Buyang Huanwu Decoction (补阳还五汤) Attenuates Glial Scar by Downregulating the Expression of Leukemia Inhibitory Factor in Intracerebral Hemorrhagic Rats

  • Xiao Kang
  • Hua-jun ZhouEmail author
  • Jian Yang
  • Jian-hua Zhong
  • Tao Tang
  • Han-jin Cui Cui
  • Jing-hua Zhou
  • Zhi-gang Mei
Original Article



To evaluate the effect of Buyang Huanwu Decoction (补阳还五汤, BYHWD) on glial scar after intracerebral hemorrhage (ICH) and investigate the underlying mechanism.


Collagenase type VII (0.5 U) was injected stereotaxically into right globus pallidus to induce ICH model. One hundred and twenty Sprague-Dawley rats were randomly divided into 3 groups according to a random number table, including normal group (n=40), ICH model group (n=40) and BYHWD group (n=40), respectively. After ICH, the rats in the BYHWD group were intragastrically administered with BYHWD (4.36 g/kg) once a day for 21 days, while the rats in ICH group were administered with equal volume of distilled water for 21 days, respectively. Double immunolabeling was performed for proliferating cell nuclear antigen (PCNA)+/glial fifibrillary acidic protein (GFAP)+ nuclei. The expression of GFAP and leukemia inhibitory factor (LIF) was evaluated by immunohistochemistry and quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR).


The astrocytes with hypertrophied morphology around the hematoma was observed on day 3 after ICH. The number of GFAP positive cells and GFAP mRNA levels increased notably on day 3 and reached the peak on day 14 post-ICH (P<0.01). PCNA+/GFAP+ nuclei were observed around the hematoma and reached the peak on day 14 post-ICH (P<0.01). In addition, LIF-positive astrocytes and LIF mRNA level in the hemorrhagic region increased signifificantly till day 14 post-ICH (P<0.01). However, BYHWD not only reduced the number of PCNA+/GFAP+ nuclei, but also decreased GFAP and LIF levels (P<0.05).


BYHWD could attenuate ICH-induced glial scar by downregulating the expression of LIF in the rats.


Buyang Huanwu Decoction intracerebral hemorrhage glial scar leukemia inhibitory factor Chinese medicine 


