枸杞多糖对大鼠角膜缘干细胞体外增殖的影响研究  

Research of effects of lyciumbarbarum polysaccharides on proliferation of the limbal stemcells in vitro

作　　者：梁铮[1] 利嘉琦[2] 吕玉华 蔡志梁 晁艳[1] 周厚全[1] 卢燕君[1] Liang Zheng;Li Jiaqi;Lv Yuhua;Cai Zhiliang;Chao Yan;Zhou Houquan(Blood Transfusion Department,Guangdong Provincial Hospital of Traditional Chinese Medicine,Guangzhou 510120,China;Guangdong Center for Clinical Laboratory,Guangdong Provincial People's Hospital,Guangzhou 510080,China;The Second Clinical School of Guangzhou University of Chinese Medicine,Guangzhou 510405,China)

机构地区：[1]广东省中医院输血科,广州510120 [2]广东省人民医院广东省临床检验中心,广州510080 [3]广州中医药大学第二临床医学院,广州510405

出　　处：《中华临床实验室管理电子杂志》2020年第4期246-250,共5页Chinese Journal of Clinical Laboratory Management(Electronic Edition）

基　　金：广东省中医药局科研项目(20181112);广东省中医药局科研项目(20202080)。

摘　　要：目的通过体外分离和培养大鼠角膜缘干细胞(Limbal Stem Cells,LSCs),观察枸杞多糖(Lyciumbarbarum Polysaccharide,LBP)对LSCs体外增殖的影响,为中药体外培养LSCs的研究提供实验资料。方法体外分离和培养LSCs,显微镜观察LSCs的细胞形态特征,通过免疫荧光检测转录因子p63初步鉴定大鼠LSCs;分对照组和枸杞多糖处理组,每组20例,免疫印迹检测增殖细胞核抗原(Proliferating Cell Nuclear Antigen,PCNA),初步探讨枸杞多糖对LSCs的增殖影响。结果镜下观察LSCs体积小,呈多边形或椭圆形,胞核较大,核质比例大,免疫荧光结果显示体外分离的LSCs可表达p63(阳性率为76.41±1.66%);枸杞多糖处理后LSCs的PCNA蛋白的表达水平高于对照组,并且随着枸杞多糖浓度的增加,PCNA蛋白的表达升高(P<0.05)。结论枸杞多糖对LSCs的增殖和生长具有促进和维持的作用。Objective To observe the effects of Lyciumbarbarum polysaccharide(LBP)on the proliferation of limbal stem cells(LSCs)by isolation and culture of LSCs of rat in vitro,and to provide experimental data for the study of LSCs cultured in traditional Chinese medicine in vitro.Methods LSCs were isolated and subcultured in vitro,which were preliminarily identified by the characteristics of cells morphology with microscope and the detection of transcription factor p63 with immunofluorescence.Assess the impact of LBP on LSCs proliferation.Control group and LBP group with 20 cases in each group.The expression of proliferating cell nuclear antigen(PCNA)was detected by Western blot.Results LSCs observed under the microscope were small in size,polygonal or elliptical,with large nuclei and a large proportion of nucleus/cytoplasm.Immunofluorescence showed that isolation and culture of LSCs of rat in vitro could express p63 with positive rate 76.41±1.66%;the expression of PCNA in the LBP group was significantly higher than that in the control group(P<0.05).Conclusion LSCs were isolated and cultured in vitro preliminarily,and LBP can promote and maintain the proliferation and growth of LSCs in vitro.

关 键 词：角膜缘干细胞 枸杞多糖 体外培养 P63 PCNA 

分 类 号：R285.5[医药卫生—中药学]