龙牡安神方通过调节内侧前额叶皮质小胶质细胞极化改善CRS小鼠的焦虑行为  

Longmu Anshen prescription alleviates anxiety behavior in CRS mice by regulating polarization of microglia in medial prefrontal cortex

作　　者：彭晓明 丁晓华[2,3] 张震文 赵建军[2] 唐珂 PENG Xiaoming;DING Xiaohua;ZHANG Zhenwen;ZHAO Jianjun;TANG Ke(Affiliated Hospital of Gansu University of Traditional Chinese Medicine,Lanzhou 730000,China;Gansu University of Traditional Chinese Medicine,Lanzhou 730000,China;Gansu Provincial Maternity and Child Care Hospital,Lanzhou 730050,China)

机构地区：[1]甘肃中医药大学附属医院,兰州730000 [2]甘肃中医药大学,兰州730000 [3]甘肃省妇幼保健院,兰州730050

出　　处：《中国免疫学杂志》2024年第11期2316-2321,2329,共7页Chinese Journal of Immunology

基　　金：甘肃省自然科学基金(20JR10RA338);2022年甘肃中医药大学研究生创新创业基金(2022CX42)。

摘　　要：目的:探讨龙牡安神方对慢性束缚应激(CRS)小鼠内侧前额叶皮层(mPFC)小胶质细胞(MG)极化及焦虑行为的影响。方法:48只健康雄性昆明小鼠随机分为正常组、盐酸帕罗西汀组和龙牡安神方高、中、低剂量组。除正常组外,其余各组采用CRS构建焦虑模型,持续束缚35 d,从第15天开始,每日CRS 30 min后,正常组和模型给予蒸馏水灌胃,盐酸帕罗西汀组给予盐酸帕罗西汀(2.6 mg/kg)灌胃,中药组给予龙牡安神方高、中、低剂量(4.16、2.08、1.04 g/kg)灌胃,给药时间为21 d。通过明暗箱实验和高架十字迷宫评价小鼠焦虑行为的变化;ELISA检测小鼠mPFC组织匀浆中IL-1β、IL-6、TNF-α及IL-10水平;Western blot检测小鼠mPFC组织中诱导型一氧化氮合酶(iNOS)和精氨酸酶1(Arg-1)蛋白表达;RT-PCR检测小鼠mPFC组织中离子钙接头蛋白分子-1(Iba-1)、iNOS和Arg-1 mRNA的表达;免疫组化(IHC)检测小鼠mPFC组织中Iba-1蛋白的表达。结果:与正常组比较,模型组小鼠明箱停留时间、明暗箱穿梭次数、进入开臂次数百分比(OE%)及开臂停留时间百分比(OT%)均显著减少(P<0.01);模型组小鼠mPFC组织中Iba-1、iNOS蛋白及mRNA表达显著升高(P<0.01),IL-1β、IL-6和TNF-α水平显著升高(P<0.01),而Arg-1的蛋白及mRNA表达显著降低(P<0.01),IL-10水平显著降低(P<0.01)。与模型组比较,盐酸帕罗西汀组及龙牡安神方高、中剂量组小鼠明暗箱停留时间、明暗箱穿梭次数、OE%及OT%均显著增加(P<0.01);mPFC组织中Iba-1、iNOS蛋白及mRNA表达明显降低(P<0.05,P<0.01),IL-1β、IL-6和TNF-α水平显著降低(P<0.01),而Arg-1的蛋白及mRNA表达显著升高(P<0.01),IL-10水平显著升高(P<0.01)。结论:龙牡安神方能够抑制CRS小鼠mPFC组织中MG的激活,并可能通过调节M1/M2极化表型减轻神经炎症,增加抗炎细胞因子的水平,从而改善CRS诱导的焦虑行为。Objective:To investigate the effects of Longmu Anshen prescription on microglia(MG)polarization in the medial prefrontal cortex(mPFC)and anxiety behavior of chronic restraint stress(CRS)mice.Methods:Forty-eight healthy male KM mice were randomly divided into normal group,paroxetine hydrochloride group and Longmu Anshen prescription high,middle and low dose groups.From the 15th day,after 30 minutes of CRS every day,the normal group and the model were given distilled water by gavage,the paroxetine hydrochloride group was given paroxetine hydrochloride(2.6 mg/kg)by gavage,and the traditional Chinese medicine group was given Longmu Anshen prescription at high,medium and low doses(4.16,2.08,1.04 g/kg)by gavage for 21 days.The changes of anxiety behavior in mice were evaluated by light-dark boxes experiment and elevated plus maze;ELISA was used to detect IL-1β,IL-6,TNF-αand IL-10 levels;protein expression of inducible nitric oxide synthase(iNOS)and arginase-1(Arg-1)in mice mPFC tissue were detected by Western blot;mRNA expression of ionized calcium binding adaptor molecule-1(Iba-1),iNOS and Arg-1 in mice mPFC tissue were detected by RT-PCR;the protein expression of Iba-1 in mice hippocampal tissue was detected by immuno-histochemistry(IHC).Results:Compared with normal group,the residence time in the light boxes,the number of times of shuttle between the light-dark boxes,the percentage of times of entering the open arm(OE%)and the percentage of time spent in the open arm(OT%)of mice in model group were significantly reduced(P<0.01);moreover,in the mPFC tissue of the model group mice,the protein and mRNA expressions of Iba-1 and iNOS were significantly increased(P<0.01),and the levels of IL-1β,IL-6 and TNF-αwere significantly increased(P<0.01),while the protein and mRNA expressions of Arg-1 were significantly decreased(P<0.01),and the level of IL-10 was significantly decreased(P<0.01).Compared with model group,the residence time in the light boxes,the number of shuttles between the light-dark boxes,the OE%and OT%of m

关 键 词：龙牡安神方 慢性束缚应激(CRS) 焦虑 小胶质细胞 极化 

分 类 号：R285.5[医药卫生—中药学]