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  1. 1.
    Wang W, Jiang B, Sun H, Ru X, Sun D, Wang L, et al. Prevalence, incidence and mortality of stroke in China: results from a nationwide population-based survey of 480,687 adults. Circulation 2017;135:759–771.CrossRefGoogle Scholar
  2. 2.
    Herrington WG. Statin use and outcome after intracerebral hemorrhage: case-control study and meta-analysis. Neurology 2011;77:2073.CrossRefGoogle Scholar
  3. 3.
    Dossi E, Vasile F, Rouach N. Human astrocytes in the diseased brain. Brain Res Bull 2018;136:139–156.CrossRefGoogle Scholar
  4. 4.
    Sukumari-Ramesh S, Alleyne CH Jr, Dhandapani KM. Astrocyte-specific expression of survivin after intracerebral hemorrhage in mice: a possible role in reactive gliosis? J Neurotrauma 2012;29:2798–2804.CrossRefGoogle Scholar
  5. 5.
    Pekny M, Wilhelmsson U, Pekna M. The dual role of astrocyte activation and reactive gliosis. Neurosci Lett 2014;565:30–38.CrossRefGoogle Scholar
  6. 6.
    Onishi K, Zandstra PW. LIF signaling in stem cells and development. Development 2015;142:2230–2236.CrossRefGoogle Scholar
  7. 7.
    Nakanishi M, Niidome T, Matsuda S, Akaike A, Kihara T, Sugimoto H. Microglia-derived interleukin-6 and leukaemia inhibitory factor promote astrocytic differentiation of neural stem/progenitor cells. Eur J Neurosci 2007;25:649–658.CrossRefGoogle Scholar
  8. 8.
    Cohen JE, Fields RD. Activity-dependent neuron-glial signaling by ATP and leukemia-inhibitory factor promotes hippocampal glial cell development. Neuron Glia Biol 2008;4:43–55.CrossRefGoogle Scholar
  9. 9.
    Sugiura S, Lahav R, Han J, Kou SY, Banner LR, de Pablo F, et al. Leukaemia inhibitory factor is required for normal inflammatory responses to injury in the peripheral and central nervous systems in vivo and is chemotactic for macrophages in vitro. Eur J Neurosci 2000;12:457–466.CrossRefGoogle Scholar
  10. 10.
    Zhou HJ, Yang X, Cui HJ, Tang T, Zhong JH, Luo JK, et al. Leukemia inhibitory factor contributes to reactive astrogliosis via activation of signal transducer and activator of transcription 3 signaling after intracerebral hemorrhage in rats. J Neurotrauma 2017;34:1658–1665.CrossRefGoogle Scholar
  11. 11.
    Wei RL, Teng HJ, Yin B, Xu Y, Du Y, He FP, et al. A systematic review and meta-analysis of Buyang Huanwu Decoction in animal model of focal cerebral ischemia. Evid Based Complement Alternat Med 2013;2013:138484.Google Scholar
  12. 12.
    Liu BY, Song XL, Yi J, Chen XM, Yu Y, Liu H, et al. Buyang Huanwu Decoction reduces infarct volume and enhances estradiol and estradiol receptor concentration in ovariectomized rats after middle cerebral artery occlusion. Chin J Integr Med 2014;20:782–786.CrossRefGoogle Scholar
  13. 13.
    Cai G, Liu B, Liu W, Tan X, Rong J, Chen X, et al. Buyang Huanwu Decoction can improve recovery of neurological function, reduce infarction volume, stimulate neural proliferation and modulate VEGF and Flk1 expressions in transient focal cerebral ischaemic rat brains. J Ethnopharmacol 2007;113:292–299.CrossRefGoogle Scholar
  14. 14.
    Li HQ, Wei JJ, Xia W, Li JH, Liu AJ, Yin SB, et al. Promoting blood circulation for removing blood stasis therapy for acute intracerebral hemorrhage: a systematic review and metaanalysis. Acta Pharmacol Sin 2015;36:659–675.CrossRefGoogle Scholar
  15. 15.
    Yang AL, Liang QH, Cui HJ, Zhou HJ, Luo JK, Tang T. Angiogenesis opens a way for Chinese medicine to treat stroke. Chin J Integr Med 2013;19:815–819.CrossRefGoogle Scholar
  16. 16.
    Rosenberg GA, Mun-Bryce S, Wesley M, Kornfeld M. Collagenase-induced intracerebral hemorrhage in rats. Stroke 1990;21:801–807.CrossRefGoogle Scholar
  17. 17.
    Suzuki S, Tanaka K, Nogawa S, Ito D, Dembo T, Kosakai A, et al. Immunohistochemical detection of leukemia inhibitory factor after focal cerebral ischemia in rats. J Cereb Blood Flow Metab 2000;20:661–668.CrossRefGoogle Scholar
  18. 18.
    Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method. Methods 2001;25:402–408.CrossRefGoogle Scholar
  19. 19.
    Chen ZZ, Yang DD, Zhao Z, Yan H, Ji J, Sun XL. Memantine mediates neuroprotection via regulating neurovascular unit in a mouse model of focal cerebral ischemia. Life Sci 2016;150:8–14.CrossRefGoogle Scholar
  20. 20.
    Zhang L, Zhang ZG, Chopp M. The neurovascular unit and combination treatment strategies for stroke. Trends Pharmacol Sci 2012;33:415–422.CrossRefGoogle Scholar
  21. 21.
    Liu B, Cai G, Yi J, Chen X. Buyang Huanwu Decoction regulates neural stem cell behavior in ischemic brain. Neural Regen Res 2013;8:2336–2342.Google Scholar
  22. 22.
    Zhang M, Chai Y, Liu T, Xu N, Yang C. Synergistic effects of Buyang Huanwu Decoction and embryonic neural stem cell transplantation on the recovery of neurological function in a rat model of spinal cord injury. Exp Ther Med 2015;9:1141–1148.CrossRefGoogle Scholar
  23. 23.
    Munakata M, Shirakawa H, Nagayasu K, Miyanohara J, Miyake T, Nakagawa T, et al. Transient receptor potential canonical 3 inhibitor Pyr3 improves outcomes and attenuates astrogliosis after intracerebral hemorrhage in mice. Stroke 2013;44:1981–1987.CrossRefGoogle Scholar
  24. 24.
    Zhao YY, Yuan Y, Chen Y, Jiang L, Liao RJ, Wang L, et al. Histamine promotes locomotion recovery after spinal cord hemisection via inhibiting astrocytic scar formation. CNS Neurosci Ther 2015;21:454–462.CrossRefGoogle Scholar
  25. 25.
    Kerr BJ, Patterson PH. Potent pro-inflammatory actions of leukemia inhibitory factor in the spinal cord of the adult mouse. Exp Neurol 2004;188:391–407.CrossRefGoogle Scholar
  26. 26.
    Zhong JH, Zhou HJ, Tang T, Cui HJ, Yang AL, Zhang QM, et al. Activation of the Notch-1 signaling pathway may be involved in intracerebral hemorrhage-induced reactive astrogliosis in rats. J Neurosurg 2018;129:732–739.CrossRefGoogle Scholar
  27. 27.
    Ding W, Yu P, Liu W, Zhou L, Guan LI, Lin R. Buyang Huanwu Decoction increases the expression of glutamate transporter-1 and glutamate synthetase in association with PACAP-38 following focal ischemia. Biomed Rep 2015;3:651–656.CrossRefGoogle Scholar
  28. 28.
    Fitch MT, Silver J. CNS injury, glial scars, and inflammation: inhibitory extracellular matrices and regeneration failure. Exp Neurol 2008;209:294–301.CrossRefGoogle Scholar
  29. 29.
    Liu FT, Xu SM, Xiang ZH, Li XN, Li J, Yuan HB, et al. Molecular hydrogen suppresses reactive astrogliosis related to oxidative injury during spinal cord injury in rats. CNS Neurosci Ther 2014;20:778–786.CrossRefGoogle Scholar
  30. 30.
    Wang J, Dore S. Inflammation after intracerebral hemorrhage. J Cereb Blood Flow Metab 2007;27:894–908.CrossRefGoogle Scholar
  31. 31.
    Jin Y, Dong L, Wu C, Qin J, Li S, Wang C, et al. Buyang Huanwu Decoction fraction protects against cerebral ischemia/reperfusion injury by attenuating the inflammatory response and cellular apoptosis. Neural Regen Res 2013;8:197–207.Google Scholar
  32. 32.
    Shen J, Zhu Y, Huang K, Jiang H, Shi C, Xiong X, et al. Buyang Huanwu Decoction attenuates H2O2-induced apoptosis by inhibiting reactive oxygen species-mediated mitochondrial dysfunction pathway in human umbilical vein endothelial cells. BMC Complement Altern Med 2016;16:154.CrossRefGoogle Scholar
  33. 33.
    Wang Y, Ren T, Zheng L, Chen H, Ko JK, Auyeung KK. Astragalus saponins inhibits lipopolysaccharide-induced inflammation in mouse macrophages. Am J Chin Med 2016;44:579–593.CrossRefGoogle Scholar
  34. 34.
    Xu J, E XQ, Liu HY, Tian J, Yan JL. Angelica Sinensis attenuates inflammatory reaction in experimental rat models having spinal cord injury. Int J Clin Exp Pathol 2015;8:6779–6785.Google Scholar
  35. 35.
    Di Cesare Mannelli L, Pacini A, Micheli L, Femia AP, Maresca M, Zanardelli M, et al. Astragali Radix: could it be an adjuvant for oxaliplatin-induced neuropathy? Sci Rep 2017;7:42021.CrossRefGoogle Scholar

