干眼寒燥证大鼠模型建立与验证  

Establishment and Validation of Dry Eye Rat Model with Cold Dryness Syndrome

作　　者：陈立浩 陈雄[1] 彭清华[1] 姚小磊[3,4] 彭俊 汤钰[1] CHEN Lihao;CHEN Xiong;PENG Qinghua;YAO Xiaolei;PENG Jun;TANG Yu(Hunan University of Chinese Medicine,Changsha 410208,China;Hebei Eye Hospital,Xingtai 054001,China;The First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Hunan Provincial Key Laboratory for Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Chinese Medicine,Changsha 410208,China;Hunan Provincial Engineering and Technological Research Center for Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Chinese Medicine and Protecting Visual Function,Changsha 410208,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]河北省眼科医院,河北邢台054001 [3]湖南中医药大学第一附属医院,湖南长沙410007 [4]中医药防治眼耳鼻咽喉疾病湖南省重点实验室,湖南长沙410208 [5]湖南省中医药防治眼耳鼻咽喉疾病与视功能保护工程技术研究中心,湖南长沙410208

出　　处：《中国中医药信息杂志》2022年第12期76-81,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(30772824、82174443);湖南省自然科学基金面上项目(2021JJ3027);中医药防治眼耳鼻喉疾病湖南省重点实验室建设项目(2017TP1018);湖南省中医药防治眼耳鼻咽喉疾病与视功能保护工程技术研究中心开放基金(2018YGC02);湖南省教育厅科学研究重点项目(2020年);湖南省中医药科研计划重点项目(2021年);国家中医药管理局中医眼科学重点学科建设项目(ZK1801YK015);长沙市科技计划项目(K1501014-31、kc1704005);湖南省研究生科研创新项目(CX20200806);湖南中医药大学中医学国内一流建设学科(2018年)。

摘　　要：目的建立干眼寒燥证大鼠模型,通过检测中药干预后大鼠泪腺、睑板腺变化,验证模型是否成功。方法利用低温低湿小动物逆境实验箱建立干眼寒燥证大鼠模型,将大鼠随机分为空白组、模型组、中药组和灸法组,每组6只。在造模第1、7、21日分别检测大鼠泪液分泌量和泪膜破裂时间,记录大鼠造模前后角膜荧光素染色及特征性舌象。第22日起,中药组和灸法组分别用对证中药、灸法对大鼠进行干预,第28日行泪液分泌量及泪膜破裂时间检测后处死大鼠,取泪腺、睑板腺组织,HE染色观察病理变化,TUNEL染色检测细胞凋亡。结果造模后大鼠泪液分泌量减少(P<0.05),泪膜破裂时间缩短(P<0.05),角膜混浊,有不同程度的点状、片状侵损;舌体出现干燥、萎缩等燥证舌象改变。中药组大鼠泪液分泌量增加(P<0.05),泪膜破裂时间有所延长(P>0.05)。HE染色结果显示,模型组大鼠睑板腺组织腺泡紊乱,泪腺组织有大量炎性细胞浸润,中药组和灸法组大鼠睑板腺组织腺泡紊乱有所改善,泪腺组织炎性细胞浸润明显减少。TUNEL染色结果显示,与空白组比较,模型组大鼠睑板腺和泪腺组织细胞凋亡率显著增加(P<0.01);与模型组比较,中药组和灸法组大鼠睑板腺和泪腺组织细胞凋亡率显著减少(P<0.05)。结论本实验成功建立了干眼寒燥证大鼠模型。中药对证治疗可以减少干眼寒燥证模型大鼠睑板腺、泪腺组织炎性细胞浸润及细胞凋亡,增加泪液分泌,并在一定程度上改善泪膜稳定性。Objective To establish a rat model of dry eye with cold dryness syndrome;To validate the modeling by detecting the changes of lacrimal gland and meibomian gland in rats after the intervention of TCM.Methods The dry eye with cold dryness syndrome rat model was established in the cold and dry environment using the highprecision artificial climate box.The rats were randomly divided into blank group,model group,TCM group and moxibustion group,with 6 rats in each group.On the 1st,7th,and 21st days of modeling,the tear secretion and tear film break-up time of rats were detected,respectively.Corneal fluorescein staining and characteristic tongue images of rats were recorded before and after the intervention.From the 22nd day,the rats in TCM group and moxibustion group were intervened with TCM and moxibustion,respectively,and were killed after the detection of tear secretion and tear film break-up time on the 28th day.The lacrimal gland and meibomian gland of rats were taken,the pathological changes were observed by HE staining and the apoptosis was detected by TUNEL staining.Results After modeling,the amount of tear secretion decreased(P<0.05),and the tear film break-up time was shortened(P<0.05),corneal opacity with varying degrees of punctate and flake invasion;the tongue body appeared dry,atrophy and other tongue changes of dryness syndrome.In TCM group,the amount of tear secretion significantly increased(P<0.05),the tear film break-up time was prolonged(P>0.05).The results of HE staining showed that the rats in model group had disordered meibomian gland acini and a large number of inflammatory cells infiltration in lacrimal gland,the disorder of meibomian gland acini in TCM group and moxibustion group improved,and the inflammatory cells infiltration in lacrimal gland was significantly reduced.TUNEL staining showed that compared with the blank group,the apoptosis rate in meibomian gland and lacrimal gland tissue of rats in the model group increased significantly(P<0.01);Compared with the model group,the apoptosis rat

关 键 词：干眼 寒燥证 泪腺 睑板腺 炎症 凋亡 大鼠 

分 类 号：R285.5[医药卫生—中药学]