不同剂量血小板源性生长因子-B及黄芪注射液对体外高糖培养大鼠视网膜毛细血管周细胞的影响  

Effect of different doses of Platelet-derived growth factor-B and astragalus injection on rat retinal microvascular pericytes cultured with high glucose in vitro

作　　者：谢俪君 宫晓红[2] 高颖[2] 周剑[2] 宋哲[2] 朱晓博 孙艳红[2] 郑敏[3] 达卓立 邵宝仪 XIE Lijun;GONG Xiaohong;GAO Ying;ZHOU Jian;SONG Zhe;ZHU Xiaobo;SUN Yanhong;ZHENG Min;DA Zhuoli;SHAO Baoyi(Mianyang Wanjiang Eye Hospital,Mianyang 621000,Sichuan,China;Dongfang Hospital,Beijing University of Traditional Chinese Medicine,Beijing 100078,China;Beijing University of Traditional Chinese Medicine,Beijing 100029,China;Shiping County Hospital of Traditional Chinese Medicine,Shiping 662200,Yunnan,China;School of Chinese Medicine,Hong Kong Baptist University,Hong Kong 999077,China)

机构地区：[1]绵阳万江眼科医院,四川绵阳621000 [2]北京中医药大学东方医院,北京100078 [3]北京中医药大学,北京100029 [4]云南省石屏县中医医院,云南石屏662200 [5]香港浸会大学中医药学院,香港999077

出　　处：《现代中西医结合杂志》2020年第27期2979-2982,3002,共5页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：北京中医药大学青年教师项目(2019-JYB-JS-096)。

摘　　要：目的观察不同剂量的血小板源性生长因子-B(PDGF-B)、不同剂量的黄芪注射液以及两者合用对体外高糖培养大鼠视网膜毛细血管周细胞(RMPs)生长的影响。方法取体外培养第3代大鼠RMPs,饥饿培养24 h后,分为空白组(5.5 mmol/L葡萄糖的低糖DMEM)、对照组(50 mmol/L葡萄糖的高糖DMEM)、不同浓度PDGF-B组(0.25 ng/mL、0.50 ng/mL、1.00 ng/mL+高糖DMEM)、不同浓度黄芪注射液组(0.2 mg/mL、2 mg/mL、20 mg/mL、200 mg/mL+高糖DMEM)和PDGF-B+不同浓度黄芪注射液组(1.00 ng/mL PDGF-B+0.2 mg/mL、2 mg/mL、20 mg/mL、200 mg/mL黄芪注射液+高糖DMEM),培养48 h后,采用CCK8法检测各组RMPs增殖情况,并用免疫荧光法鉴定增殖后的细胞。结果与对照组比较,不同浓度PDGF-B组,2 mg/mL、20 mg/mL、200 mg/mL黄芪注射液组及1.00 ng/mL PDGF-B+2 mg/mL、20 mg/mL、200 mg/mL黄芪注射液各组均能促进体外高糖培养的RMPs增殖,并在一定浓度内呈剂量-效应关系,差异均有统计学意义(P均<0.05);其中1.00 ng/mL PDGF-B组及200 mg/mL黄芪注射液组促增殖作用均优于1.00 ng/mL PDGF-B+200 mg/mL黄芪注射液组(P均<0.05)。加药培养后各组细胞表达周细胞标志物α-平滑肌肌动蛋白(α-SMA)及血小板源性生长因子受体-β(PDGFR-β),不表达血管性血友病因子(vWF),几乎不表达胶质纤维酸性蛋白(GFAP)。结论PDGF-B及黄芪注射液均对体外高糖培养大鼠RMPs的生长有促进作用,短期内未使RMPs发生变异,单用PDGF-B或黄芪注射液促生长作用优于两者合用。Objective It is to observe the effect of different doses of platelet-derived growth factor-B(PDGF-B),different doses of astragalus injection and their combination on the growth of rat retinal microvascular pericytes(RMPs)cultured with high glucose in vitro.Methods The third-generation rat RMPs were cultured in vitro after hungered 24h,which were divided into the blank group(cultured with low glucose DMEM,5.5 mmol/L),the control group(cultured with high glucose DMEM,50 mmol/L),different concentrations of PDGF-B groups(0.25,0.50,1.00 ng/mL+high glucose DMEM),different concentrations of astragalus injection groups(0.2,2,20,200 mg/mL+high glucose DMEM),and different concentrations of astragalus injection with PDGF-B groups(1.00 ng/mL PDGF-B+0.2,2,20,200 mg/mL astragalus injection+high glucose DMEM).After cultured for 48 h,RMPs proliferation in each group was detected by CCK8 method,and the cells were identified by immunofluorescence.Results Compared with the control group,PDGF-B(group 1,2,3),astragalus injection(group 2,3,4)and PDGF-B+astragalus injection(group 2,3,4)the proliferation of RMPs cultured in vitro with high glucose were all promoted in different concentrations of PDGF-B groups,2,20,200 mg/mL astragalus injection groups,and 1.0 ng/mL PDGF-B+2,20,200 mg/mL astragalus injection groups,which showed a dose-effect relationship within a certain concentration and had statistically significant difference(P<0.05).The effect on promoting proliferation of RMPs in 1ng/mL PDGF-B group and 200 mg/mL astragalus injection group was better than that in the 1ng/mL PDGF-B+200 mg/mL astragalus injection group(P<0.05).After adding the drugs to culture,the cells in each group expressed the pericytes markers α-smooth muscle actin(α-SMA)and Platelet-derived growth factor receptor-β(PDGFR-β),but did not express von Willebrand factor(vWF)and hardly expressed glial fibrillary acidic protein(GFAP).Conclusion PDGF-B and astragalus injection both could promote the growth of rat RMPs cultured with high glucose in vitro,without muta

关 键 词：糖尿病视网膜病变 视网膜毛细血管周细胞 血小板源性生长因子-B 黄芪注射液 

分 类 号：R774[医药卫生—眼科]