左旋紫草素对小鼠变应性鼻炎抑制作用研究  

Inhibitory effect of L-shikonin on allergic rhinitis in mice

作　　者：陈武兵 王艺龙 应永杰 CHEN Wubing;WANG Yilong;YING Yongjie(Department of Otolaryngology,Municipal Hospital Affiliated to Taizhou University,Taizhou Municipal Hospital,Taizhou 318000,China)

机构地区：[1]台州学院附属市立医院,台州市立医院耳鼻喉科,浙江台州318000

出　　处：《空军军医大学学报》2024年第6期699-704,共6页Journal of Air Force Medical University

基　　金：浙江省医药卫生科技计划项目(2023KY1325);浙江省中医药科技计划项目(2024ZL1246)。

摘　　要：目的探讨左旋紫草素对变应性鼻炎(AR)小鼠治疗效应及其相关抑制机制。方法采用卵清蛋白(OVA)致敏建立小鼠AR模型,按随机数字表法将25只C57BL/6小鼠随机分为正常组、AR模型组、左旋紫草素低剂量和高剂量组,以及标准对照组,每组5只。除正常组外,其他各组完成AR模型构建:在第1、5、9、12日使用OVA+氢氧化铝凝胶混合制剂进行致敏,第13日开始连续7 d,使用OVA每日进行一次激发。在造模第28日起,左旋紫草素组小鼠灌胃2、8 mg/(kg·d)左旋紫草素。标准对照组小鼠灌胃给予地塞米松2.5 mg/(kg·d),正常组和AR模型组动物灌胃等剂量的二甲基亚砜,共灌胃4周,末次灌胃30 min内观察各组小鼠鼻部症状并予以评分。采用流式细胞术检测小鼠外周血辅助性T细胞(Th)1和Th2细胞亚群表达水平;ELISA检测小鼠鼻腔灌洗液白细胞介素4(IL-4)、免疫球蛋白E(IgE)、γ干扰素(IFN-γ)水平;HE染色检测鼻黏膜病理情况。qRT-PCR法检测鼻黏膜组织中T盒子转录因子(T-bet)、GATA结合蛋白3(GATA3)、信号传导子和转录活化子1(STAT1)、STAT6 mRNA表达。结果给药4周后,与正常组相比,AR模型组的行为学评分、Th1细胞比例、IL-4和IgE水平、GATA3和STAT6 mRNA表达显著增加(P<0.05),Th2细胞比例、IFN-γ水平、T-bet和STAT1 mRNA表达显著降低(P<0.05)。与AR模型组相比,左旋紫草素低剂量组动物的行为学评分,Th1和Th2细胞比例,IL-4、IgE和IFN-γ水平,T-bet mRNA表达几乎无改变(P>0.05),STAT1 mRNA表达显著提高(P<0.05),而GATA3和STAT6 mRNA表达显著降低(P<0.05);与AR模型组相比,左旋紫草素高剂量组动物行为学评分、Th1细胞比例、IL-4和IgE水平、GATA3和STAT6 mRNA表达显著下降(P<0.05),Th2细胞比例、IFN-γ水平、T-bet和STAT1 mRNA表达显著增加(P<0.05)。与左旋紫草素低剂量组相比,左旋紫草素高剂量组外周血中Th1细胞比例、IL-4和IgE水平、GATA3和STAT6 mRNA表达显著下降(P<0.05),Th2�Objective To investigate the therapeutic effect of L-shikonin on mice with allergic rhinitis(AR)and its related inhibitory mechanism.Methods Ovalbumin(OVA)-sensitized AR mouse model was established.According to random number table method,25 C57BL/6 mice were randomly divided into normal group,AR model group,low-dose and high-dose L-shikonin groups,and standard control group,with 5 mice in each group.In addition to the normal group,the other groups completed AR model construction:OVA+aluminum hydroxide gel mixed preparation was used for sensitization on the 1st,5th,9th and 12th day,and OVA was used for stimulation once a day for seven consecutive days starting from the 13th day.Starting from the 28th day of modeling,the mice in the L-shikonin group were administered with 2 and 8 mg/(kg·Fd)L-shikonin by gavage.The mice in the standard control group were given 2.5 mg/(kg·Fd)of dexamethasone by gavage,while the normal group and AR model group were given an equal dose of dimethyl sulfoxide by gavage for a total of 4 weeks.Within 30 min of the last gavage,nasal symptoms of each group of mice were observed and scored.Flow cytometry was used to detect the expression levels of T helper(Th)1 and Th2 cell subsets in peripheral blood of mice.ELISA was used to detect the levels of interleukin-4(IL-4),immunoglobulin E(IgE),andγ-interferon(IFN-γ)in the nasal lavage fluid of mice.HE staining was used to detect pathological changes in the nasal mucosa.qRT-PCR was used to detect the mRNA expression of T-box transcription factor(T-bet),GATA binding protein 3(GATA3),signal transducer and activator of transcription 1(STAT1),and STAT6 in nasal mucosal tissue.Results After 4 weeks of administration,compared with normal group,behavioral scores,Th1 cell proportion,IL-4 and IgE levels,and GATA3 and STAT6 mRNA expressions were significantly increased in AR model group(P<0.05),while Th2 cell proportion,IFN-γlevel,and T-bet and STAT1 mRNA expressions were significantly decreased in AR model group(P<0.05).Compared with AR model group,beh

关 键 词：左旋紫草素 变应性鼻炎 C57BL/6小鼠模型 TH1/TH2细胞因子 

分 类 号：R285.5[医药卫生—中药学]