Fdu 540

Succulent and spatulate stems of OFI were provided by Biodiversity Research Institute, Jeju Technopark (voucher specimens No. 20080711006). and They were collected on Jeju Island, Republic of Korea (Latitude 33.24588, Longitude 126.23090), and were authenticated by Professor Geung-Joo Lee at Chungnam National University, Daejeon, Korea. The harvested stems were washed and freeze-dried for 48h in a freeze dryer (FD5518, Ilshin Co., Yangju, Korea). The dried OFI stems were ground using a grinder (SMX-4000DY, Shinil Co., Cheonan, Korea) for 1 min to pass through a100-mesh screen. The ground OFI stems were subjected to extraction with eight volumes of water, 30% ethanol, or 50% ethanol for 5 h at 60 °C. The extracts were then concentrated under reduced pressure using a rotary evaporator (Buchi R-114 Rotary evaporator with Buchi B-480 heating Bath, Flawil, Switzerland), suspended in distilled water to >20 Brix, and dried to obtain a powder by a freeze drier (Freeze dryer, FDU-540, Eyela, Tokyo, Japan).

The CP samples were purchased from Onggi Korean Herbal Medicine Market (Daegu, Korea) figurein April 2019, and herbal identification was performed by Professor Young-Cheol Lee, College of Korean Medicine, Sangji University, Wonju, Korea. A voucher specimen (no. 2019-SJCP-1) was deposited in the laboratory of the Department of Herbology, College of Korean Medicine, Sangji University Wonju 26339, Republic of Korea. Dried and chopped CP (600 g) was extracted three times with 70% ethanol using a 3 h reflux. The materials were filtered under reduced pressure at 40 °C using a vacuum rotatory evaporator (BUCHI B-480, Buchi, Flawil, Switzerland) and dried in a freeze-drier (EYELA FDU-540, Japan) to yield the CP extract (29.56 g). The yield (w/w) of the extract was approximately 4.93%. Dexamethasone (Sigma Aldrich, Korea), which was dissolved in saline, was used as a positive control. Methacholine, OVA, OA, and aluminum hydroxide were purchased from Sigma-Aldrich. All other chemicals and solvents used in the experiments were of analytical grade and were purchased from Sigma-Aldrich, unless otherwise indicated.

The samples of HD were purchased from Omniherb Korean Herbs Co. Ltd. (Daegu, Korea) in May 2016, and identified by Professor Young-Cheol Lee, College of Korean Medicine, Sangji University in Wonju, Korea. Voucher specimens (no. 2016-SJHD-1) were deposited in our laboratory at the Department of Herbology, College of Korean Medicine, Sangji University Wonju 220-702, Republic of Korea. Dried and chopped HD (1 kg) was refluxed three times with 70% ethanol for 8 h. The materials were filtered under reduced pressure at 40 °C by using a vacuum rotatory evaporator (BUCHI B-480, Flawil, Switzerland) and dried in a freeze-drier (EYELA FDU-540, Tokyo, Japan) to yield the HD extract (114.57 g). The yield (w/w) of the extract was approximately 11.46%. OVA, methacholine, and aluminum hydroxide were purchased from Sigma. All other chemical reagents were analytical grade and were purchased from Sigma, unless otherwise indicated.