“改良三甲散”含药脑脊液对Aβ_(25-35)诱导海马神经神经元细胞损伤IL-1α、IL-1β和IL-6的影响

Effect of Cerebrospinal Fluid with Drug-containing Modified San-Jia-San Decoction on IL-1α, IL-1βand IL-6 of Hippocampal Neurons Model Injury Induced by Aβ_(25-35)

目的：探讨改良三甲散对β淀粉样蛋白（Aβ25-35）所致海马神经元细胞损伤时细胞炎性因子IL-1α、IL-1β和IL-6的影响。方法：将原代培养的神经元细胞分为空白组、模型组、石杉碱甲组和改良三甲散低、中、高剂量组。选择性培养7~10天后吸去培养液，分别加入空白培养液、正常脑脊液（生理盐水组）、石杉碱甲脑脊液和中药脑脊液低、中和高剂量，加培养液补至等量，37℃孵育2 h。然后除空白组之外的各组加入经老化处理的Aβ25-35（终浓度为5μmol·L^-1），建立AD细胞模型。空白组加入等量的培养液，继续孵育24 h后收集细胞上清液。采用酶联免疫吸附测定法（ELISA）检测IL-1α、IL-1β和IL-6的含量。结果：改良三甲散能降低细胞上清液中IL-1α、IL-1β和IL-6的含量。结论：改良三甲散脑脊液能有效抑制细胞炎性因子IL-1α、IL-1β和IL-6的释放，具有一定的抗炎效应，从而保护海马神经元细胞。

This study was aimed to investigate effect on the expression of cell inflammatory factor IL-1α, IL-1β and IL-6 of modified San-Jia-San （SJS） decoction on cultured hippocampal neurons injury induced by β-amyloid 25-35 protein （Aβ25-35）. Primary cultured hippocampal neurons were divided into the normal group, model group, huperzine A group, low-dose SJS group, middle-dose SJS group, and high-dose SJS group. After selective culture for 7~10 days and the absorption of culture fluid, the blank culture fluid, normal cerebrospinal fluid （normal saline group）, huperzine A cerebrospinal fluid, and low-, middle- and high-dose SJS cerebrospinal fluid were added, respectively. The culture fluid was added up to an equivalent medium. The incubation was 2 h under the temperature of 37℃. Ex-cept the normal group, Aβ25-35 dealt with aging （the final concentration was 5 μmol/L） was added to other groups to establish AD cell model. An equivalent amount of culture fluid was added to the normal group. After 24 h of incuba-tion, the cell culture supernatant was collected. Enzyme-linked immunosorbent assay （ELISA） was used in the content detection of IL-1α, IL-1β and IL-6. The results showed that modified SJS decoction can decrease the content of IL-1α, IL-1β and IL-6 in the cell culture supernatant. It was concluded that the modified SJS decoction can effectively inhibit the expression of cell inflammatory factor IL-1α, IL-1β and IL-6. It had certain anti-inflammatory effect to protect hippocampal neurons.