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山东省自然科学基金(Q2008C02)

作品数:2 被引量:4H指数:1
相关作者:杨玲玲王宜强宫华青王茜王瑶更多>>
相关机构:山东省眼科研究所更多>>
发文基金:山东省自然科学基金国家自然科学基金更多>>
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Comparison of the therapeutic effects of extracts from Spirulina platensis and amnion membrane on inflammation-associated corneal neovascularization被引量:4
2012年
AIM: To compare the therapeutic effects of polysaccharide extract from Spirulina platensis (PSP) and extract from amnion membrane CAME) on alkali burn-induced corneal neovascularization (CorNV). METHODS: PSP and AME were Extracted from dry powder of Spirulina platensis and human aminion membrane respectively. Murine CorNV was induced by applying 1N sodiumhydroxide (NaOH) solution directly on the mice corneas. PSP and AME extracts were administered topically on the corneas 4 times daily for 7 days. The therapy effects of PSP and AME extracts were evaluated daily using slit-lamp. At the end of the therapy, corneas were harvested for H&E staining, masson trichrome staining, immunohistochemical study, and semi-quantification reverse transcriptive PCR (RT-PCR) was utilized for measurement of inflammation-related molecules. RESULTS: Topical application of PSP extract had significant therapeutic effects on CorNV that could be shown in various assays of the corneas. Compared with AME Extract, PSP extract treatment was more effective in suppressing CorNV in berms of vessel length and levels of cluster of differentiation 31 (CD31) proteins or the angiogenesis related genes like vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP2) and matrix metalloproteinase-9 (MMP9). PSP also inhibited inflammation more markedly by more effectively inhibiting mononuclear and polymorphonuclear cells infiltration into the corneal stroma and reducing levels of stromal cell-derived factor-1 (SDF1), tumor necrosis factor-alpha (TNF alpha) and macrophage inflammatory protein-3 (MIP3a). In addition, corneas of PSP group had a more regular and compact architecture of collagen with thinner corneal thickness than in the AME group. ' CONCLUSION: Polysaccharide extract from Spirulina platensis inhibited alkali burn-induced inflammation and CorNV more effectively than AME Extract at the studied doses, thus may be used for the therapy of corneal diseases involving neovascularization and inflammation.
Ling-Ling YangQing-Jun ZhouYao WangYan Gaoand Yi-Qiang Wang
关键词:INFLAMMATION
羊膜对人视网膜色素上皮细胞增生和分化的影响
2012年
背景人视网膜色素上皮(RPE)细胞移植治疗视网膜变性性疾病是目前的研究热点之一,许多生物载体可制备RPE细胞单层,但多存在着细胞毒性、稳定性差及免疫反应等问题。目的检测羊膜对人RPE细胞增生和分化的影响,探讨其作为人RPE细胞层载体进行移植的可能性。方法将RPE细胞系ARPE-19细胞株用含质量分数10%胎牛血清的DMEM/F12培养基进行培养和传代,8~12代细胞用于实验。将传代细胞分为2个组,一组细胞接种于去上皮羊膜作为实验组,另一组细胞直接在培养孔内进行培养作为对照组。分别于接种后24、48、72、96h行MTT实验,测定RPE细胞的吸光度(A492)值以评估两组细胞在增生能力方面的不同;培养3周后的细胞层行苏木精-伊红染色,检测细胞在不同培养载体上是否存在形态学方面的区别;分别收集同一孔中生长于羊膜和培养板表面的细胞,采用逆转录聚合酶链反应(RT—PCR)法检测色素上皮衍生因子(PEDF)、钙黏蛋白(N—cadherin)、β-连环蛋白(β—catenin)以及细胞连接相关蛋白的表达情况;同时收集培养3周时的细胞行透射电子显微镜和扫描电子显微镜检查,比较不同载体培养的ARPE-19细胞在超微结构方面的区别。结果与对照组相比,实验组细胞增生的速度明显变缓,两组细胞增生情况比较差异有统计学意义(F=41.760,P=0.000)。苏木精-伊红染色结果显示,同一培养孔内实验组的细胞密度明显低于对照组,说明羊膜对ARPE-19细胞增生的影响不是通过细胞分泌的可溶性因子的作用。RT—PCR结果显示,实验组细胞连接相关蛋白claudin 1、N—cadherin和PEDF mRNA在培养细胞上的表达水平均明显高于对照组,差异均有统计学意义(t=15.828,P=0.000;t=6.839,P=0.002;t=14.667,P=0.000);Connexin 43的表达低于对照组,差异有统计学意义(t=3�
王瑶宫华青杨玲玲王茜周庆军王宜强
关键词:羊膜视网膜色素上皮细胞增生分化
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