serges Athanase Djobissie. Protective roles of Vigna subterranea (Bambara nut) in rats with aspirin-induced gastric mucosal injury. Journal of Integrative Medicine, 2018, 16(5): 342-349.
[3] Lucky Legbosi Nwidu, Raphael Ellis Teme. Hot aqueous leaf extract of Lasianthera africana (Icacinaceae) attenuates rifampicin-isoniazid-induced hepatotoxicity. Journal of Integrative Medicine, 2018, 16(4): 263-272.
[4] Kylie Connolly, Douglas Jackson, Candice Pullen, Andrew Fenning. Alpha-adrenoceptor antagonism by Crassostrea gigas oyster extract inhibits noradrenaline-induced vascular contraction in Wistar rats. Journal of Integrative Medicine, 2015, 13(3): 194-200.
[5] Hussein O. B. Oloyede, Matthew C. Adaja, Taofeek O. Ajiboye, Musa O. Salawu. Anti-ulcerogenic activity of aqueous extract of Carica papaya seed on indomethacin-induced peptic ulcer in male albino rats. Journal of Integrative Medicine, 2015, 13(2): 105-114.
[6] Udhaya Lavinya Baskaran, Sherry Joseph Martin, Rasool Mahaboobkhan, Sabina Evan Prince. Protective role of Triphala, an Indian traditional herbal formulation, against the nephrotoxic effects of bromobenzene in Wistar albino rats. Journal of Integrative Medicine, 2015, 13(2): 115-121.
[7] Anirudha A. Lande, Shirishkumar D. Ambavade, Uma S. Swami, Prafulla P. Adkara Prashant D. Ambavade, Arun B. Waghamare. Saponins isolated from roots of Chlorophytum borivilianum reduce acute and chronic inflammation and histone deacetylase. Journal of Integrative Medicine, 2015, 13(1): 25-33.
[8] 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.
[9] Meng-yue Liu, Cheng-wei Wang, Zhou-peng Wu, Ning Li. Electroacupuncture for the prevention of postoperative gastrointestinal dysfunction in patients undergoing vascular surgery under general anesthesia: Study protocol for a prospective practical randomized controlled trial. Journal of Integrative Medicine, 2014, 12(6): 512-519.
[10] Urmila Aswar, Mayuri Gurav, Ganesh More, Khaled Rashed, Manoj Aswar. Effect of aqueous extract of Solanum xanthocarpum Schrad. & Wendl. on postmenopausal syndrome in ovariectomized rats. Journal of Integrative Medicine, 2014, 12(5): 439-446.
[11] Benjamin Perry, Junzeng Zhang, Tarek Saleh, Yanwen Wang. Liuwei Dihuang, a traditional Chinese herbal formula, suppresses chronic inflammation and oxidative stress in obese rats. Journal of Integrative Medicine, 2014, 12(5): 447-454.
[12] Upendarrao Golla, Praveen Kumar Gajam, Solomon Sunder Bhimathati. Evaluation of diuretic and laxative activity of hydro-alcoholic extract of Desmostachya bipinnata (L.) Stapf in rats. Journal of Integrative Medicine, 2014, 12(4): 372-378.
[13] Qian Zhang, Xin-hua Xiao, Ming Li, Wen-hui Li, Miao Yu, Hua-bing Zhang, Fan Ping, Zhi-xin Wang, Jia Zheng ​. Chromium-containing traditional Chinese medicine, Tianmai Xiaoke Tablet improves blood glucose through activating insulin-signaling pathway and inhibiting PTP1B and PCK2 in diabetic rats. Journal of Integrative Medicine, 2014, 12(3): 162-170.
[14] Amir Rashidian​, Saeed Mehrzadi, Ali Reza Ghannadi, Parvin Mahzooni, Samira Sadr, Mohsen Minaiyan​. Protective effect of ginger volatile oil against acetic acid-induced colitis in rats: A light microscopic evaluation. Journal of Integrative Medicine, 2014, 12(2): 115-120.
[15] Dilpesh Jain​, Manish Kumar Bansal, Rahul Dalvi​, Aman Upganlawar​, Rahul Somani​. Protective effect of diosmin against diabetic neuropathy in experimental rats. Journal of Integrative Medicine, 2014, 12(1): 35-41.
Full text



[1] Yi-ting He, Qing-lin Zha, Jian-ping Yu, Yong Tan, Cheng Lu, Ai-ping Lv. Principal factor analysis of symptoms of rheumatoid arthritis and their correlations with efficacy of traditional Chinese medicine and Western medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 32-36
[2] 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
[3] A-gao Zhou, Yong Zhang, Gang Kui, De-Yun Kong, Hai-liang Ge, Qiu-hua Ren, Jia-rong Dong, Sheng Hong, Xu-ming Mao, Yin Wang, Hui-zheng Zhang, Shu-jun Wang. Influence of traditional Chinese compound recipes with different efficacy on body weight, tumor weight and immune function in H22 cancer-bearing mice. Journal of Chinese Integrative Medicine, 2008, 6(1): 77-82
[4] Xi Lin, Jian-ping Liu. Tai chi for treating rheumatoid arthritis. Journal of Chinese Integrative Medicine, 2008, 6(1): 82
[5] Guo-hong Yuan, Xiao-jing Pang, He-chao Ma. Synergic effects of Danggui Buxue Decoction in reducing toxicity of cytoxan in tumor-bearing mice. Journal of Chinese Integrative Medicine, 2008, 6(1): 83-88
[6] Li Zhou, Hong-xing Zhang, Ling-guang Liu, Wen-jun Wan. Effect of electro-acupuncture at Fenglong (GV 16) on nitric oxide and endothelin in rats with hyperlipidemia. Journal of Chinese Integrative Medicine, 2008, 6(1): 89-92
[7] Ning-qun Wang, Liang-duo Jiang, Zong-xing Li. Research progress in asthma-related quality of life. Journal of Chinese Integrative Medicine, 2008, 6(1): 93-97
[8] Jing-yuan Mao, Chang-xiao Liu, Heng-he Wang, Guang-li Wei , Zhen-peng Zhang, Jie Xing, Wang Xian liang , Ying-fei Bi . Effects of Shenmai Injection on serum concentration and pharmacokinetics of digoxin in dogs with heart failure. Journal of Chinese Integrative Medicine, 2010, 8(11): 1070-1074
[9] Liang-ping Hu , Xiao-lei Bao, Xue Guan. Statistical inference and experimental design of univariate qualitative data of single-group design. Journal of Chinese Integrative Medicine, 2010, 8(11): 1085-1089
[10] Zhao-guo Li . A discussion of English translation of 1995 and 1997 Chinese National Standards of Traditional Chinese Medical Terminologies for Clinical Diagnosis and Treatment. Journal of Chinese Integrative Medicine, 2010, 8(11): 1090-1096