Anti-Inflammatory and Anti-Apoptotic Effects of Acer Palmatum Thumb. Extract, KIOM-2015EW, in a Hyperosmolar-Stress-Induced In Vitro Dry Eye Model
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials and Reagents
2.2. Preparation of Herbal Extract, KIOM-2015EW
2.3. HCEC Culture
2.4. Cell-Viability Assay
2.5. ELISA
2.6. RNA Isolation and Reverse Transcription–Polymerase Chain Reaction (RT-PCR) Analysis
2.7. Cellular Protein Extraction and Western Blotting
2.8. Immunofluorescence Staining
2.9. Flow Cytometry
2.10. Cytoplasmic and Nuclear Protein Extraction
2.11. High Performance Liquid Chromatography (HPLC) Analysis
2.12. Statistical Analysis
3. Results
3.1. Effects of Hyperosmotic Medium and KIOM-2015EW on HCECs
3.2. KIOM-2015EW Significantly Decreased Pro-Inflammatory Cytokine Expression in HOS-Exposed HCECs
3.3. KIOM-2015EW Blocked Apoptosis Due to HOS-Induced Cytotoxicity in HCECs
3.4. KIOM-2015EW Downregulates HOS-Mediated p38, ERK, and JNK Activation in HCECs
3.5. MAPK Inhibition by KIOM-2015EW Blocks HOS-Induced Apoptosis
3.6. KIOM-2015EW Suppresses HOS-Induced NF-κB Activation through p65 Nuclear Translocation Blockage in HCECs
3.7. Identification of the Main Components in KIOM-2015EW Using HPLC
3.8. Orientin, Isoorientin and Vitexin Reduced Level of HOS-Induced Pro-Inflammatory Cytokines
4. Discussion
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Primers | ||||
---|---|---|---|---|
Gene Name | Sequence | Tm. (°C) | Cycles | |
TNF-α | F | 5′-GCGGTGCTTGTTCCTCAG-3′ | 58 | 40 |
R | 5′-CTGGCAGGGGCTCTTGAT-3′ | |||
IL-1β | F | 5′-TCAGCACCTCTCAAGCAGAA-3′ | 58 | 30 |
R | 5′-ATCAGAATGTGGGAGCGAATG-3′ | |||
IL-6 | F | 5′-CAATCTGGATTCAATGAGGAGACTT-3′ | 58 | 30 |
R | 5′-CCATTAACAACAACAATCTGAGGTG-3′ | |||
GAPDH | F | 5′-GAAGGTGAAGGTCGGAGT-3′ | 56 | 35 |
R | 5′-GAAGATGGTGATGGGATTTC-3′ |
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Kim, Y.-H.; Oh, T.W.; Park, E.; Yim, N.-H.; Park, K.I.; Cho, W.K.; Ma, J.Y. Anti-Inflammatory and Anti-Apoptotic Effects of Acer Palmatum Thumb. Extract, KIOM-2015EW, in a Hyperosmolar-Stress-Induced In Vitro Dry Eye Model. Nutrients 2018, 10, 282. https://doi.org/10.3390/nu10030282
Kim Y-H, Oh TW, Park E, Yim N-H, Park KI, Cho WK, Ma JY. Anti-Inflammatory and Anti-Apoptotic Effects of Acer Palmatum Thumb. Extract, KIOM-2015EW, in a Hyperosmolar-Stress-Induced In Vitro Dry Eye Model. Nutrients. 2018; 10(3):282. https://doi.org/10.3390/nu10030282
Chicago/Turabian StyleKim, Yeoun-Hee, Tae Woo Oh, Eunhee Park, Nam-Hui Yim, Kwang Il Park, Won Kyung Cho, and Jin Yeul Ma. 2018. "Anti-Inflammatory and Anti-Apoptotic Effects of Acer Palmatum Thumb. Extract, KIOM-2015EW, in a Hyperosmolar-Stress-Induced In Vitro Dry Eye Model" Nutrients 10, no. 3: 282. https://doi.org/10.3390/nu10030282