Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (8): 763-768.doi: 10.3736/jcim20090811

• Original Experimental Research • Previous Articles     Next Articles

Effects of Chinese medicine for regulating “sea of blood in brain” combined with bone marrow stromal stem cell transplantation on angiogenesis in ischemic brain tissue of rats

Jian-wen Guo,Jun-ya Li, Yan Huang   

  1. Center for Encephalopathy, the Second Hospital, Guangzhou University of Chinese Medicine, Guangzhou 510120, Guangdong Province» China
  • Received:2009-03-18 Accepted:2009-05-26 Online:2009-08-20 Published:2009-08-15
  • Contact: Jian-wen Guo E-mail:drguo@yahoo.cn

Objective

To observe whether Naomai Yihao (NM) Capsule, a compound traditional Chinese herbal medicine for regulating the “sea of blood in brain”, and bone marrow stromal stem cell (BMSC) transplantation could improve angiogenesis in focal cerebral ischemia in rats.

Methods

A rat model of middle cerebral artery occlusion (MCAO) was established. The rats were divided into untreated group, NM group, BMSC group and combination of NM and BMSC group (combined treatment group). Another 8 normal rats were selected as sham-operated group. After 3-, 7- and 14-day reperfusion, behavioral rating scale (BRS) of the rats and histopathology of the cerebral tissue were evaluated. Expression of CD31 in the brain tissue was also measured by immunohistochemical method.
Results

Compared with the untreated group, BRSs of the NM group, BMSC group and combined treatment group were decreased significantly (P=0.000), but there was no interacting effect between NM and BMSC transplantation. Compared with the untreated group, the numbers of CD31 positive cells in NM group, BMSC group and combined treatment group were increased significantly (P=0.000), and there were interacting effects among NM, BMSC transplantation, and the observation time (P<0.01). After 14-day reperfusion, combination of NM and BMSC transplantation could largely increase the number of CD31 positive cells.
Conclusion

NM is able to promote the angiogenesis and neurological impairment improvement in focal cerebral ischemia of rats which are administered with BMSC transplantation, and the effect is reinforced with the extension of treatment time.

Key words: Cerebral ischemia, Bone marrow stromal stem cell, Angiogenesis, Pharmacotherapy, TCD, Rats

Table 1

BRS and CD31 expression of rats in different groups ($\bar{x}$±s)"

YZ NM Time (d) BRS (n=8) CD31 expression (n=6)
No No 3 8.50±0.93 10.27±0.90
7 6.00±1.60 12.90±1.47
14 4.88±0.83 13.67±2.25
Yes No 3 7.87±0.83 14.73±1.33
7 5.13±1.13 21.90±1.71
14 3.63±0.92 25.67±2.05
No Yes 3 6.75±1.04 19.03±2.32
7 4.00±1.31 24.37±2.32
14 2.75±0.71 27.60±1.81
Yes Yes 3 6.13±1.13 17.50±2.72
7 3.00±0.76 25.67±2.72
14 1.63±0.91 31.87±4.11

Figure 1

BRS of rats under the interaction among BMSC transplantation, Naomai Yihao Capsule and treatment duration (Multivariate ANOVA, n=8)"

Table 2

BRS of rats after BMSC transplantation, Naomai Yihao Capsule treatment and combined treatment (Multivariate ANOVA, n=8)"

Source Type Ⅲ sum of squares df Mean square F P Value
Corrected Model 393.646 9 43.738 41.650 0.000
Intercept 2 420.042 1 2 420.042 2 304.483 0.000
NM 92.042 1 92.042 87.647 0.000
YZ 20.167 1 20.167 19.204 0.000
Time 279.646 2 139.823 133.146 0.000
NM*YZ 0.000 1 0.000 0.000 1.000
NM*Time 0.521 2 0.260 0.248 0.781
YZ*Time 1.271 2 0.635 0.605 0.548
Error 90.313 86 1.050
Total 2 904.000 96
Corrected Total 483.958 95

Table 3

Mean and 95% confidence interval of BRS of rats in interaction analysis"

Group NM YZ n Mean SE 95% CI
Untreated N N 8 6.458 0.212 [6.038, 6.879]
YZ N Y 8 5.542 0.212 [5.121, 5.962]
NM Y N 8 4.500 0.212 [4.079, 4.921]
NM+YZ Y Y 8 3.583 0.212 [3.163, 4.004]

Figure 2

Pathological changes of cerebral tissues observed by HE staining (Light microscopy, ×20) A: Sham-operated group; B: Untreated group, after 3-day reperfusion; C: BMSC group, after 3-day reperfusion; D: NM group, after 3-day reperfusion; E: Combined treatment group, after 14-day reperfusion."

