| ObjectiveObserve the effect of acupuncture with intraocular acupoints and extraocular acupoints on the apoptotic cells to verify the necessity of the selection of the acupoints in intraocular acupoints.Thus Enhance the scientific connotation of acupuncture for the treatment of non arteritis anterior ischemic optic neuropathy.Method1.Rabbit model of non-arteritic Anterior Ischemic Optic neuropathyA total of 3 New Zealand white rabbits were randomly divided into 3 groups,1 in each group.Chose the right eye as experimental eye.Using Rose Bengal as photosensitizer,NAION model of rabbit eyes was induced by green light with wavelength 532nm,power 75mW and spot diameter 1000um.40s,60s and 80s were irradiated respectively according to differrent groups.The model was evaluated by slit lamp and FFA one day after the model was finished.2.Preparation of NAION acupuncture modelA total of 20 New Zealand white rabbits were selected,and the right eye was used as the experimental eye.With Bengal rose as a photosensitizer,a model of non arteritis anterior ischemic optic neuropathy in rabbit eyes was induced by green light with wavelengths of 532nm,power 75mW,and the diameter of the spot 1000um.20 New Zealand rabbits were randomly divided into four groups,with 5 rats in each group:model group,intraocular needle group,extraocular needle group,intraocular and extraocular needle group,and then randomly selected 5 left eyes as control group.The acupuncture points in the intraocular acupuncture group were Jingming(BL1),Chengqi(ST1),and Qiuhou(EX-HN7),extraocular acupoints were Cuanzhu(BL2),Yuyao(EX-HN4),Qiaoming,while intraocular and extraocular acupoints combines the two groups.One day after the modeling,three acupuncture groups began acupuncture intervention,once a day,each time retaining 30min for 3 days,the model group and the normal group did nohing.After the treatment,all the animals were sacrificed to obtain4 the specimens of the posterior segment of the eye.The paraffin sections were made to detect the apoptosis of the cells by artificial counting cell apoptosis of TUNEL fluorescence labeling and the expression of Caspase-3 and active Caspase-3 by immunohistochemistry were detected by semi quantitative analysis of the photo density with image of Image Pro Plus 6.Result1.Preparation of NAION rabbit eye model:NAION animal model of rabbit eyes was established by laser inducing Rose Bengal,Under the same laser parameters,after different laser irradiation time of 40s,60s and 80s.As can be seen from the slit lamp examination,all three kinds of laser irradiation time can lead to upper optic disk edemaand the boundary is not clear.Fluorescein fundus angiography(FFA)showed low fluorescence in optic disc at early stage,the high fluorescence or background fluorescence was enhanced in the late stage,and the 40s lesion of fluorescence leakage was light,while the 80s was more serious,and the fluorescence leakage occurred in the early stage.Taking 60s as the final parameter of NAION model.2.Cell apoptosis count by TUNEL:(1)Compared with the normal group and the three acupuncture groups,the model group had significant difference in P=0.000<0.01,and the number of cell apoptosis in the model group was higher than that in the normal group and the three acupuncture groups;(2)There was no significant difference between the normal group and intraocular+extraocular acupuncture group(P=0.225>0.05).The mean number of apoptosis in the normal group was lower than that in the intraocular+ extraocular acupuncture group.There was significant differences between the extraocular acupuncture group and the normal group(P=0.015<0.05)and There was significant differences between the intraocular acupuncture group and the normal group(P=0.045<0.05).The number of apoptosis in normal group was lower than that in extraocular acupuncture group and intraocularl acupuncture group.(3)The mean number of apoptotic count showed that the intraocular+extraocular group<intraocular group<extraocular group,but there was no statistical difference between the three acupuncture groups with P>0.05 in each two groups.3.Positive expression of Caspase-3:(1)the model group compared with the normal group and the three acupuncture groups,P>0.05,there are no statistical difference;(2)the normal group compared with the three acupuncture groups,P>0.05,there is no statistical difference;(3)There was no statistical difference between the three acupuncture groups with P>0.05 in each two groups.4.Positive expression of active Caspase-3:(1)There were significantly statistical difference between model group and normal group,model group and intraocular+extraocular acupuncture group(P<0.01),there was statistical difference between model group and intraocula acupuncture group(P<0.05)and the expression of active Caspase-3 in model group was higher than that in the normal group,intraocular+extraocular acupuncture group and intraocula acupuncture group.There was no statistical difference between the model group and the extraocular acupuncture group(P>0.05),but the mean value of the model groups was higher than that of the extraocular acupuncture group;(2)The expression of active Caspase-3 in the normal group was lower than that in the three acupuncture groups(P<0.01),and the expression of active Caspase-3 in the normal group was lower than that in the three acupuncture groups.(3)There was no significant difference among the three acupuncture groups(P>0.05),and there was no significant difference between the intraocular acupuncture group,the extraocular group and the intraocular+extraocular group(P>0.05),Compared the mean values of three groups,extraocular group>intraocular group>intraocular+extraocular group;There was statistical difference between the extraocular group andintraocular+extraocular group(P=0.027<0.05).Conclusion1.NAION animal model was induced by photodynamic method.Under the same laser parameters,40s,60s and 80s laser irradiation time could be successfully used to prepare rabbit eye NAION animal model.The lesions in 40s were lighter,in 80s were more serious,and those in 60s were the best.60s irradiation time was used as the final parameter of NAION model,to provide model basis for NAION related experimental study of rabbit eyes.2.The intervention effect of acupuncture on apoptotic factor Caspase-3 needs to be further clarified,and acupuncture can exert its anti-apoptotic effect by down-regulating the active fragment active Caspase-3 downstream of Caspase-3,and then exerting its anti-apoptotic effect.According to the effect of reducing the expression of active Caspase-3 by different points selection,the effect of acupoint of intraocular+extraocular was the most best.3.Acupuncture intervention of NAION can play an anti-apoptosis role.And the intraocular and extraocular acupoint group was the best,which showed that the inhibition of optic nerve apoptosis was stronger after combination of intraocular+extraocular acupoint,suggesting that the method of selecting intraocular+extraocular acupoint is more beneficial to the recovery of the disease. |
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