Copyright information

© Chinese Association of the Integration of Traditional and Western Medicine 2018

Authors and Affiliations

  • Xiao Kang
    • 1
    • 2
  • Hua-jun Zhou
    • 1
    • 2
    • 3
    Email author
  • Jian Yang
    • 3
  • Jian-hua Zhong
    • 4
  • Tao Tang
    • 5
  • Han-jin Cui Cui
    • 5
  • Jing-hua Zhou
    • 1
    • 2
  • Zhi-gang Mei
    • 6
  1. 1.Institute of NeurologyChina Three Gorges UniversityYichang, Hubei ProvinceChina
  2. 2.Department of Neurology, The First College of Clinical Medical SciencesChina Three Gorges UniversityYichang, Hubei ProvinceChina
  3. 3.Key Laboratory of Cardiovascular and Cerebrovascular Diseases Translational MedicineChina Three Gorges UniversityYichang, Hubei ProvinceChina
  4. 4.Department of Intensive Care Unit, The First College of Clinical Medical SciencesChina Three Gorges UniversityYichang, Hubei ProvinceChina
  5. 5.Institute of Integrative Medicine, Xiangya HospitalCentral South UniversityChangshaChina
  6. 6.Medical College of China Three Gorges UniversityYichang, Hubei ProvinceChina

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