Yan-Xu Chang1, Ai-Hua Ge2, Sineeporn Donnapee3, Jin Li3, Yang Bai4, Jiao Liu4, Jun He3, Xi Yang3, Li-Jiao Song4, Bo-Li Zhang3, Xiu-Mei Gao5. 1. Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; Tianjin Key Laboratory of Phytochemistry and Pharmaceutical Analysis, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China. Electronic address: Tcmcyx@126.com. 2. Tianjin Key Laboratory of Phytochemistry and Pharmaceutical Analysis, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China. 3. Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; Tianjin Key Laboratory of Phytochemistry and Pharmaceutical Analysis, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China. 4. Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China. 5. Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; Tianjin Key Laboratory of Phytochemistry and Pharmaceutical Analysis, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China. Electronic address: gaoxiumei@tjutcm.edu.cn.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: Jinqi Jiangtang Tablet is a traditional Chinese anti-diabetic formula containing three ingredients: Coptis chinensis Franch. (dried rhizome of C. chinensis Franch., Coptis deltoidea C. Y. Cheng et Hsiao and Coptis teeta Wall.), Astragalus membranaceus (Fisch.) Bunge. (dried root of A. membranaceus (Fisch.) Bge. var. mongholicus (Bge. ) Hsiao and A. membranaceus (Fisch.) Bge. ) and Lonicera japonica Thunb. (dried alabastrum or with nascent flowers of L. japonica Thunb. ). Free radicals, α-glucosidase, α-amylase, aldose reductase and lipase are different targets related with diabetes. However, there are no chromatographic methods employed in screening the anti-diabetic compounds from natural products basing on these targets simultaneously. The present study was aimed at the establishment of a multi-targets integrated fingerprinting to clarify the possible mechanism of the action of Traditional Chinese Medicines which simultaneously contained multiple chemical characteristics and effects of constitutions. MATERIALS AND METHODS: The multi-targets integrated fingerprinting was developed and validated to screen anti-diabetic compounds from natural products by using ultra-high-performance liquid chromatography/quadruple-time-of-flight mass spectrometry, fraction collector and microplate reader. Ultra performance liquid chromatography was employed to separate the components in Jinqi Jiangtang Tablet, which were identified by quadruple-time-of-flight mass spectrometry to acquire their structural information and collected by the fraction collector. Finally the active fractions were tested for scavenging 1, 1-diphenyl-2-picrylhydrazyl radical and inhibition of α-glucosidase, α-amylase, aldose reductase, and lipase activities in vitro by microplate reader. RESULTS: Our tests revealed that the Jinqi Jiangtang Tablet showed inhibitory activity against α-glucosidase, α-amylase, aldose reductase and lipase with IC50 values of 0.80 ± 0.02 mg/mL, 1.28 ± 0.13 mg/mL, 0.80 ± 0.02 mg/mL, 1.90 ± 0.18 mg/mL respectively and the scavenging activity with IC50 value of 1.71 ± 0.178 mg/mL. The bioactive fractions were identified to be alkaloids, flavonoids and phenolic acids. The phenolic acids possessed antioxidant activities, namely the scavenging effect on 1, 1-diphenyl-2-picrylhydrazyl rull;). The alkaloids exhibited inhibitory activity against α-glucosidase, aldose reductase, α-amylase, and lipase. The flavonoids also showed mild inhibition on α-glucosidase, aldose reductase, α-amylase and lipase. CONCLUSIONS: The results demonstrate that Jinqi Jiangtang Tablet can scavenge free radicals and inhibit α-glucosidase, aldose reductase, α-amylase and lipase, which may be the possible mechanism of action of Jinqi Jiangtang Tablet for the treatment of diabetes and associated complications. Compared with conventional chromatographic separation and activity assays, the multi-targets integrated fingerprinting, which simultaneously contains the chemical characteristics and multiple effects of constitutions could comprehensively and properly reveal the activity of Jinqi Jiangtang Tablet. The results also show that the multi-targets integrated fingerprinting is a novel and powerful tool for screening and identifying active ingredients from Traditional Chinese Medicines.
ETHNOPHARMACOLOGICAL RELEVANCE: Jinqi Jiangtang Tablet is a traditional Chinese anti-diabetic formula containing three ingredients: Coptis chinensis Franch. (dried rhizome of C. chinensis Franch., Coptis deltoidea C. Y. Cheng et Hsiao and Coptis teeta Wall.), Astragalus membranaceus (Fisch.) Bunge. (dried root of A. membranaceus (Fisch.) Bge. var. mongholicus (Bge. ) Hsiao and A. membranaceus (Fisch.) Bge. ) and Lonicera japonica Thunb. (dried alabastrum or with nascent flowers of L. japonica Thunb. ). Free radicals, α-glucosidase, α-amylase, aldose reductase and lipase are different targets related with diabetes. However, there are no chromatographic methods employed in screening the anti-diabetic compounds from natural products basing on these targets simultaneously. The present study was aimed at the establishment of a multi-targets integrated fingerprinting to clarify the possible mechanism of the action of Traditional Chinese Medicines which simultaneously contained multiple chemical characteristics and effects of constitutions. MATERIALS AND METHODS: The multi-targets integrated fingerprinting was developed and validated to screen anti-diabetic compounds from natural products by using ultra-high-performance liquid chromatography/quadruple-time-of-flight mass spectrometry, fraction collector and microplate reader. Ultra performance liquid chromatography was employed to separate the components in Jinqi Jiangtang Tablet, which were identified by quadruple-time-of-flight mass spectrometry to acquire their structural information and collected by the fraction collector. Finally the active fractions were tested for scavenging 1, 1-diphenyl-2-picrylhydrazyl radical and inhibition of α-glucosidase, α-amylase, aldose reductase, and lipase activities in vitro by microplate reader. RESULTS: Our tests revealed that the Jinqi Jiangtang Tablet showed inhibitory activity against α-glucosidase, α-amylase, aldose reductase and lipase with IC50 values of 0.80 ± 0.02 mg/mL, 1.28 ± 0.13 mg/mL, 0.80 ± 0.02 mg/mL, 1.90 ± 0.18 mg/mL respectively and the scavenging activity with IC50 value of 1.71 ± 0.178 mg/mL. The bioactive fractions were identified to be alkaloids, flavonoids and phenolic acids. The phenolic acids possessed antioxidant activities, namely the scavenging effect on 1, 1-diphenyl-2-picrylhydrazyl rull;). The alkaloids exhibited inhibitory activity against α-glucosidase, aldose reductase, α-amylase, and lipase. The flavonoids also showed mild inhibition on α-glucosidase, aldose reductase, α-amylase and lipase. CONCLUSIONS: The results demonstrate that Jinqi Jiangtang Tablet can scavenge free radicals and inhibit α-glucosidase, aldose reductase, α-amylase and lipase, which may be the possible mechanism of action of Jinqi Jiangtang Tablet for the treatment of diabetes and associated complications. Compared with conventional chromatographic separation and activity assays, the multi-targets integrated fingerprinting, which simultaneously contains the chemical characteristics and multiple effects of constitutions could comprehensively and properly reveal the activity of Jinqi Jiangtang Tablet. The results also show that the multi-targets integrated fingerprinting is a novel and powerful tool for screening and identifying active ingredients from Traditional Chinese Medicines.
Authors: Weiyan Yin; Tengfei Li; Sheng-Che Hung; Han Zhang; Li Wang; Dinggang Shen; Hongtu Zhu; Peter J Mucha; Jessica R Cohen; Weili Lin Journal: Proc Natl Acad Sci U S A Date: 2020-08-31 Impact factor: 11.205