Comprehensive Evaluation of the Anti-Helicobacter pylori Activity of Scutellariae Radix

Ba Wool Lee; II-Ho Park; Dongsool Yim; Sung Sook Choi

doi:10.20307/nps.2017.23.1.46

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Lee, Park, Yim, and Choi: Comprehensive Evaluation of the Anti-Helicobacter pylori Activity of Scutellariae Radix

Original Article

Natural Product Sciences 2017;23(1):46-52.

Published online: 20 January 2017

DOI: https://doi.org/10.20307/nps.2017.23.1.46

Comprehensive Evaluation of the Anti-Helicobacter pylori Activity of Scutellariae Radix

Ba Wool Lee¹, II-Ho Park¹, Dongsool Yim^1,², Sung Sook Choi^1,^∗

¹College of Pharmacy, Sahmyook University, 815, Hwarang-ro, Nowon-gu, Seoul, 01795 Republic of Korea

²Re-creation Institute of Phytochemicals, College of Pharmacy, Sahmyook University, 815, Hwarang-ro, Nowon-gu, Seoul 01795, Republic of Korea

∗Author for correspondence SungSook Choi, College of Pharmacy, Sahmyook University, 815, Hwarang-ro, Nowon-gu, Seoul 01795, Republic of Korea. Tel: +82-2-3399-1606; E-mail: sschoi@syu.ac.kr

Received 22 September 2016 Revised 10 January 2017 Accepted 12 January 2017

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The aim of this study was to evaluate the anti-Helicobacter pylori activity of fractions and major aglycon compounds (baicalein, chrysin, oroxylin A, wogonin) of Scutellariae Radix. Minimum inhibitory concentration (MIC) measurement, DPPH radical-scavenging assay, DNA protection assay, and urease inhibition analysis were performed. The ethyl acetate (EtOAc) fraction showed the potent anti-Helicobacter activity, and therefore, compounds in the EtOAc fraction were subjected to further assay. The MICs of chrysin, oroxylin A, and wogonin against Helicobacter pylori 26695 were 6.25, 12.5 and 25 µg/mL, respectively. Baicalein exhibited the most effective DPPH radical-scavenging activity. DNA protection using Fenton reaction, chrysin, oroxylin A, and wogonin showed effective DNA protective effect. This result was also confirmed by quantitative real-time polymerase chain reaction (qRT-PCR). Regarding Jack bean urease (0.5 mg/mL, 50 unit/mg) inhibition, 20 mM ofbaicalein and chrysin inhibited urease activity by 88.2% and 72.5%, respectively.

Keywords: Scutellariae Radix, Helicobacter pylori, MIC, DNA protection assay, Urease inhibitory activity

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Fig. 1.

Chemical Structures of flavonoids from Scutellariae Radix.

Fig. 2.

Chromatogram of five standards (baicalin, baicalein, wogonin, oroxylin A, chrysin) and each fraction of Scutellariae Radixby HPLC analysis. A: five standards; B: total extract; C: EtOAc fraction; D: n-butanol fraction.

Fig. 3.

DPPH radicalscavenging activity of fractionsand compounds from ScutellariaeRadix. Each fraction (A) (0.25 – 2.0 mg/ mL) and compound (B) (0.25 – 1.0 mM) was tested and ascorbic acid and BHA (0.25 mg/mL for fractions, 1 mM for compounds) were used as positive controls. Data represent the mean ± SD of three replicates; ∗p < 0.05, ∗∗p < 0.01, significant difference compared to the negative control.

Fig. 4.

DNA protective effect against hydrogen peroxide of fractions (A) and compounds (B) from Scutellaria Radix. (A) 1: AGS cell; 2: AGS cell + Fenton reagent; 3: AGS cell + Fenton reagent + quercetin; 4: AGS cell + Fenton reagent + total Fr.; 5: AGS cell + Fenton reagent + EtOAc Fr.; 6: AGS cell + Fenton reagent + n-BuOH Fr. (B)1: AGS cell; 2: AGS cell + Fenton reagent; 3: AGS cell + Fenton reagent + quercetin; 4: AGS cell + Fenton reagent+bacialein; 5: AGS cell+Fenton reagent+chrysin; 6: AGS cell + Fenton reagent + oroxylin A; 6: AGS cell + Fenton reagent + wogonin. The concentrations of each fraction and each compound were 0.25 mg/mL and 1 mM respectively.

Fig. 5.

H. pylori urease inhibitory activity of fractions (A) and compounds (B) from Scutellariae Radix. Data represent the means ± SD of three replicates; ∗p < 0.05, ∗∗p < 0.01, significant difference compared to the negative control.

Fig. 6.

Jack bean urease inhibitory activity of four compounds from Scutellariae Radix. Data represent the means ± SD of three replicates; ∗p < 0.05, ∗∗p < 0.01, significant difference compared to the negative control.

Table 1.

Anti-Helicobacter activity of fractions and compounds from Scutellria Radix

Samples		H. pylori 26695
Samples		MIC^a (µg/mL)	MBC^b (µg/mL)
	Total	125	250
Fractions	EtOAc	31.25	31.25
	BuOH	125	125
	Baicalin	>250	>250
	Baicalein	50	50
Compounds	Chrysin	6.25	12.5
	Oroxylin A	12.5	25
	Wogonin	25	25

^a Minimum inhibitory concentration

^b Minimum bactericidal concentration

Table 2.

Quantitative analysis of DNA protective effect of fractions and compounds from Scutellria Radix

Groups	1	2	3	4	5	6	7
Fractions	1^a	0.523 ± 0.026	0.841 ± 0.047	0.751 ± 0.036	0.834 ± 0.067	0.864 ± 0.038	−
Compounds	1	0.490 ± 0.025	0.736 ± 0.036	0.637 ± 0.019	0.746 ± 0.046	0.777 ± 0.054	0.775 ± 0.038

^a relative band intensity of each sample compared with control (AGS cell). 1: Control (AGS cell); 2: AGS cell + Fenton reagent; 3: AGS cell + Fenton reagent + quercetin; 4: AGS cell + Fenton reagent + total Fr./ bacalein; 5: AGS cell + Fenton reagent + EtOAc Fr./ chrysin; 6: AGS cell + Fenton reagent + n-BuOH Fr./ oroxylin A; 7: AGS cell + Fenton reagent + wogonin.

Table 3.

qRT-PCR analysis of AGS cell DNA damaged by Fenton reaction.

DNA Samples	Compounds	Ct value^a
AGS cell	none	11.437 ± 0.234
AGS cell + Fenton reagents	none	29.799 ± 0.215
AGS cell + Fenton reagents	Quercetin	12. 352 ± 0.324
AGS cell + Fenton reagents	Baicalein	20.799 ± 0.215
AGS cell + Fenton reagents	Chrysin	16. 877 ± 0.674
AGS cell + Fenton reagents	Oroxylin A	17.719 ± 0.189
AGS cell + Fenton reagents	Wogonin	14.417 ± 0.089

^a ct value or threshold cycle value is the cycle number at which the fluorescence generated within a reaction crosses the fluorescence threshold. Ct value is inversely proportional to the detectable amount of amplicon product.

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