Anti-apoptotic and neuroprotective effects of Acupuncture at LR3 and GB37 on focal vrain ischemic injury induced by Intraluminal Filament insertion in Rats : 太衝, 光明, 太衝配光明 刺鍼이 局所 腦虛血 모델 白鼠에서 抗細胞枯死 및 腦神經保護에 미치는 影響
초록/요약
足厥陰肝經의 太衝과 足少陽膽經의 光明은 임상에서 原絡配穴原理를 적용하여 肝風으로 인한 중풍초기 및 후유증 치료에 응용하고 있다. 하지만 이에 대한 효과 관찰 및 그 기전연구가 미진하여 본 연구에서는 Intraluminal Filament 揷入術에 의하여 유발된 白鼠의 虛血性 局所 腦損傷모델에 대하여 太衝 및 光明의 原絡配穴 鍼刺治療法이 腦神經의 抗細胞枯死와 保護效果에 미치는 影響을 관찰하였다. 局所 腦損傷은 中大腦動脈을 結紮하여 誘發하였으며, 각 군들은 정상군(Normal)과 局所 腦損傷 유발 후 무처치한 대조군(Control), 局所 腦損傷 유발 후 태충에 침자한 군(LR3), 局所 腦損傷을 유발 후 광명에 자침한 군(GB37), 局所 腦損傷 유발 후 태충과 광명에 자침한 군(LR3+GB37) 등 5군으로 구성되었으며, 腦神經의 抗枯死 및 保護效果는 TTC 염색, 해마에서의 mGluR5, Bax, Bcl-2, cytochrome c의 발현정도와 해마의 CA1부위를 면역화학염색하여 신경세포의 손실정도 및, ChAT, NGF, c-Fos의 발현들을 광학현미경으로 관찰하였다. 본 실험의 결과, 太衝 및 太衝과 光明에 刺鍼한 군에서 mGluR5, BCL-2 및 ChAT의 발현과 TTC와 Cresyl violet에 의하여 염색된 腦神經細胞의 濃度 유의하게 증가하였고, Bax 와 cytochrome c의 濃度와 Bax에 대한 Bcl-2 比率이 감소하였다. 위의 결과를 종합해보면 太衝 및 太衝과 光明에 刺鍼療法이 腦神經의 抗細胞枯死 및 保護效果에 영향을 주는 것으로 보아, 中風初期治療에 效果가 있는 것으로 觀察되었다.
more초록/요약
Objectives : Acupuncture using a source-connecting method is used as a controlling the medication for an early stroke in the Korean medicine. LR3 is the source acupoint of a liver meridian and GB37 is the connecting acupoint of a Gallbladder meridian. So this study is aims to investigate the anti-apoptotic and neuro-prtective effects of acupuncture on the focal ischemia induced by intraluminal filament insertion in rats. Materials and methods : The focal ischemia was induced by Intraluminal Filament insertion into middle cerebral artery. The animals were divided into five groups (n=15 in each group) : not ischemia induced and not treated group (Normal), the ischemia induced and not treated group (Control), the ischemia induced and acupunctured at LR3 group (LR3), the ischemia induced and acupunctured at GB37 group (GB37) and the ischemia induced and acupunctured at LR3 and GB37 group (LR3+GB37). The anti-apoptotic and neuroprotective effects of acupuncture are observed by mGluR5, Bax, Bcl-2, cytochrome c, TTC, Cresyl violet, ChAT-stain, NGF and c-Fos. Results : The infarction had progressively decreased in the LR3 group, GB37 group and LR3+GB37 group. The mGluR5, BCL-2, density of neurons stained by Cresyl violet and ChAT had increased in LR3 group and LR3+GB37group, The intensity of Bax, the ratio of Bax/Bcl-2, cytochrome c had decrease in LR3 group and LR3+GB37group. The density of NGF had only increased in LR3+GB37group. Conclusions : Our study suggests that acupuncture treatment at LR3, LR3 and GB37 shows anti-apoptotic and neuroprotective effects on cholinergic neuron in focal cerebral ischemia of the stroke in rats.
more목차
Contents = ⅰ
Abstract = ⅲ
Ⅰ. Introduction = 1
Ⅱ. Materials ＆ Methods = 3
1. Animals = 3
2. Groups and treatment = 3
3. Occlusion by Intraluminal Filament insertion = 3
4. Triphenyltetrazolium Chloride (TTC)-stain = 4
5. Isolation of total RNA and Reverse Transcription Polymerase Chain Reaction (RT-PCR) = 4
6. Western blotting = 5
7. Immunohistochemistry = 6
1) Preparation of tissues = 6
2) Cresyl violet stain = 7
3) Cholineacetyltransferase(ChAT), Nerve Growth Factor(NGF) and c-Fos = 7
8. Data Analysis = 8
Ⅲ. Results = 9
1. Infarction volume = 9
2. The expression of mGluR5 in the right hippocampus = 10
3. The expression of Bax and Bcl-2 in the right hippocampus = 11
1) The expression of Bax in the right hippocampus = 11
2) The expression of Bcl-2 in the right hippocampus = 12
3) The ratio of Bax/Bcl-2 in the right hippocampus = 13
4. The release of cytochrome c in the right hippocampus = 14
5. The density of Cresyl violet-stained neural cell in the right hippocampal CA1 = 15
6. The density of ChAT in the right hippocampal CA1 = 17
7. The density of NGF in the right hippocampal CA1 = 19
8. The expression of c-Fos in the right hippocampal CA1 = 21
Ⅳ. Discussion = 23
Ⅴ. Conclusion = 29
References = 31
국문초록(Korean abstract) = 35