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白绵毛柳穿鱼提取物对培养的人角膜上皮细胞(10.014 pRSV-T)的影响

The Effect of Lamium album Extract on Cultivated Human Corneal Epithelial Cells (10.014 pRSV-T).

作者信息

Paduch Roman, Woźniak Anna

机构信息

Department of Virology and Immunology, Institute of Microbiology and Biotechnology, Maria Curie-Skłodowska University, Lublin, Poland.

Department of General Ophthalmology, Medical University of Lublin, Lublin, Poland.

出版信息

J Ophthalmic Vis Res. 2015 Jul-Sep;10(3):229-37. doi: 10.4103/2008-322X.170349.

Abstract

PURPOSE

To evaluate the effect of Lamium album extract on human corneal epithelial cells (10.014 pRSV-T cell line) cultured in vitro.

METHODS

Normal human corneal epithelial cells were incubated with ethanol, ethyl acetate and heptane extracts from Lamium album. Their effect on cells was evaluated by neutral red (NR) uptake and MTT assays for cytotoxicity, ELISA for immunomodulation, Griess method for nitric oxide levels, DPPH assay for free radicals scavenging activity. A blank control consisted only of culture medium.

RESULTS

In NR and MTT assays, Lamium album extracts did not affect cell viability (80% at 125 μg/ml concentration). Ethanol was the least toxic extract (cell viability over 88%) and expressed the most potent reactive oxygen species (ROS) scavenging action. It was 19.88 ± 0.87% higher than controls representing a reduction corresponding to 7.136 μg/ml of trolox. Heptane extract revealed no ROS scavenging activity. All extracts decreased NO production by cells. The most active extract was ethanol (8 μg/ml) which reduced NO level to 0.242 μM (75% decrease compared to control). Extracts influenced pro-inflammatory (IL-1, IL-6, TNF-α) and anti-inflammatory (IL-10) cytokines levels reducing all of them in general. The strongest reduction in tested cytokines level was observed by the heptane extract. On the other hand, the ethanol extract induced mainly TNF-α level in a concentration dependent manner.

CONCLUSION

Selected Lamium album extracts influence human corneal epithelial cells. Generally, while not toxic, they modulate pro-inflammatory and anti-inflammatory cytokines levels, and decrease NO release by cells; moreover, ethanol and ethyl acetate extracts reduce ROS levels.

摘要

目的

评估白绵毛酸模提取物对体外培养的人角膜上皮细胞(10.014 pRSV - T细胞系)的作用。

方法

将正常人角膜上皮细胞与白绵毛酸模的乙醇提取物、乙酸乙酯提取物和庚烷提取物一起孵育。通过中性红(NR)摄取和MTT法检测细胞毒性来评估它们对细胞的作用,通过ELISA法检测免疫调节作用,通过Griess法检测一氧化氮水平,通过DPPH法检测自由基清除活性。空白对照仅由培养基组成。

结果

在NR和MTT检测中,白绵毛酸模提取物不影响细胞活力(浓度为125μg/ml时细胞活力达80%)。乙醇是毒性最小的提取物(细胞活力超过88%),并表现出最强的活性氧(ROS)清除作用。它比对照组高19.88±0.87%,相当于7.136μg/ml的Trolox的减少量。庚烷提取物未显示出ROS清除活性。所有提取物均降低了细胞产生的NO。最具活性的提取物是乙醇(8μg/ml),它将NO水平降低至0.242μM(与对照组相比降低了75%)。提取物影响促炎(IL - 1、IL - 6、TNF -α)和抗炎(IL - 10)细胞因子水平,总体上降低了所有这些细胞因子的水平。在测试的细胞因子水平上,观察到庚烷提取物的降低作用最强。另一方面,乙醇提取物主要以浓度依赖的方式诱导TNF -α水平。

结论

所选的白绵毛酸模提取物影响人角膜上皮细胞。一般来说,它们虽无毒,但可调节促炎和抗炎细胞因子水平,并减少细胞释放NO;此外,乙醇和乙酸乙酯提取物可降低ROS水平。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/54be/4687254/e189d0051070/JOVR-10-229-g001.jpg

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