Nat Prod Sci.  2017 Sep;23(3):175-182. 10.20307/nps.2017.23.3.175.

Anti-Helicobacter and Anti-inflammatory Effects of Sohamhyungtang in Helicobacter pylori-Infected Human Gastric Epithelial AGS cells

Affiliations
  • 1College of Pharmacy, Sahmyook University, 815, Hwarang-ro, Nowon-gu, Seoul 01795, Republic of Korea. sschoi@syu.ac.kr
  • 2Qu-best consulting, School of Dentistry, Seoul National University, Gwanak-ro, Gwanak-gu, Seoul 08826, Republic of Korea.
  • 3Re-creation Institute of Phytochemicals, College of Pharmacy, Sahmyook University, 815, Hwarang-ro, Nowon-gu, Seoul 01795, Republic of Korea.

Abstract

This study evaluated the anti-Helicobacter and anti-inflammatory effects of Sohamhyungtang (SHHT). The minimum inhibitory concentration (MIC) of SHHT against Helicobacter pylori (H. pylori) was determined by the agar dilution method. Expression of the H. pylori cagA gene in the presence of SHHT was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Inhibition of H. pylori urease by SHHT was determined by the phenol-hypochlorite assay. Antiadhesion activity of SHHT was measured by ureaphenol red reagent. Inhibition of nitric oxide (NO) production in AGS cells was measured with Griess reagent. Inducible nitric oxide synthase (iNOS) and IL-8 mRNA expression in AGS cells which were infected with H. pylori was determined by qRT-PCR. IL-8 level was measured by enzyme-linked immunosorbent assay (ELISA). The MIC of SHHT was 100 µg/mL and the expression of cagA gene was decreased about 25 folds in the presence of SHHT. H. pylori urease was inhibited 90% by SHHT. SHHT inhibited H. pylori adhesion on AGS cell in a concentration dependent manner. mRNA expression of iNOS and IL-8 and the production of NO and IL-8 were significantly decreased in the presence of SHHT. In conclusion, SHHT showed anti-Helicobacter activity and has potent anti-inflammatory effect on H. pylori-induced inflammation in human gastric epithelial AGS cells.

Keyword

Anti-helicobacter; Anti-inflammatory; Sohamhyungtang; CagA; AGS cell; IL-8

MeSH Terms

Agar
Enzyme-Linked Immunosorbent Assay
Helicobacter pylori
Helicobacter*
Humans*
Inflammation
Interleukin-8
Methods
Microbial Sensitivity Tests
Nitric Oxide
Nitric Oxide Synthase Type II
Real-Time Polymerase Chain Reaction
RNA, Messenger
Urease
Agar
Interleukin-8
Nitric Oxide
Nitric Oxide Synthase Type II
RNA, Messenger
Urease

Figure

  • Fig. 1 AGS cell viability test in the presence of various concentrations of SHHT (60~100 µg/mL), CRTA (10~50 µg/mL), and BRBR (7~12.5 µg/mL) for 24 hr. Changes in survival rate are presented as percentages of the control. Values are expressed as the mean ± SE of triplicate tests. *p < 0.05, **: p < 0.01, ***: p < 0.001. vs. control by ANOVA and Dunnett's multiple comparison test. Control: AGS cells, SHHT: AGS cells + Sohamhyungtang, CRTA: AGS cells + total alkaloid of rhizome of Coptidis Rhizoma, BRBR: AGS cells + Berberine.

  • Fig. 2 Inhibition of H. pylori urease in the presence of SHHT, CRTA, and BRBR. Changes in inhibition are presented as percentages of the control (100mM thiourea). Values are expressed as the mean ± SE of triplicate test. SHHT: Sohamhyungtang, CRTA: Total alkaloid of rhizome of Coptidis Rhizoma, BRBR: Berberine.

  • Fig. 3 Antiadhesion of SHHT, CRTA and BRBR against H. pylori to AGS human gastric cells. Antiadhesive activity was determined by urea phenol red method. The positive control means without samples and used to establish 100% attachment. The results are the mean ± SE for three independent experiments. *p < 0.05, **: p < 0.01, ***: p < 0.001. vs. control by ANOVA and Dunnett's multiple comparison test.

  • Fig. 4 Effects of SHHT, CRTA, and BRBR on H. pylori 26695-induced NO production in AGS cells. Changes in NO production are presented as percentages of control. Values are expressed as the mean ± S. E in triplicate tests. **: p < 0.01, ***: p < 0.001. vs. control by ANOVA and Dunnett's multiple comparison test. None: AGS cells, Control: AGS cells + H. pylori, SHHT: AGS cells + H. pylori + Sohamhyungtang, CRTA: AGS cells + H. pylori + Total alkaloid of rhizome of Coptidis Rhizoma, BRBR: AGS cells + H. pylori + Berberine. The effects of SHHT, CRTA, and BRBR itself on the AGS cells, they showed no difference compared with AGS cells (data was not shown).

  • Fig. 5 Effects of SHHT, CRTA, and BRBR on H. pylori 26695-induced IL-8 production in AGS cells. Changes in IL-8 production are presented as percentages of the control. Values are expressed as the mean ± S. E. in triplicate tests. **: p < 0.01, ***: p < 0.001 vs. control by ANOVA and Dunnett's multiple comparison test. Non: AGS cells, Control: AGS cells + H. pylori,SHHT: AGS cells + H. pylori + Sohamhyungtang, CRTA: AGS cells + H. pylori + Total alkaloid of rhizome of Coptidis Rhizoma, BRBR: AGS cells + H. pylori + Berberine. The effects of SHHT, CRTA and BRBR itself on the AGS cells, they showed no difference compared with AGS cells (data was not shown).


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