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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

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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]
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