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Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (10): 940-945.doi: 10.3736/jcim20091006

• Original Experimental Research • Previous Articles     Next Articles

Protective effects of Ginkgo biloba extract on morphology and function of retinal ganglion cells after optic nerve transection in guinea pigs

Zheng-gao Xiea,Xing-wei Wub,Chao-rong Zhuanga,Fang Chena,Zheng Wanga,Ya-kun Wanga,Xin Huaa   

  • Received:2009-06-09 Accepted:2009-08-05 Online:2009-10-20 Published:2009-10-15
  • Contact: Zheng-gao Xie E-mail:zgxie87@163.com

Objective

To investigate the effects of Ginkgo biloba extract (EGb 761) on the morphology and function of retinal ganglion cells (RGC) in guinea pigs with optic nerve transection.
Methods

Seventy-five albino guinea pigs were randomly divided into five groups: normal control group, sham-operated group, untreated group, normal saline group and EGb 761 group. No operation was performed in the normal control group. Optic nerve was merely exposed in the sham-operated group, but transected at 1.0 mm from posterior pole of the eye ball in the untreated, normal saline and EGb 761 groups. Guinea pigs in the EGb 761 group or the normal saline group received daily intraperitoneal injection of EGb 761 (100 mg/kg) or corresponding volume of normal saline from 7 days before experiment to 28 days after experiment. Three guinea pigs in each group were sacrificed for apoptosis assay (TUNEL method) of RGC. Pattern electoretinograms (PERGs) were recorded 14 and 28 days after transection, respectively. At the end of the examination, six guinea pigs were killed for histological examination and RGC count.
Results

No TUNEL-positive cells were observed in the normal control, sham-operated and EGb 761 groups, but there were TUNEL-positive cells in the untreated group and the normal saline group. The numbers of RGCs in the untreated and normal saline groups were less than those in the normal control and sham-operated groups at 14 days or 28 days (P<0.05). Although the number of RGCs in the EGb 761 group was less than those in the normal control and sham-operated groups (P<0.05), it was more than those in the untreated and normal saline groups (P<0.05). N95 amplitude in EGb 761 group was higher than those in the untreated and normal saline groups (P<0.05) and close to those in the normal control and sham-operated groups (P>0.05) at 14 days or 28 days. The number of RGCs was positive correlated to N95 amplitude (r=0.859, P=0.001 5).
Conclusion

EGb 761 can inhibit the apoptosis of RGCs in guinea pigs after optic nerve transection, thus protect the morphology and function of RGCs.

Key words: Ginkgo biloba extract, Optic nerve lesion, Retinal ganglion cell, Pattern electroretmogram, Neuroprotective effect, Guinea pigs

Figure 1

Effects of EGb 761 on RGC apoptosis checked by TUNEL (Fluorescein microscopy, ×400) GCL: Ganglion cell layer; INL: Inner nuclear layer; ONL: Outer nuclear layer; TUNEL: DeadEnd fluorometric terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling; PI: Propidium iodide. A: Normal control group, no TUNEL positive RGCs; B: Sham-operated group, no TUNEL-positive RGCs in retina; C: Untreated group, a few TUNEL positive RGCs (white arrows) presented in retina; D: Normal saline group, a few TUNEL-positive RGCs (white arrows) presented in retina; E: EGb 761 group, no TUNEL-positive RGCs presented in retina."

Figure 2

Effects of EGb 761 on retinal structure observed by HE staining (Light microscopy, ×400) GCL: Ganglion cell layer; INL: Inner nuclear layer; ONL: Outer nuclear layer. A: Normal control group. There was no vascular structure in retina, and cubic RGCs arrayed regularly and the nerve fibers layer (NFL) of retina was thick. B: Sham-operated group. The number of RGCs was close to that in normal control group. C: Untreated group. The number of RGCs and the thickness of NFL were obviously less than those in normal control group. D: Normal saline group. RGCs were rare and NFL was very thin. E: EGb 761 group. The number of RGCs and the NFL were slightly less than those in normal control group. All cells white arrows pointed at were RGCs."

Figure 3

Survival index of RGC in each group 14 and 28 days after optic nerve transection A: Normal control group; B: Sham-operated group; C: Untreated group; D: Normal saline group; E: EGb 761 group. Data were represented as $\bar{x}$±s, n=6. *P<0.05, vs normal control group; △P<0.05, vs sham-operated group; ▲P<0.05, vs untreated group; □P<0.05, vs normal saline group."

Figure 4

Changes of N95 amplitude in each group 14 and 28 days after optic nerve transection A: Normal control group; B: Sham-operated group; C: Untreated group; D: Normal saline group; E: EGb 761 group. Data were represented as $\bar{x}$±s, n=6. *P<0.05, vs normal control group; △P<0.05, vs sham-operated group; ▲P<0.05, vs untreated group; □P<0.05, vs normal saline group."

Figure 5

PERG wave form in each group 28 days after optic nerve transection N35: Negative wave 35; P50: Positive wave 50; N95: Negative wave 95. A: Normal control group; B: Sham-operated group; C: Untreated group; D: Normal saline group; E: EGb 761 group."

Figure 6

Correlation of N95 amplitude with survival index of RGC"

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