经方小柴胡汤化裁方对阿尔茨海默病小鼠海马组织细胞炎性因子IL-1β、IL-6及p-p38 MARK、NF-κB表达水平的影响

Effects of Xiaochaihu Decoction on Expressions of IL-1β, IL-6, p-p38 MARK and NF-κB in Hippocampus of Alzheimer’s Disease Mice

目的观察经方小柴胡汤化裁方对阿尔茨海默病(AD)小鼠海马组织细胞炎性因子IL-1β、IL-6及p-p38 MARK、NF-κB表达水平的影响,探讨其机制。方法选取SPF级小鼠30只,随机分成6组,除正常组外,应用脑立体定位仪定位海马组织注射β样淀粉蛋白(Aβ1-42)造模,并给予小柴胡汤化裁方高中低剂量(40 g·kg-1、20 g·kg-1、10 g·kg-1)进行干预,西药组用尼莫地平(15 mg·kg-1)对照。用免疫组化法观察细胞炎性因子IL-1β、IL-6及磷酸化p-p38 MARK信号分子表达情况,用Western blotting方法检测NF-κB蛋白表达。结果与正常组对比,模型组的海马区阳性细胞最多,细胞排列松散,IL-1β、IL-6及p-p38MARK表达量均高于正常组;各中药组和西药组对比模型组均有不同程度的减轻,其中以中药高剂量组最为显著。模型组海马NF-κB蛋白明显表达量增加,中药治疗组及西药组干预后,NF-κB蛋白表达水平下降,以中药高剂量组表现较为突出,与正常组相对接近。讨论经方小柴胡汤化裁方能并通过下调IL-1β、IL-6及p-p38MARK、NF-κB表达水平水平,表明小柴胡汤化裁方可能通过p38MAPK/NF-κB通路抑制Aβ对神经细胞的损伤,进而对AD模型小鼠发挥脑保护作用。

Objective To observe the effect of Xiaochaihu Decoction on the expression levels of inflammatory factors IL-1β, IL-6, p-p38 MARK and NF-κB in hippocampus of Alzheimer’s disease mice and explore its mechanism. Methods Thirty SPF mice were randomly divided into six groups. In addition to the normal group, the hippocampus tissue was injected with beta-amyloid protein(Aβ 1-42) by stereotaxic brain locator, and the high, middle and low dose of Xiaochaihu Decoction(40 g/kg, 20 g/kg, 10 g/kg) were given to intervene. The western medicine group was given nimodipine(15 mg/kg). The expressions of IL-1β, IL-6 and phosphorylated p-p38 MARK signal molecules were observed by immunohistochemistry, and the expression of NF-κB protein was detected by Western blotting. Result Compared with the normal group, the hippocampus of the model group had the most positive cells, the cells arranged loosely, and the expressions of IL-1β, IL-6 and p-p38 were higher than those of the normal group. Compared with those of the model group, the expressions of IL-1β, IL-6 and p-p38 MARK in the model group were alleviated to varying degrees, especially in the high-dose group of traditional Chinese medicine. The expression of NF-κB protein in hippocampus of the model group increased obviously. After the intervention of traditional Chinese medicine treatment and Western medicine, the expression level of NF-κB protein decreased, especially in the high dose group of traditional Chinese medicine, which was close to that of the normal group. Conclusion Xiaochaihu Decoction can reduce the expression levels of IL-1β, IL-6, p-p38 MARK and NF-κB, indicating that Xiaochaihu Decoction may inhibit the neuronal damage caused by Aβ through the p38 MAPK/NF-κB pathway, and thus exerting brain protective effects on Alzheimer’s disease model mice.