活血散结方含药血浆对视网膜色素上皮细胞毒性作用的研究
|
摘要
目的制备活血散结方含药血浆,并研究其不同浓度对视网膜色素上皮(retinal pigment epithelium,RPE)细胞的毒性作用。方法以成年健康家兔为对象,制备活血散结方含药血浆。将提取的兔原代RPE细胞分9个组,分别为:空白对照组、正常血浆组、5%含药血浆组、10%含药血浆组、20%含药血浆组、40%含药血浆组、60%含药血浆组、80%含药血浆组、100%含药血浆组,予以相对应浓度的含药血浆干预,CCK8法检测含药血浆对兔RPE细胞毒作用。结果活血散结方各浓度含药血浆对RPE细胞有抑制作用,其中5%～20%含药血浆对RPE细胞有较弱的抑制作用,但与正常血浆比较,抑制率差异无统计学意义(P>0.05);而40%～100%含药血浆对RPE细胞生长有明显的抑制作用,与正常血浆比较,抑制率差异有统计学意义(P<0.05)。结论活血散结方含药血浆对RPE细胞有一定的抑制作用,抑制程度与含药血浆浓度成量效关系,可以考虑选择5%～20%含药血浆浓度作为后续实验研究的干预浓度。
Objective To prepare Huoxue Sanjie Decoction-containing plasma, and to study the toxic effect of different concentrations of the plasma on retinal pigment epithelium(RPE) cells. Methods Adult healthy rabbits were used to prepare Huoxue Sanjie Decoction-containing plasma. Rabbit primary cells were divided into nine groups: blank control group, normal plasma group, 5% drug-containing plasma group, 10% drug-containing plasma group, 20% drug-containing plasma group, 40%drug-containing plasma group, 60% drug-containing plasma group, 80% drug-containing plasma group, and 100% drug-containing plasma group. These groups were treated with respective concentrations of drug-containing plasma. CCK-8 assay was used to examine their cytotoxic effect on rabbit RPE cells. Results All concentrations of Huoxue Sanjie Decoction-containing plasma had an inhibitory effect on RPE cells. The 5%, 10%, and 20% drug-containing plasma had a weak inhibitory effect on RPE cells,which was not significantly different from the inhibitory effect of normal plasma(P>0.05). However, the 40%, 60%, 80%, and 100%drug-containing plasma had an obvious inhibitory effect on RPE cell growth, and the inhibition rates were significantly from that of the normal plasma group(P<0.05). Conclusion Huoxue Sanjie Decoction-containing plasma has a certain inhibitory effect on RPE cells, which is correlated with the concentration of drug-containing plasma. The 5%-10% concentration of drug-containing plasma can be considered as the intervention concentration in the following experimental studies.
Objective To prepare Huoxue Sanjie Decoction-containing plasma, and to study the toxic effect of different concentrations of the plasma on retinal pigment epithelium(RPE) cells. Methods Adult healthy rabbits were used to prepare Huoxue Sanjie Decoction-containing plasma. Rabbit primary cells were divided into nine groups: blank control group, normal plasma group, 5% drug-containing plasma group, 10% drug-containing plasma group, 20% drug-containing plasma group, 40%drug-containing plasma group, 60% drug-containing plasma group, 80% drug-containing plasma group, and 100% drug-containing plasma group. These groups were treated with respective concentrations of drug-containing plasma. CCK-8 assay was used to examine their cytotoxic effect on rabbit RPE cells. Results All concentrations of Huoxue Sanjie Decoction-containing plasma had an inhibitory effect on RPE cells. The 5%, 10%, and 20% drug-containing plasma had a weak inhibitory effect on RPE cells,which was not significantly different from the inhibitory effect of normal plasma(P>0.05). However, the 40%, 60%, 80%, and 100%drug-containing plasma had an obvious inhibitory effect on RPE cell growth, and the inhibition rates were significantly from that of the normal plasma group(P<0.05). Conclusion Huoxue Sanjie Decoction-containing plasma has a certain inhibitory effect on RPE cells, which is correlated with the concentration of drug-containing plasma. The 5%-10% concentration of drug-containing plasma can be considered as the intervention concentration in the following experimental studies.
引文
[1]薛晓辉.VEGF和bFGF在增生性玻璃体视网膜病变患者玻璃体中的表达及意义[J].陕西医学杂志,2016,45(6):707-708,719.
[2]董尼娜.干扰素-γ联合曲安奈德防治兔增生性玻璃体视网膜病变的实验研究[D].郑州:郑州大学,2014.
[3]陶勇,姜燕荣,高新晓.外伤性视网膜脱离眼发生严重增生性玻璃体视网膜病变的危险因素研究[J].眼科新进展,2009,29(8):594-596.
[4]孙蕾.靶向PDGF-A RNA干涉抑制增生性玻璃体视网膜病变中RPE细胞增殖的实验研究[D].长春:吉林大学,2008.
[5]李兰根,伟伟,张玉凤,等.视网膜色素上皮细胞氧化应激过程中SirT1/STAT3的相互作用[J].眼科新进展,2017,37(12):1-3.
[6]路永珩,郑轶.增生性玻璃体视网膜病变发病机制与治疗的研究进展[J].医学综述,2018,24(13):2548-2552.
[7]焦明菲,颜华.增生性玻璃体视网膜病变的药物治疗[J].眼科研究,2010,28(4):381-384.
[8]陈吉,彭清华,邢雁飞,等.散血明目片对外伤性增生性玻璃体视网膜病变兔眼玻璃体结缔组织生长因子的影响[J].中国中医眼科杂志,2006,16(4):224-227.