Figure 3

Expression of CD31 in different groups detected by immunohistochemical staining (Light microscopy, ×40) A: Sham-operated group; B: Untreated group, after 14-day reperfusion; C: BMSC group, after 7-day reperfusion; D: NM group, after 7-day reperfusion; E: Combined treatment group, after 14-day reperfusion."

Table 4

Number of CD31 positive cells of rats after BMSC transplantation, Naomai Yihao Capsule treatment and combined treatment (Multivariate ANOVA, n=6)"

Source Type Ⅲ sum of squares df Mean square F P value
Corrected Model 3 026.840 9 336.316 65.053 0.000
Intercept 30 079.913 1 3 0079.913 5 818.353 0.000
NM 1 104.892 1 1 104.892 213.719 0.000
YZ 431.935 1 431.935 83.549 0.000
Time 1 068.943 2 534.472 103.383 0.000
NM*YZ 232.022 1 232.022 44.880 0.000
NM*Time 57.304 2 28.652 5.542 0.006
YZ*Time 131.744 2 65.872 12.742 0.000
Error 320.530 62 5.170
Total 3 3427.283 72
Corrected Total 3 347.370 71

Figure 4

Number of CD31 positive cells of rats under the interaction among BMSC transplantation, Naomai Yihao Capsule and treatment duration (Multivariate ANOVA, n=6)"

Table 5

Mean and 95% confidence interval of the number of CD31 positive cells of rats in interaction analysis"

