Adtani Pooja Narain, Narasimhan Malathi, Punnoose Alan M, Kambalachenu Himavanth Reddy
Department of Oral Pathology and Microbiology, Faculty of Dental Sciences, Sri Ramachandra University and Research Institute, Chennai, India.
Center for Regenerative Medicine and Stem Cell Research, Central Research Facility, Sri Ramachandra University and Research Institute, Chennai, India.
J Investig Clin Dent. 2017 May;8(2). doi: 10.1111/jicd.12208. Epub 2016 Feb 3.
The aim of the present study was to investigate the in vitro antifibrogenic effects of Centella asiatica Linn (CA) and its bioactive triterpene aglycone asiatic acid (AA) on arecoline-induced fibrosis in primary human buccal fibroblasts (HBF).
An ethanolic extract of CA was prepared, and AA was purchased commercially. High-performance thin-layer chromatography (HPTLC) was performed to quantify AA in the CA extract; colorimetric assay (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) was performed to determine an half-maximal inhibitory concentration. HBF were cultured and stimulated with arecoline. The inhibitory effects of CA and AA at different concentrations were assessed using gene-expression studies on fibrosis-related markers: transforming growth factor-β1, collagen 1 type 2, and collagen 3 type 1. The stimulatory effect of arecoline and the inhibitory effect of AA on fibroblast morphology and extracellular matrix were assessed qualitatively using Masson trichrome stain.
The HPTLC analysis determined 1.2% AA per 100 g of CA extract. Arecoline produced a concentration-dependent increase in the fibrotic markers, treatment with CA significantly downregulated fibrotic markers at higher concentrations, and AA downregulated at lower concentrations. Arecoline altered fibroblast morphology and stained strongly positive for collagen, and AA treatment regained fibroblast morphology with faint collagen staining.
CA and AA can be used as antifibrotic agents.
本研究旨在探讨积雪草及其生物活性三萜苷元积雪草苷对槟榔碱诱导的原代人颊成纤维细胞（HBF）纤维化的体外抗纤维化作用。
制备积雪草乙醇提取物，积雪草苷购自商业渠道。采用高效薄层色谱法（HPTLC）对积雪草提取物中的积雪草苷进行定量；采用比色法（3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐）测定半数抑制浓度。培养HBF并用槟榔碱刺激。使用与纤维化相关标志物的基因表达研究评估不同浓度的积雪草和积雪草苷的抑制作用：转化生长因子-β1、I型胶原和III型胶原。使用Masson三色染色法定性评估槟榔碱的刺激作用以及积雪草苷对成纤维细胞形态和细胞外基质的抑制作用。
HPTLC分析确定每100 g积雪草提取物中积雪草苷含量为1.2%。槟榔碱使纤维化标志物呈浓度依赖性增加，高浓度的积雪草处理显著下调纤维化标志物，低浓度的积雪草苷下调纤维化标志物。槟榔碱改变了成纤维细胞形态，胶原染色呈强阳性，而积雪草苷处理使成纤维细胞形态恢复，胶原染色变淡。
积雪草和积雪草苷可作为抗纤维化药物。
