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Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (9): 842-847.doi: 10.3736/jcim20090909

• Original Experimental Research • Previous Articles     Next Articles

Changes of mitochondrial apoptosis in spinal cord ischemia-reperfusion injury and the effects of Herba Erigerontis Breviscapi Injection preconditioning intervention in rabbits

 Bin Chenga,Xin Wangb,Lei Wanga,Kai Caoa,Lei Duanc,Feng-tao Lia   

  1. a Third Department of Orthopaedics, Second Hospital of Xi5an Jiao Tong University School of Medicine, Xi5an 710004, Shaanxi Province, China
    b Department of Orthopaedics, Affiliated Hospital, Employee Medical College, Xinzheng 451191, Henan Province, China
    c Department of Orthopaedics, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine, Shanghai 200437, China
  • Received:2009-04-20 Accepted:2009-07-06 Online:2009-09-20 Published:2009-09-15
  • Contact: Bin Cheng E-mail:drcheng2006@sina.com

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Objective

To investigate the mechanisms of mitochondrial apoptosis in spinal cord ischemia-reperfusion injury and the effects of Herba Erigerontis Breviscapi Injection preconditioning intervention.
Methods

Sixty Japanese rabbits were divided into sham-operated group, ischemia group, ischemia-reperfusion group (1, 6, 24 and 48 h), and Herba Erigerontis Breviscapi Injection group (1, 6, 24 and 48 h). Clamping the abdominal aorta was used to construct the rabbit model of spinal cord ischemia-reperfusion injury. The rabbits in the ischemia-reperfusion group and the Herba Erigerontis Breviscapi Injection group underwent reperfusion for 1, 6, 24, 48 h respectively after fifty-minute ischemia. The rabbits in the Herba Erigerontis Breviscapi Injection group were administered with Herba Erigerontis Breviscapi Injection at 9 mg/kg 30 minutes before ischemia. Rate of apoptotic cells was measured by flow cytometry; contents of caspase-9 and apoptosis-inducing factor (AIF) in cytoplasm and serum were measured by enzyme-linked immunosorbent assay.
Results

Compared with the sham-operated group and the ischemia group, the rates of apoptotic cells, the contents of caspase-9 and AIF in cytoplasm were increased at all time points after reperfusion, and the contents of caspase-9 and AIF in serum were decreased after 1 h and 6 h reperfusion, and increased after 24 h and 48 h reperfusion in the ischemia-reperfusion group. Herba Erigerontis Breviscapi Injection intervention could decrease the rate of apoptotic cells and the contents of caspase-9 and AIF in cytoplasm and serum as compared with those in the ischemia-reperfusion group, and the effects appeared after 1 h reperfusion.
Conclusion

The apoptosis of nerve cells after spinal cord ischemia-reperfusion is related to the mitochondrial pathways. Herba Erigerontis Breviscapi Injection can inhibit nerve cell apoptosis by decreasing the contents of caspase-9 and AIF in cytoplasm and serum.

Key words: Mitochondria, Herba Erigerontis Breviscapi Injection, Apoptosis, Ischemia-reperfusion injury, Rabbits

Figure 1

Rates of apoptosis of nerve cells in spinal cord measured by flow cytometry A: Sham-operated group; B: Ischemia group; C: Ischemia-reperfusion group, 1 h reperfusion; D: Ischemia-reperfusion group, 6 h reperfusion; E: Ischemia-reperfusion group, 24 h reperfusion; F: Ischemia-reperfusion group, 48 h reperfusion; G: Herba Erigerontis Breviscapi Injection group, 1 h reperfusion; H: Herba Erigerontis Breviscapi Injection group, 6 h reperfusion; I: Herba Erigerontis Breviscapi Injection group, 24 h reperfusion; J: Herba Erigerontis Breviscapi Injection group, 48 h reperfusion."

Table 1

Rates of apoptosis of nerve cells in spinal cord ($\bar{x}$±s, %)"

Group Time n Normal cells Apoptotic cells
Sham-operated 5 81.72±1.51 17.24±1.30
Ischemia 50 min 6 48.60±1.92* 42.82±1.44*
Ischemia-reperfusion 1 h 6 38.53±2.18*△ 57.34±1.95*△
6 h 6 23.41±1.64*△ 70.52±1.93*△
24 h 6 14.97±0.53*△ 81.03±0.47*△
48 h 6 32.87±1.18*△ 64.04±1.69*△
Herba Erigerontis Breviscapi Injection 1 h 6 45.38±0.96▲ 45.85±1.14▲
6 h 6 30.50±0.74▲ 65.32±1.44▲
24 h 6 27.89±1.85▲ 64.33±0.50▲
48 h 6 46.67±1.30▲ 41.47±1.19▲

Table 2

Contents of AIF and caspase-9 in cytoplasm of spinal cord and serum ($\bar{x}$±s)"

Group Time n Cytoplasm Serum
AIF (ng/L) Caspase-9 (μg/L) AIF (ng/L) Caspase-9 (μg/L)
Sham-operated 5 211.21±10.84 1.92±0.16 788.52±23.87 3.95±0.20
Ischemia 50 min 6 242.11±16.49 2.38±0.36 1 057.85±58.85* 8.40±0.15*
Ischemia-reperfusion 1 h 6 350.93±16.86*△ 4.30±0.29*△ 863.29±15.83△ 5.69±0.34*△
6 h 6 731.72±15.50*△ 5.57±0.28*△ 931.16±19.21*△ 6.38±0.39*△
24 h 6 540.38±8.64*△ 5.31±0.37*△ 1 606.99±40.92*△ 9.31±0.18*△
48 h 6 344.76±20.65*△ 4.05±0.25*△ 1 157.64±28.88*△ 8.41±0.15*
Herba Erigerontis Breviscapi Injection 1 h 6 339.50±20.54 3.35±0.14▲ 802.24±17.62 4.77±0.11▲
6 h 6 449.01±23.75▲ 3.81±0.21▲ 800.08±18.63▲ 5.01±0.27▲
24 h 6 392.94±14.47▲ 3.51± 0.28▲ 920.19±29.94▲ 6.33±0.39▲
48 h 6 296.91±8.18▲ 3.06±0.24▲ 809.06±24.02▲ 5.36±0.17▲
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