基于炎症反应探讨补阳还五汤对AD模型小鼠的神经保护作用及机制

To Explore the Neuroprotective Effect and Mechanism of Buyang Huanwu Decoction on AD Model Mice Based on Inflammatory Response

目的探讨补阳还五汤对阿尔茨海默病(AD)模型小鼠的脑神经保护作用及其机制。方法将60只AD模型小鼠分为补阳还五汤高、中、低剂量组(37.07 g/kg、18.54 g/kg、9.27 g/kg)、脑复康组(吡拉西坦0.62 g/kg)和模型组(等体积生理盐水),另选12只C57小鼠为假手术组(等体积生理盐水)。以上各组连续给药20 d,末次给药后断头取脑,然后采用HE法观察各组脑组织形态学异常;ELISA法检测炎性相关因子TNF-α和IL-6的表达;IHC法检测凋亡相关因子Bcl-2和Bax的表达;采用WB法检测信号通路p38MAPK/NF-κB的蛋白表达。结果补阳还五汤组的神经细胞排列较为规整,细胞炎性肿胀情况减轻,尼氏小体有所增加,空泡化减少;与模型组相比,补阳还五汤中、高剂量组TNF-α和IL-6的表达明显下降(P<0.01),抗凋亡因子Bcl-2表达上升,促调亡因子Bax表达下降(P<0.01、P<0.05),NF-κB和p38MAPK蛋白的表达也都出现了下降现象(P<0.01)。结论补阳还五汤可以抑制AD模型小鼠脑组织炎症反应,减少神经细胞凋亡,并推测此神经保护作用可能与调控炎症通路p38MAPK/NF-κB有关。

Objective To explore the neuroprotective effect of Buyang Huanwu decoction on Alzheimer's disease(AD)model mice and its mechanism.Methods Sixty AD model mice are divided into the high,middle and low dose groups of Buyang Huanwu decoction(37.07 g/kg,18.54 g/kg,9.27 g/kg),the naofukang group(piracetam 0.62 g/kg)and the model group(equal volume normal saline).Twelve C57 mice are selected as the sham operation group(equal volume normal saline).The above groups are continuously administered for 20 days.After the last administration,the head is decapitated and the brain is removed.Then the abnormality of brain tissue morphology is observed by HE method.The expression of inflammatory related factor TNF-αand IL-6 is detected by ELISA method.The expression of apoptosis related factor Bcl-2 and Bax is detected by IHC.The protein expression of signal pathway p38MAPK/NF-κB is detected by WB method.Results In the Buyang Huanwu decoction group,the arrangement of nerve cells is more regular,the inflammatory swelling of cells is reduced,the Nissl body is increased,and the vacuolization is decreased.Compared with the model group,in Buyang Huanwu decoction middle and high dose group,the expression of TNF-αand IL-6 are significantly lower(P<0.01),the expression of anti-apoptotic factor Bcl-2 increased and the expression of pro-apoptotic factor Bax decreased(P<0.01,P<0.05),the expression of NF-κB and p38MAPK decreased(P<0.01).Conclusion Buyang Huanwu decoction can inhibit the inflammatory response in the brain tissue of AD model mice,thus reducing the apoptosis of nerve cells.It is speculated that this neuroprotective effect might be related to p38MAPK/NF-κB,which regulates the inflammatory pathway.