Use of human Caco-2 cells and HPAE-PAD for α-glucosidase assay

Yuki Ikeda; So Nishimoto; Ying Qiao; Haruna Yano; Hideaki Minami; Masaaki Ito; Toshiyuki Kimura; Teisuke Takita; Kiyoshi Yasukawa

doi:10.1016/j.vascn.2024.107508

Use of human Caco-2 cells and HPAE-PAD for α-glucosidase assay

J Pharmacol Toxicol Methods. 2024 Apr 24:127:107508. doi: 10.1016/j.vascn.2024.107508. Online ahead of print.

Authors

Yuki Ikeda¹, So Nishimoto¹, Ying Qiao¹, Haruna Yano¹, Hideaki Minami², Masaaki Ito³, Toshiyuki Kimura⁴, Teisuke Takita¹, Kiyoshi Yasukawa⁵

Affiliations

¹ Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.
² Kyoto Municipal Institute of Industrial Technology and Culture, Shimogyo-ku, Kyoto 600-8815, Japan.
³ Department of Bioresources Engineering, National Institute of Technology, Okinawa College, Henoko, Nago 905-2192, Japan.
⁴ Research Center of Advanced Analysis, National Agriculture and Food Research Organization (NARO), Tsukuba 305-8642, Japan.
⁵ Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan. Electronic address: yasukawa.kiyoshi.7v@kyoto-u.ac.jp.

PMID: 38670388
DOI: 10.1016/j.vascn.2024.107508

Abstract

To measure α-glucosidase activity, rat intestinal acetone powder is commonly used as a source of α-glucosidase, and the mutarotase-glucose oxidase (GOD) methods commonly used to quantitate glucose produced by enzymatic hydrolysis of the substrates. In this study, we compared human Caco-2 cell extracts with rat intestinal acetone powder extracts. We also compared high-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD) with the mutarotase-GOD method. The sensitivity of HPAE-PAD was higher than that of mutarotase-GOD. The glucose concentration quantified by HPAE-PAD was similar to that quantified using the mutarotase-GOD method. In the maltase reaction, 1-deoxynojirimycin (1-DNJ) exerted a more potent inhibitory effect on human enzymes than on rat enzymes. This order was reversed during the sucrase reaction. These results suggested that the combined use of Caco-2 cell extracts and HPAE-PAD is advantageous for use in α-glucosidase-related basic research.

Keywords: Caco-2 cell; HPAE-PAD; Mutarotase-GOD method; Rat intestinal acetone powder; α-Glucosidase.