[9]张亚妮.散结明目片对兔视网膜保护及抗增殖作用的实验研究[D].乌鲁木齐:新疆医科大学,2010.
[10]彭清华,刘娉,彭俊,等.益气养阴活血利水中药复方对兔视网膜脱离后视网膜组织中ATP含量的影响[J].湖南中医药大学学报,2016,36(7):28-30.
[11]肖华,范钦华.中药干预对防治增生性玻璃体视网膜病变的基础研究[J].四川中医,2013,31(11):156-158.
[12]王文丽.海昆化瘀片调控RPE诱导的增生性玻璃体视网膜病变TGF-β表达的研究[D].成都:成都中医药大学:2013.
[13]XU G X,YANG J,SUN T S,et al.Primary culture of human retinal pigment epithelium in vitro[J].International Journal of Ophthalmology,2004,4(1):12-15.
[14]胡健艳,陈雪,陈辉,等.兔视网膜色素上皮细胞培养和鉴定[J].南通大学学报,2005,25(3):171-173.
[15]彭清华,彭俊.暴盲病名沿革及分化[J].中华中医药学刊,2010,28(9):1812-1813.
[16]王文丽,叶河江,张晓婷,等.从眼内癥瘕论治增生性玻璃体视网膜病变[J].四川中医,2013,31(4):33-34.
[17]吴要华.化瘀散结片对DispaseI诱导的实验性增生性玻璃体视网膜病变müller细胞的影响[D].成都:成都中医药大学,2013.
[18]付美林,彭清华,陈吉,等.活血利水法对兔外伤性PVR增殖膜上EGFmRNA表达的影响[J].国际眼科杂志,2012,12(1):25-29.
[19]余肖.三七对防治外伤性增生性玻璃体视网膜病变的实验研究[D].南京:南京中医药大学,2014.
[20]林泽苗,钟佳贤,李青南.中药血清药理学和血浆药理学应用比较研究[J].亚太传统医药,2016,12(12):62-64.
[21]刘林.中药含药血浆与血清有效成分比较及血浆药理学方法研究[D].长沙:湖南中医药大学,2016.
[2]董尼娜.干扰素-γ联合曲安奈德防治兔增生性玻璃体视网膜病变的实验研究[D].郑州:郑州大学,2014.
[3]陶勇,姜燕荣,高新晓.外伤性视网膜脱离眼发生严重增生性玻璃体视网膜病变的危险因素研究[J].眼科新进展,2009,29(8):594-596.
[4]孙蕾.靶向PDGF-A RNA干涉抑制增生性玻璃体视网膜病变中RPE细胞增殖的实验研究[D].长春:吉林大学,2008.
[5]李兰根,伟伟,张玉凤,等.视网膜色素上皮细胞氧化应激过程中SirT1/STAT3的相互作用[J].眼科新进展,2017,37(12):1-3.
[6]路永珩,郑轶.增生性玻璃体视网膜病变发病机制与治疗的研究进展[J].医学综述,2018,24(13):2548-2552.
[7]焦明菲,颜华.增生性玻璃体视网膜病变的药物治疗[J].眼科研究,2010,28(4):381-384.
[8]陈吉,彭清华,邢雁飞,等.散血明目片对外伤性增生性玻璃体视网膜病变兔眼玻璃体结缔组织生长因子的影响[J].中国中医眼科杂志,2006,16(4):224-227.
[9]张亚妮.散结明目片对兔视网膜保护及抗增殖作用的实验研究[D].乌鲁木齐:新疆医科大学,2010.
[10]彭清华,刘娉,彭俊,等.益气养阴活血利水中药复方对兔视网膜脱离后视网膜组织中ATP含量的影响[J].湖南中医药大学学报,2016,36(7):28-30.
[11]肖华,范钦华.中药干预对防治增生性玻璃体视网膜病变的基础研究[J].四川中医,2013,31(11):156-158.
[12]王文丽.海昆化瘀片调控RPE诱导的增生性玻璃体视网膜病变TGF-β表达的研究[D].成都:成都中医药大学:2013.
[13]XU G X,YANG J,SUN T S,et al.Primary culture of human retinal pigment epithelium in vitro[J].International Journal of Ophthalmology,2004,4(1):12-15.
[14]胡健艳,陈雪,陈辉,等.兔视网膜色素上皮细胞培养和鉴定[J].南通大学学报,2005,25(3):171-173.
[15]彭清华,彭俊.暴盲病名沿革及分化[J].中华中医药学刊,2010,28(9):1812-1813.
[16]王文丽,叶河江,张晓婷,等.从眼内癥瘕论治增生性玻璃体视网膜病变[J].四川中医,2013,31(4):33-34.
[17]吴要华.化瘀散结片对DispaseI诱导的实验性增生性玻璃体视网膜病变müller细胞的影响[D].成都:成都中医药大学,2013.
[18]付美林,彭清华,陈吉,等.活血利水法对兔外伤性PVR增殖膜上EGFmRNA表达的影响[J].国际眼科杂志,2012,12(1):25-29.
[19]余肖.三七对防治外伤性增生性玻璃体视网膜病变的实验研究[D].南京:南京中医药大学,2014.
[20]林泽苗,钟佳贤,李青南.中药血清药理学和血浆药理学应用比较研究[J].亚太传统医药,2016,12(12):62-64.
[21]刘林.中药含药血浆与血清有效成分比较及血浆药理学方法研究[D].长沙:湖南中医药大学,2016.