黄连解毒汤对APP/PS1双转基因AD小鼠脑组织IL-6I、L-1β水平及SP、NFTs数量的干预作用

Study on influence of Huanglianjiedu decoction on contents of IL-6 and IL-1β and numbers of SP and NFTs in brain of APP/PS1 double transgenic AD mice

目的通过观察黄连解毒汤对APP/PS1双转基因阿尔茨海默病(AD)小鼠脑组织炎性细胞因子白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)水平及海马区老年斑(SP)、神经原纤维缠结(NFTs)数量的影响,探讨黄连解毒汤治疗AD的可能机制,以及其与西药盐酸多奈哌齐的疗效比较。方法 3月龄雄性SPF级APP/PS1双转基因AD小鼠分别经盐酸多奈哌齐或黄连解毒汤大、中、小剂量灌胃7个月后,应用酶联免疫吸附法(ELISA)检测其脑组织中炎性细胞因子IL-6I、L-1β水平,免疫荧光染色法检测小鼠海马区SP及NFTs数量并观察神经细胞核病理学变化,并进行分析。结果与模型溶剂对照组比较,盐酸多奈哌齐组及中药各剂量组IL-6水平均显著降低、神经细胞核病变发生率(PNND)均显著减低(P<0.01),SP数量均减少(P<0.01、P<0.01、P<0.01、P<0.05),NFTs形成均减少(P<0.05)。与盐酸多奈哌齐组比较,中药大、中、小剂量组IL-6水平均降低(P<0.05、P<0.01、P<0.05),中药中剂量组SP数量减少(P<0.05)、NFTs形成减少(P<0.01),PNND减低(P<0.01)。中药大、中剂量组较小剂量组SP数量减少(均P<0.01);NFTs形成大剂量组较中剂量组增多(P<0.05),中剂量组较小剂量组减少(P<0.01);大剂量组较中、小剂量组PNND增高(P<0.01、P<0.05),中剂量组较小剂量组PNND减少(P<0.01)。中药各剂量组间IL-6水平比较、各实验组组间IL-1β水平比较无统计学差异(均P>0.05)。结论中药黄连解毒汤同盐酸多奈哌齐均具有神经细胞保护作用,但其疗效可能优于盐酸多奈哌齐。

Objective To observe the influence of Huanglianjiedu decoction （HJD） on contents of interleukin-6 （IL-6） and interleukin-1β （IL-1β） and numbers of senile plaques （SP） and neurofibrillary tangles （NFTs） in brain of APP/PS1 double transgenic Alzheimer disease （AD） mice. To discuss the possible mechanism of AD treated by HJD and to compare the effect between HJD and hydrochloride acid donepezil. Methods After 7-month gavage treatment by high, middle and low doses of HJD or hydrochloride acid donepezil respectively, enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of inflammatory cytokines IL-6 and IL-1β in APP/PS1 double transgenic AD mice brain, lmmunofluorescence staining was used to observe the numbers and morphology of SP and NFTs, and pathological changes of nerve cell nucleus in hippocampus area through fluorescence microscope. Results Compared with the control group, IL-6 levels and the percentage of denaturation of Neurons nucleus （PNND） reduced in the hydrochloride acid donepezil group and the HJD groups （P〈0. 01）; the number of SP reduced in the hydrochloride acid donepezil group and the high, middle and low dose HJD group （P%0.01, P d0.01, P〈0. 01, P〈0.05）; the formation of NFTs decreased in HJD groups and hydrochloric acid donepezil group （P〈 0.01, P 〈0.05）. Compared with the hydrochloric acid donepezil group, IL-6 levels decreased in the high, middle and small close HJD groups （P 〈 0.05, P 〈0.01, P 〈0.05）; the number of SP reduced in the middle HJD dose group （P 〈0.05）; the formation of NFTs reduced in the middle dose group （P%0.01） ; PNND reduced in the middle dose HJD group （P 〈0.01）. The number of SP was less in both the high and middle dosage HJD groups than in the small dose group （P 〈0.01） ; the formation of NFTs was more in the high and small dose groups than in the middle dose group （P 〈0.05, P 〈0.01） ; the PNND increased in the high dose HJD group compared with the middle and small dosage HJD groups （P 〈0.01, P〈0.05）, and the PNND reduced in the middle dosage HJD group compared with the small dose group （P 〈0.01）. There were no significant differences in IL-6 levels among the HJD groups and IL-1β levels among all the groups （P〉 0.05）. Conclusions The neuronal cells could be protected by treating with HJD or hydrocbloride acid donepezil. It seems that HJD was more effective than the hydrochloride acid donepezil in treating AD.