Group NM YZ n Mean SE 95% CI
Untreated N N 6 12.278 0.542 [11.193, 13.362]
YZ N Y 6 20.767 0.542 [19.682, 21.851]
NM Y N 6 23.703 0.542 [22.618, 24.787]
NM+YZ Y Y 6 25.011 0.542 [23.927, 26.095]
[1] 1Department of Disease Control , Ministry of Health of the People’s Republic of China. Major situation of the third survey of national death causes recommended by Ministry of Health [2008-04-29]. .
卫生部疾病预防控制局. 卫生部介绍第三次全国死因调查主要情况 [2008-04-29]. .
[2] Haas S, Weidner N, Winkler J . Adult stem cell therapy in stroke[J]. Curr Opin Neurol, 2005,18(1):59-64
doi: 10.1097/00019052-200502000-00012 pmid: 15655404
[3] Amano H, Hackett NR, Rafii S, Crvstal RG . Thrombopoietin gene transfer-mediated enhancement of angiogenic responses to acute ischemia[J]. Circ Rec, 2005,97(4):337-345
doi: 10.1161/01.RES.0000179534.17668.f8
[4] Chen J, Zhang ZG, Li Y, Wang L, Xu YX, Gautam SC, Lu M, Zhu Z, Chopp M . Intravenous administration of human bone marrow stromal cells induces angiogenesis in the ischemic boundary zone after stroke in rats[J]. Circ Res, 2003,92(6):692-699
doi: 10.1161/01.RES.0000063425.51108.8D
[5] Cai YF, Fu Y, Guo JW, You JS, Wang LX, Liang WX, Huang Y, Huang PX . Randomized multicenter control clinical study on acute ischemic stroke treatment with traditional Chinese medicine[J]. Zhong Yao Cai, 2007,30(9):1192-1195
蔡业峰, 付于, 郭建文, 尤劲松, 王立新, 梁伟雄, 黄燕, 黄培新 . 中医药治疗急性缺血中风的多中心随机对照研究[J]. 中药材, 2007,30(9):1192-1195
[6] Longa EZ, Weinstein PR, Carlson S, Cummins R . Reversible middle cerebral artery occlusion without craniectomy in rats[J]. Stroke, 1989,20(1):84-91
doi: 10.1161/01.STR.20.1.84
[7] Rosenberq GA, Mun-Byree S, Wesley M, Kornfeld M . Collagenase-induced intracerebral hemorrhage in rats[J]. Stroke, 1990,21(5):801-807
doi: 10.1161/01.STR.21.5.801 pmid: 2160142
[8] Hurtado O, Pradillo JM, Alonso-Escolano D, Lorenzo P, Sobrino T, Castillo J, Lizasoain I, Moro MA . Neurorepair versus neuroprotection in stroke[J]. Cerebrovasc Dis, 2006,21(Suppl 2):54-63
doi: 10.1159/000091704 pmid: 16651815
[9] Huang Y, Pei J, Wang X . Outcome and safety assessment of Yin pattern of acute ischemic stroke treated with an herbal medicine protocol[J]. Cerebrovasc Dis, 2007,24(6):483-491
doi: 10.1159/000109749
[10] Newman PJ, Berndt MC, Gorski J , White GC 2nd, Lyman S, Paddock C, Muller WA .PECAM-1(CD31) cloning and relation to adhesion molecules of the immunoglobulin gene superfamily[J]. Science, 1990,247(4947):1219-1222
doi: 10.1126/science.1690453 pmid: 201690453
[11] Voermans C, Rood PM, Hordijk PL , Gerritsen WR, van der Schoot CE.Adhesion molecules involved in transendothelial migration of human hematopoietic progenitor cells[J]. Stem Cells, 2000,18(6):435-443
doi: 10.1634/stemcells.18-6-435
[12] Agu O, Hamilton G, Baker DM, Dashwood MR . Endothelin receptors in the aetiology and pathophysiology of varicose veins[J]. Eur J Vasc Endovasc Surg, 2002,23(2):165-171
doi: 10.1053/ejvs.2001.1569 pmid: 11863336
[1] 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.
[2] 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.
[3] Bing He, Ge Zhang, Ai-ping Lu. Integrative network analysis: Bridging the gap between Western medicine and traditional Chinese medicine. Journal of Integrative Medicine, 2015, 13(3): 133-135.
[4] Oluwatosin Adaramoye, Bettina Erguen, Olubukola Oyebode, Bianca Nitzsche, Michael Höpfner, Klaus Jung, Anja Rabien. Antioxidant, antiangiogenic and antiproliferative activities of root methanol extract of Calliandra portoricensis in human prostate cancer cells. Journal of Integrative Medicine, 2015, 13(3): 185-193.
[5] 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.
[6] 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.
[7] 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.
[8] 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.
[9] 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.
[10] Xin-fang Zhang, Ji Zhu, Wen-ye Geng, Shu-jun Zhao, Chuan-wei Jiang, Sheng-rong Cai, Miao Cheng, Chuan-yun Zhou, Zi-bing Liu. Electroacupuncture at Feishu (BL13) and Zusanli (ST36) down-regulates the expression of orexins and their receptors in rats with chronic obstructive pulmonary disease. Journal of Integrative Medicine, 2014, 12(5): 417-424.
[11] 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.
[12] 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.
[13] 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.
[14] 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.
[15] 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.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
[1] Wei-xiong Liang. Problems-solving strategies in clinical treatment guideline for traditional Chinese medicine and integrative medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 1-4
[2] Jun Cai, Hua Wang, Sheng Zhou, Bin Wu, Hua-rong Song, Zheng-rong Xuan. Effect of Sijunzi Decoction and enteral nutrition on T-cell subsets and nutritional status in patients with gastric cancer after operation: A randomized controlled trial. Journal of Chinese Integrative Medicine, 2008, 6(1): 37-40
[3] 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
[4] Xue-mei Liu, Qi-fu Huang, Yun-ling Zhang, Jin-li Lou, Hong-sheng Liu, Hong Zheng. Effects of Tribulus terrestris L. saponion on apoptosis of cortical neurons induced by hypoxia-reoxygenation in rats. Journal of Chinese Integrative Medicine, 2008, 6(1): 45-50
[5] 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
[6] 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
[7] 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
[8] Jin-zhou Tian, Jing Shi, Xin-qing Zhang, Qi Bi, Xin Ma, Zhi-liang Wang, Xiao-bin Li, Shu-li Shen, Lin Li, Zhen-yun Wu, Li-yan Fang, Xiao-dong Zhao, Ying-chun Miao, Peng-wen Wang, Ying Ren, Jun-xiang Yin, Yong-yan Wang, Beijing United Study Group on MCI of the Capital Foundation of Medical Developments. Guiding principles of clinical research on mild cognitive impairment (protocol). Journal of Chinese Integrative Medicine, 2008, 6(1): 9-14
[9] Bo Wang , Wei Yan , Li-hui Hou, Xiao-ke Wu. Disorder of Tiangui (kidney essence) and reproductive dysfunction in patients with polycystic ovary syndrome. Journal of Chinese Integrative Medicine, 2010, 8(11): 1018-1022
[10] 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