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Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (1): 25-33.doi: 10.3736/jcim20090104

• Original Clinical Research • Previous Articles     Next Articles

  • Herbs for calming liver and suppressing liver-yang in treatment of migraine with hyperactive liver-yang sydrome and its effects on lymphocyte protein expression: a randomized controlled trial

 Guang-wei Zhonga, Wei Lia, Yan-hong Luob, Guo-lin Chena, Zhen-jia Yia ,Jian-jun Hua, Ying Zhanga, Yao-hui Yina, Le Zhangc   

  1. a Research Institute of Integrated Traditional Chinese and Western Medicine, Xiangya Hospital, Central South University, Changsha, Hunan Province 410008, China
    b Department of Emergency, Xiangya Hospital, Central South University, Changsha, Hunan Province 410008, China
    c Department of Neurology, Xiangya Hospital, Central South University, Changsha, Hunan Province 410008, China
  • Received:2008-07-30 Accepted:2008-08-12 Online:2009-01-20 Published:2009-01-15

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Objective

To observe the efficacy of herbs for calming liver and suppressing liver-yang in treatment of migraine patients with hyperactivity of liver-yang syndrome and to investigate its effects on the lymphocyte protein expression. This approach may lay a foundation for the further investigation of pathogenic mechanisms in migraine with hyperactive liver-yang sydrome and the curative mechanisms of calming liver and suppressing liver-yang treatment.
Methods

A total of 32 migraine patients with hyperactivity of liver-yang syndrome were randomly divided into treatment group (16 cases) and control group (16 cases). The patients in the treatment group were treated with herbs for calming liver and suppressing liver-yang in accordance with traditional Chinese medicine (TCM) theory and the patients in the control group were treated with Flunarizine Capsules for two courses of treatment. The therapeutic effects, the score of TCM symptom and the changes of headache attack were observed in both groups before and after the treatment. The side effects were also observed in both groups. The level of differential protein expression was analyzed by two-dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS).
Results

The herbs for calming liver and suppressing liver-yang had better effects on headache improvement than the Flunarizine Capsules (P<0.01). The cure rates in the treatment and control groups were 87.5% and 75.0% respectively. Vertigo, restlessness and tantrum, and prosopo-heat in the treatment group were also improved as compared with those in the control group (P<0.05). After treatment, the score of TCM symptom in the two groups were all decreased (P<0.01), and there was a significant difference between the treatment group and the control group (P<0.01). The herbs for calming liver and suppressing liver-yang had no side effects. The average protein spots in the blood lymphocyte of normal people, migraine patients with hyperactivity of liver-yang syndrome in the treatment group before and after the treatment were (534±42), (552±54) and (529±55) spots respectively. Six down-regulated protein expressions and 14 up-regulated protein expressions were obtained in the treatment group. Four strengthened protein expressions in the six down-regulated proteins and 11 low protein expressions in the 14 up-regulated proteins were also obtained after treatment. Ten of the total 12 differential protein spots were successfully identified by MALDI-TOF-MS. The functions of these proteins were involved in metabolism, energy generation, transportation, antioxidation, signal transduction and immune, etc. According to information provided by NCBI and MSDB database, there were some proteins closely related to migraine with hyperactivity of liver-yang syndrome, such as peroxiredoxin 2, heat shock protein 27 and annexin A1.
Conclusion

 Herbs for calming liver and suppressing liver-yang is effective in treating migraine, and can improve TCM symptoms. The effects on migraine patients with hyperactivity of liver-yang syndrome may be related to regulating the blood lymphocyte protein expression.

Key words: Calming the liver, Suppressing liver-yang, Liver-yang hyperactivity, Proteomics, Migraine disorder, Randomized controlled trials

Table 1

Attack of headache in the treatment and control groups (x±s)"

Group n Frequency (attacks/month) Persistence time (h)
Control
Before treatment 16 3.5±0.58 5.48±1.43
After treatment 16 1.8±0.34** 2.43±0.46**
Treatment
Before treatment 16 3.4±0.56 5.62±1.36
After treatment 16 1.2±0.37**△ 1.87±0.69**△

Table 2

Therapeutic effects on headache in the treatment and control groups [Cases (%)]"

Group n Cure Obvious effect Improvement No improvement Total obvious effect
Control 16 3 (18.75) 6 (37.50) 3 (18.75) 4 (25.00) 12 (75.00)
Treatment 16 5 (31.25) 7 (43.75) 2 (12.50) 2 (12.50) 14 (87.50)△△

