| 摘要: | 針灸已廣泛的應用於臨床治療疾病如癲癇、中風等，針灸對於神經細胞損傷是否具有神經細胞保護作用或促進神經細胞再生至今仍然不明。因此，本研究的目的是探討針刺對神經細胞損傷的效用。我們將KA (kainic acid) 1μg/μl注入Sprague-Dawley（SD）大鼠右側海馬CA1上緣後，觀察3小時的大鼠癲癇發作行為，選擇癲癇發作行為Racine分類第四級以上，且wet dog shakes發作次數介於150到200次之大鼠，然後隨機分為PBS (Phosphate Buffered Saline)組、KA組、假電針組和電針組（2Hz電針；右率谷和天衝穴），每組6隻。電針刺激是從KA注射後的第二天開始到第六天，每天一次10分鐘。KA注射後第7天，將大鼠犧牲取腦做成切片，分別以HE (Hemeatoxylin & Eosin) 染色、 NeuN (Neuron-specific nuclear protein) 和BDNF (Brain-derived neurotrophic factor) 免疫組織化學染色觀察大鼠海馬區域的組織結構、和神經細胞。結果顯示KA注射後海馬區域的組織構造破壞，而假電針和電針能減少這些破壞。海馬CA1區域的NeuN染色陽性細胞數，PBS組是188±9.32大於KA組的17±3.71，假電針組的59±3.20和電針組的128±5.42 (all p < 0.05)；電針組的NeuN陽性染色細胞數大於KA組和假電針組（both p < 0.05）；假電針的NeuN染色陽性細胞數大於KA組（p < 0.05）。海馬CA3區域的NeuN染色陽性細胞數，PBS組是137±5.50大於KA組的6±2.44，假電針組的30±2.75和電針組的50±2.86 (all p < 0.05)；電針組的NeuN陽性染色細胞數大於KA組和假電針組（both p < 0.05）；假電針的NeuN染色陽性細胞數大於KA組（p < 0.05）；海馬CA1區域的BDNF染色陽性細胞數，電針組是301±5.50大於PBS組的26±3.87，KA組的6±1.77和假電針組是150±5.85 (all p < 0.05)；假電針組的BDNF染色陽性細胞數於KA組和PBS組（both p < 0.05）；PBS組的BDNF染色陽性細胞數大於KA組（p < 0.05）；CA3區域的BDNF染色陽性細胞數，電針組則是53±3.11大於PBS組的8±1.97，KA組的3±1.34和假電針組的32±3.08 （all p <0.05）；假電針針組的BDNF染色陽性細胞數於KA組和PBS組（both p < 0.05）；PBS組的BDNF陽性細胞數大於KA組（p < 0.05）。
結論是2Hz電針施予率谷和天衝穴對KA治療誘發神經細胞損傷有神經細胞的保護作用。
Acupuncture had been widely used to treat disease, such as epilepsy, stroke etc. However, effect of acupuncture on neuroprotection and enhancing neuronal regeneration remain unclear. Therfore, the purpose of the present study was to investigate the effect of acupuncture on neuronal cell damage, we injected KA (kainic acid ) 1μg/μl to rat right hippocampal CA1’s edge, and then observed these rats’ behaviors while epilepsy attacked. We selected these rats which fitted above the 4th stage of Racine’s classifications and 150-200 times of wet-dog shakes after KA injection. Afterwards, screened rats were randomly assigned to PBS group, KA group, sham group and EA (electro-acupuncture) group (2Hz; GB8 and GB9 acupoints on the right side), there were six rats in each group. We sacrificed rats and removed brain tissue out of their heads and made brain sections 7th day after KA administration. We used HE stain, and NeuN and BDNF immunohistochemistry stain respectively to observe rats’ hippocampal tissue and neurons. The results showed that KA could destroy hippocampal tissue, but sham and elector-acupuncture reduced these damages. In the CA1 region of the hippocampus, the count of NeuN stain positive cells was 188±9.32 in PBS group greater than 17±3.71 in KA group, 59±3.20 in sham group, and 128±5.42 in EA group (all p < 0.05); the count of NeuN stain positive cells in EA group was greater than that in KA group and sham group (both p < 0.05); the count of NeuN stain positive cells in sham group was greater than that in KA group ( p < 0.05). In the CA3 region of the hippocampus, the count of NeuN stain positive cells was 137±5.50 in PBS group greater than 6±2.44 in KA group, 30±2.75 in sham group, and 50±2.86 in EA group (all p < 0.05); the count of NeuN stain positive cells in EA group was greater than that in KA group and sham group (both p < 0.05); the count of NeuN stain positive cells in sham group was greater than that in KA group ( p < 0.05). In the CA1 region of the hippocampus, the count of BDNF stain positive cells was 301±5.50 in EA group greater than 26±3.87 in PBS group, 6±1.77 in KA group, and 150±5.85 in sham group (all p < 0.05); the count of BDNF stain positive cells in sham group was greater than that in KA group and PBS group (both p < 0.05); the count of BDNF stain positive cells in PBS group was greater than that in KA group ( p < 0.05). In the CA3 region of the hippocampus, the count of BDNF stain positive cells was 53±3.11 in EA group greater than 8±1.97 in PBS group, 3±1.34 in KA group, and 32±3.08 in sham group (all p < 0.05); the count of BDNF stain positive cells in sham group was greater than that in KA group and PBS group (both p < 0.05); the count of BDNF stain positive cells in PBS group was greater than that in KA group ( p < 0.05).
Based on the results of mention-above, we infer that 2Hz EA on GB8 and GB9 acupoints have neuroprotection on kainic acid-induced hippocampal neuronal damage. |
