Yusof Alia Md, Abd Ghafar Norzana, Kamarudin Taty Anna, Hui Chua Kien, Yusof Yasmin Anum Mohd
Department of Anatomy, Universiti Kebangsaan Malaysia Medical Centre (UKMMC), Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000, Kuala Lumpur, Malaysia.
Department of Physiology, Universiti Kebangsaan Malaysia Medical Centre (UKMMC), Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000, Kuala Lumpur, Malaysia.
BMC Complement Altern Med. 2016 Feb 24;16:76. doi: 10.1186/s12906-016-1055-7.
This study aimed to evaluate the effects of Gelam honey on corneal keratocytes proliferative capacity and phenotypic characterization via MTT assay, gene expression and immunocytochemistry.
Corneal keratocytes from New Zealand white rabbits were cultured in basal medium (BM) and serum enriched medium (BMS). Serial dilutions of Gelam honey (GH) were added to both media and cells were cultured until passage 1. MTT assay was performed on corneal keratocytes in both media to ascertain the optimal dose of GH that produced maximum proliferation.
Gelam honey at the concentration of 0.0015% in both media showed the highest proliferative capacity with no morphological changes compared to their respective controls. The gene expression of aldehyde dehydrogenase (ALDH), a marker for quiescent keratocytes and vimentin, a marker for fibroblast, were higher in the GH enriched groups. The alpha smooth muscle actin (α-SMA) expression, marker for myofibroblast, was lower in GH treated groups compared to the controls. Immunocytochemistry results were in accordance to the gene expression analyses.
Gelam honey at a concentration of 0.0015% promotes ex vivo corneal keratocytes proliferation while retaining desirable phenotype expression. The results serve as a basis for the development of Gelam honey as a potential natural product in promoting corneal wound healing.
本研究旨在通过MTT法、基因表达和免疫细胞化学评估吉拉蜂蜜对角膜基质细胞增殖能力和表型特征的影响。
将新西兰白兔的角膜基质细胞培养于基础培养基(BM)和富血清培养基(BMS)中。将吉拉蜂蜜(GH)的系列稀释液添加到两种培养基中,细胞培养至第1代。对两种培养基中的角膜基质细胞进行MTT法检测,以确定产生最大增殖的GH最佳剂量。
与各自的对照组相比,两种培养基中浓度为0.0015%的吉拉蜂蜜显示出最高的增殖能力,且无形态学变化。静息角膜基质细胞标志物醛脱氢酶(ALDH)和成纤维细胞标志物波形蛋白的基因表达在富含GH的组中较高。与对照组相比,GH处理组中肌成纤维细胞标志物α平滑肌肌动蛋白(α-SMA)的表达较低。免疫细胞化学结果与基因表达分析一致。
浓度为0.0015%的吉拉蜂蜜可促进离体角膜基质细胞增殖,同时保留理想的表型表达。这些结果为将吉拉蜂蜜开发为促进角膜伤口愈合的潜在天然产物奠定了基础。