Table 3

Main symptoms of hyperactivity of liver-yang syndrome in the treatment and control groups [Cases]"

Group n Headache Dry mouth Vertigo Restlessness and tantrum Prosopo-heat
Control
Before treatment 16 16 14 10 13 12
After treatment 16 4** 6** 6** 9** 8**
Treatment
Before treatment 16 16 13 11 14 13
After treatment 16 3** 7** 3**△ 6**△ 6**△

Table 4

Score of traditional Chinese medicine symptom in the treatment and control groups (x±s)"

Group n Score of traditional
Chinese medicine symptom
Control
Before treatment 16 20.82±7.28
After treatment 16 10.73±3.92**
Treatment
Before treatment 16 21.64±6.65
After treatment 16 7.21±3.18**△△

Figure 1

Two-dimensional electrophoresis map of human in the normal and treatment groups A: Normal group; B: Before treatment in the treatment group; C: After treatment in the treatment group."

Table 5

Relative intensity of protein expression in the normal and treatment groups"

SSP Normal group
(x±s, n=10)		 Before treatment in the
treatment group (x±s, n=10)		 Ratio After treatment in the treatment
group(x±s, n=10)		 Ratio
1016 784.1±24.5 2 124.6±26.5 2.71 937.2±38.2 0.44
1207 368.3±28.6 981.2±17.7 2.66 458.1±22.4 0.47
1318 16.2±3.5 171.6±21.2 10.60 62.5±19.3 0.36
1420 35.6±4.2 1 226.8±54.9 34.46 367.3±43.4 0.29
1625 109.8±35.7 763.5±58.5 6.95 654.7±61.4 0.85
1801 673.7±56.6 1 532.7±116.1 2.28 1 738.5±88.9 1.13
2019 484.7±43.6 1 491.1±83.6 3.08 586.1±43.6 0.39
2013 123.1±15.1 4 282.5±238.1 34.76 576.2±47.4 0.13
2209 357.2±23.2 1 091.5±71.1 3.06 735.6±57.7 0.67
2259 263.4±22.5 2 363.4±128.1 8.97 472.3±36.9 0.20
4204 1 281.6±112.2 2 831.4±121.6 2.21 1 153.8±138.2 0.41
5606 140.3±12.8 362.4±42.9 2.58 116.7±19.3 0.32
5625 20.6±4.1 1 517.7±111.5 73.67 367.9±41.6 0.24
7234 24.5±5.4 231.3±31.8 9.44 111.8±14.3 0.48
2225 1 073.5±42.3 461.9±34.8 0.43 964.2±67.5 2.09
2115 692.3±56.5 301.6±47.6 0.44 838.5±56.9 2.78
2350 992.9±65.6 178.3±26.5 0.18 392.7±43.6 2.22
2313 934.8±73.8 304.3±32.7 0.33 484.3±21.3 1.59
2374 2 200.7±87.1 387.1±65.5 0.18 326.8±30.1 0.84
2439 797.8±41.5 169.7±31.1 0.21 527.6±65.4 3.11

Figure 2

Peptide mass fingerprinting of sample 7 by MALDI-TOF-MS analysis"

Figure 3

Mascot search results of peptide mass fingerprinting of sample 7 in Swissport"

Table 6

Proteins identified by Mascot software"

Sample Database ID Protein name Molecular
weight		 Isoelectric
point		 Matched
peptides		 Sequence
coverage		 Expression
1 Q71V49_HUMAN Alcohol dehydrogenase 3 1 217 9.52 5 30% ↓
2 S26788 Immunoglobulin heavy
chain Ⅴ region		 12 472 8.01 7 33% ↑
3 E980237 Heat shock protein-27 22 427 7.83 5 78% ↑
4 DEF4_HUMAN Neutrophil defensin
4 precursor		 10 839 8.31 5 40% ↑
5 MAGE2_HUMAN Melanoma-associated
antigen E2		 12 344 6.63 4 35% ↑
6 Q8IUS2_HUMAN LOC339483 protein 11 303 6.67 7 33% ↑
7 P04083_HUMAN Annexin-A1 38 583 6.64 5 59% ↑
8 AAB06412_HUMAN Immunoglobulin heavy
chain variable region (Fragment)		 13 424 8.57 3 41% ↑
9 P35704_HUMAN Peroxiredoxin-2 21 956 5.34 8 49% ↑
10 Q9UC96_HUMAN MU class glutathione
S-transferase (Fragment)		 3 544 5.50 5 40% ↓
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