| BackgroundAnterior Ischemic Optic Neuropathy (AION) is the most common optic nerve related visual loss disease in aged population(>50 years). Its pathological mechanism had been little explained for decades, and all the therapies are of weak capability to evidence based medicine, thus the only available treatment has been those non-specific ones such as steroid glucocorticoids. Though in China, Traditional Chinese Medicine has been supported by several clinical studies, all of them were neither supported by large sample, consciously proved evidence of EBM, nor delicately designed basic research. However, in recent years, we see a flurry increase researches both clinically or basically on AION. One of the main reasons is the establishment of an animal model which could highly mimic the nature process of the disease in human, which allows the pathophysiology of the disease could be gradually unveiled, and followed by a quantity of therapy research that right hitting the pathological mechanism. While to Traditional Chinese Medicine clinicians, the establishment of this animal model also opens up another vast field for the basic research of traditional therapies, and moreover, the controversy on the safety of some therapies could find a place to take the risk assessment study.As for the pathological mechanism, most researchers found that the main cause of retinal ganglion cells (RGCs) death is apoptosis induced by the stress of ischemia and anoxia of the optic nerve. And the loss of RGCs immediately results in the visual function loss.And at the same time, acupuncture has been widely accepted by most Traditional Chinese Medicine clinicians, and especially intraocular acupoints have become more and more popular and be considered to be of great value promoting in the clinic application. But the risks of intraocular acupoints are also known to most of clinicians and patients. How remarkable on earth is the effect of intraocular acupoints, and how big on earth is the value of promoting it universally? This is a question should be considered by most of the medical practitioners. Meanwhile, the effect of acupuncture interfering the process of apoptosis has also been widely studied. Thus this study, basing on the achievements of other studies, proposed that the mechanism of acupuncture treating AION may rely on the interference on mitochondrial path apoptosis that activated during the development of the disease.ObjectWe are going to examine the effect of acupuncture on mitochondrial path apoptosis of retinal ganglion cells on one hand, and on the other hand to determine the risk and benefit of intraocular acupoints through a rabbit model, which could provide a secondary opinion for clinicians.Methods12 New Zealand White (NZW) rabbits divided randomly in four groups (n=3):model group, intraocular acupoints group, extraocular acupoints group, intra-+extraocular acupoints, and induce AION on the right eyes of each animal utilizing photodynamic method, while take three of the left eyes randomly as control group. Apply acupuncture treatment to each group accordingly, that intraocular acupoints were Jingming (BL1), Chengqi (ST1), and Qiuhou (EX-HN7), extraocular acupoints were Cuanzhu (BL2), Yuyao (EX-HN4), Qiaoming, while intra-+extraocular acupoints combines the two groups. Cuanzhu (BL2), Yuyao (EX-HN4), Qiaoming were inserted about 1～1.5cm (0.3～0.5cun) by transvers insertion, while Jingming (BL1), Chengqi (ST1), Qiuhou(EX-HN7) were inserted about lcm (o.3cun) from the edge of the orbit, avoiding the eyeball. The needle was returned by 30min per day.3 days after treatment, all the animals were euthanized and the posterior segments of the eyes were harvested to prepare paraffin sections. Haematoxylin-eosin (HE) stain, immunohistochemical stain of Bax and Bcl-2 were applied. On this sections the thickness of inner retina were measured, and the numbers of positive RGCs of Bax and Bcl-2 were counted respectively, and finally the ratio of Bax/Bcl-2 were determined.3 sections of each animals were involved in statistical analysis (N=9).Results1) HE stain reveals the structural change of the retina, that oedema of the retina is obviously seen in model groups, and of which the structure is loss, while the intraocular acupoints group and extraocular acupoints group are not much better, however the intra-+ extraocular acupoints group see a tight structure. But concerning the thickness of inner retina, there’s no significance in comparing of each group.2) Bax positive RGCs count:a) Model group comparing with control group the p<0.05, is of significant difference, and the model group counts more Bax(+) RGCs than control group; b) Comparing with model group, all the treatment groups are of no significant differences; c) Comparing with the control group, extraocular acupoints group and intra-+ extraocular acupoints group are of significant differences, and the control group counts less Bax(+) RGCs then the two groups, while intraocular acupoints is of no significant difference; d) All the treatment groups comparing between each other see no significant differences, but there is a trend:model group> intra-+extraocular acupoints group> extraocular acupoints group> intraocular acupoints group> control group.3) Bcl-2 positive RGCs count:a) Model group comparing with control group the p<0.01, is of significant difference, and the model group counts less Bcl-2(+) RGCs than control group; b) Comparing with model group, extraocular acupoints and intra-+extraocular acupoints both have the p<0.01, are of significant differences, and the model group counts less Bcl-2(+) RGCs than the two groups, while intraocular group is of no significant difference; c) Comparing with the control group, intraocular acupoints group and extraocular acupoints group both have the p<0.05, are of significant differences, and the control group counts more Bcl-2(+) RGCs than the two groups, while intra-+extraocular acupoints is of no significant difference; d) Comparing with the intraocular acupoints group, the extraocular acupoints group and intra-+extraocular acupoints group both have the p<0.05, are of significant differences, and the intraocular acupoints group counts less Bcl-2(+) than the two groups; e) extraocular acupoints group comparing with intra-+extraocular acupoints group is of no significant difference; f) The trend is:control group> intra-+extraocular acupoints group> extraocular acupoints group> intraocular acupoints group> model group.4) Ratio of Bax/Bcl-2:a) Model group comparing with control group the p<0.01, is of significant difference, and the ratio of Bax/Bcl-2 of the model group is higher than the control group; b) Comparing with model group, extraocular acupoints and intra-+extraocular acupoints both have the p<0.01, are of significant differences, and the ratio of the model group is higher than the two groups, while intraocular group is of no significant difference; c) Comparing with the control group, intraocular acupoints group and extraocular acupoints group both have the p<0.05, are of significant differences, and the ratio of control group is lower than the two groups, while intra-+extraocular acupoints group is of no significant difference; d) Comparing with the intraocular acupoints group, the extraocular acupoints group and intra-+extraocular acupoints group both have the p<0.05, are of significant differences, and the ratio of intraocular acupoints group is higher than the two groups; e) extraocular acupoints group comparing with intra-+extraocular acupoints group is of no significant difference, f) The trend is:model group> intraocular acupoints group> extraocular acupoints group> intra-+extraocular acupoints group> model group.Conclusion1) Under the condition of this experiment, both intra-+extraocular acupoints group can decrease the Bax/Bcl-2 ratio, up to an extent they supressed the upper reaction of mitochondrial path of apoptosis;2) Although the exclusive intraocular acupoints makes no statistical significance to the model group, it can enhance the effect on decreasing the Bax/Bcl-2 when combining with the extraocular acupoints group;3) Generally, the three treatment groups show such a trend:intra-+extraocular acupoints group> extraocular acupoints group> intraocular acupoints group, suggesting that the combination of intra- and extraocular acupoints have the best effect on decreasing the Bax/Bcl-2. |
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