Liu Zibin, Zhang Rui, Lai Jian
Department of Ophthalmology, Hangzhou TCM Hospital Affiliated to Zhejiang Chinese Medical University, No. 453, Stadium Road, Hangzhou, 310007 Zhejiang China.
Cytotechnology. 2025 Jun;77(3):105. doi: 10.1007/s10616-025-00769-9. Epub 2025 May 20.
Meibomian gland dysfunction (MGD) is an ocular surface disease lacking optimal treatment strategy. The Hedgehog pathway is involved in regulating MGEC proliferation and differentiation. Catalpol (CAT) is the main active ingredient in with therapeutic potential. Exploring the effects and biological mechanisms of CAT on meibomian gland epithelial cells (MGECs). Primarily cultured rat MGECs were co-cultured with 3T3 cells for 7 days. MGECs were exposed to 2.5, 5, and 10 mmol/L CAT, 10 μg/mL Azithromycin (AZM), and 0.6 μmol/L Smoothened receptor agonist (SAG) for 48 h. Colony formation assays, Cell counting kit-8, Ki67 immunofluorescence, Nile red and Oil red O staining, and HSD LipidTOX Green kits were used to assess cell proliferation and lipid accumulation. Real-time quantitative PCR and Western blot analysis were used to measure gene expressions related to Hedgehog- and peroxisome proliferator-activated receptor (PPAR)-γ. This study successfully isolated primarily rat MGECs (expressed P63 and K14). AZM and 5, and 10 mmol/L CAT inhibited colony number, cell viability, and Ki67 mean fluorescence intensity (MFI), while they enhanced MFI of Nile red and LipidTOX Green, as well as increasing the ratio of Oil red O staining area. Additionally, transcription and translation levels of the Hedgehog pathway were significantly suppressed, meanwhile, PPAR-γ and SREBP-1 expression were increased. Interestingly, SAG reversed the effects of 10 mmol/L CAT on MGECs. CAT suppresses MGEC proliferation and promotes lipid accumulation by inhibiting the Hedgehog signaling pathway. This study offers a potential therapeutic strategy for MGD.
The online version contains supplementary material available at 10.1007/s10616-025-00769-9.
睑板腺功能障碍（MGD）是一种缺乏最佳治疗策略的眼表疾病。刺猬信号通路参与调节睑板腺上皮细胞（MGEC）的增殖和分化。梓醇（CAT）是具有治疗潜力的主要活性成分。探索CAT对睑板腺上皮细胞（MGECs）的作用及生物学机制。将原代培养的大鼠MGECs与3T3细胞共培养7天。将MGECs分别暴露于2.5、5和10 mmol/L的CAT、10 μg/mL阿奇霉素（AZM）和0.6 μmol/L的 smoothened 受体激动剂（SAG）中48小时。采用集落形成试验、细胞计数试剂盒-8、Ki67免疫荧光、尼罗红和油红O染色以及HSD LipidTOX Green试剂盒评估细胞增殖和脂质积累。采用实时定量PCR和蛋白质印迹分析来检测与刺猬信号通路和过氧化物酶体增殖物激活受体（PPAR）-γ相关的基因表达。本研究成功分离出原代大鼠MGECs（表达P63和K14）。AZM以及5和10 mmol/L的CAT抑制了集落数量、细胞活力和Ki67平均荧光强度（MFI），同时增强了尼罗红和LipidTOX Green的MFI，并增加了油红O染色面积的比例。此外，刺猬信号通路的转录和翻译水平被显著抑制，同时PPAR-γ和SREBP-1的表达增加。有趣的是，SAG逆转了10 mmol/L CAT对MGECs的影响。CAT通过抑制刺猬信号通路抑制MGEC增殖并促进脂质积累。本研究为MGD提供了一种潜在的治疗策略。
在线版本包含可在10.1007/s10616-025-00769-9获取的补充材料。
