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Journal of Chinese Integrative Medicine ›› 2011, Vol. 9 ›› Issue (9): 991-996.doi: 10.3736/jcim20110910

• Original Experimental Research • Previous Articles     Next Articles

Protective effects of Chinese herbal medicine Zuogui Pill on retina ganglion cells after optical nerve clipping injury in rats

Yong-qian Wang1(), Xiao-feng Li2, Xin Zhou3, Xin-quan Liu4, Wei-ping Wang1   

  1. 1. Department of Neurosurgery, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
    2. Institute of Spine Diseases, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
    3. Department of Pharmacy, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
    4. Department of Ophthalmology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
  • Received:2011-04-04 Accepted:2011-05-09 Online:2011-09-20 Published:2011-09-15
  • Contact: Wang Yong-qian E-mail:sunmoonciel@163.com

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Objective: To investigate the protective effects of Zuogui Pill (ZGP), a compound Chinese herbal medicine, on retina ganglion cells (RGCs) in rats after optical nerve clipping injury.

Methods: Optic nerves in Sprague-Dawley rats were crushed intraorbitally at 2 mm behind eyeball by using a forceps with a 140-gram chucking power for 30 s to induce unilateral injured model. The injured rats were fed with ZGP (ZGP group, 4.0 g/kg body weight each day) and equal volume of normal saline (injury group) respectively. Morphological changes in the retina were observed and the RGCs were quantified in the specimens. Nestin and glial fibrillary acidic protein (GFAP) expressions in the retina were detected by immunohistochemical fluorescence staining at different time points after nerve injury.

Results: After optical never crushing, the thickness of retina and the number of RGCs were significantly decreased in injury group as compared with the normal group. Nearly 50% of the total RGCs were disappeared at the 4th week in the injury group. The expression levels of nestin and GFAP in the retina were markedly increased after optical never crushing at the 2nd week and this lasted to the 8th week, and gradually decreased to the normal level at the 16th week. In contrast, the arrangement of retinal cells was regular in the ZGP group, and the number of survival RGCs was greater than that of the injury group. Meanwhile, the nestin and GFAP expressions in retina were significantly enhanced as compared with the injury group at all time points.

Conclusion: ZGP may have proctective effects on rat optic nerve and RGCs after contusion by promoting the nestin and GFAP expressions in Muller cells of the retina.

Key words: Zuogui Pill, drugs,Chinese herbal, retina, retinal ganglion cells, optic nerve injury, nestin, glial fibrillary acidic protein, rats

Table 1

Numbers of RGCs in retina at different time points after optic nerve crush in adult rats ($\overline{x}$±s)"

Group n Number of RGCs
1 week 2 weeks 4 weeks 8 weeks 16 weeks
Normal 5 40.13±5.42
Sham injury 5 40.07±4.65
Injury 3 31.27±3.57** 24.00±3.38** 19.93±2.15** 17.13±2.13** 15.00±2.51**
ZGP 3 34.60±3.79**△ 28.07±3.41**△△ 24.20±2.36**△△ 20.93±2.37**△△ 17.26±2.63**△

Table 2

IOD of nestin immunoreactivity in retina after optic nerve crush in adult rats ($\overline{x}$±s)"

Group n IOD of nestin
1 week 2 weeks 4 weeks 8 weeks 16 weeks
Normal 5 0.013±0.004
Sham injury 5 0.015±0.004
Injury 3 0.023±0.007** 0.028±0.005** 0.037±0.006** 0.029±0.006** 0.015±0.005
ZGP 3 0.029±0.007**△ 0.036±0.006**△△ 0.043±0.006**△△ 0.033±0.006**△△ 0.022±0.005**△△

Table 3

IOD of GFAP immunoreactivity in retina after optic nerve crush in adult rats ($\overline{x}$±s)"

Group n IOD of GFAP
1 week 2 weeks 4 weeks 8 weeks 16 weeks
Normal 5 0.012±0.005
Sham injury 5 0.013±0.007
Injury 3 0.024±0.008** 0.031±0.005** 0.037±0.006** 0.030±0.006** 0.019±0.005*
ZGP 3 0.031±0.008**△ 0.037±0.006**△△ 0.044±0.007**△△ 0.034±0.006**△ 0.024±0.006**△

Figure 1

Expressions of DAPI, nestin and GFAP in retina tissues of rats observed by immunofluorescence method A: Nestin expression in the normal group; B: GFAP expression in the normal group; C: Nestin expression in the injury group at the 4th week after injury; D: GFAP expression in the injury group at the 4th week after injury; E: Nestin expression in the Zuogui Pill group at the 4th week after injury; F: GFAP expression in the Zuogui Pill group at the 4th week after injury. Nestin expression was stained with green, NAPI with blue and GFAP with red. ZGP: Zuogui Pill; GFAP: glial fibrillary acidic protein; DAPI: 4′,6-diamidino-2-phenylindole; GCL: retinal ganglion cell layer; INL: inner nuclear layer; IPL: inner plexiform layer; ONL: outer nuclear layer. Arrows point to the positive processes in the IPL. Scale bar=50 μm."

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