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JKM > Volume 40(2); 2019 > Article
Lee and Baik: A Comparative Study on the Effects of Pinellia ternata, Zingiber officinale and Sobanhatang on Reflux Esophagitis

Abstract

Objectives

This study was carried out to observe and compare the effects of Pinellia ternata, Zingiber officinale and Sobanhatang on the reflux esophagitis induced by gastric fundus and pylorus ligation in mice with esomeprazole.

Methods

Antioxidant effects were measured by DPPH radical scavenging activity at four different concentration of 0.125, 0.25, 0.5 and 1.0mg/102μℓ. Zingiber officinale water extract(ZE), Pinellia ternata water extract(PE) and Sobanhatang water extract(SBE) and esomeprazole were treated orally for 14 days before gatric fundus and pylorus ligation. In the histochemistry, changes in suface mucous cells, muscle tissue and connective tissue in gastro esophageal junction(GEJ) and mast cell on the esophageal mucosa were observed. The change of Hemo oxygenase(HO)-1, ghrelin, gastrin and substance P in gastric body tissue were measured by immunohistochemistry.

Results

DPPH radical scavenging activity exhibited concentration dependently increases in ZE, PE, SBE. ZE was significantly higher at all concentrations than PE. The gastric surface mucous cells were more in the treated group than in the reflux esophagitis elicited group(GE) in the order of PE, SBE, ZE and esomeprazole treateded group(PT, SBT, ZT, ET). Lower esophageal sphincter muscle damage and intercellular space in the GEJ were less in the treated group than GE. In the esophageal mucosa, the mast cell distribution and the migration of inflammatory cells were lower in the treateded troup than GE in order to ZT, SBT, PT and ET. The antioxidative enzyme, HO-1 was more in the order of ZT, SBT, control group, PT, ET than in GE. ZT was significantly higher than the other groups and SBT was significantly higher than ET. Ghrelin was found to be higher in ZT, ET, SBT and PT than in GE, and ZT was significantly higher than all other groups except ET. Gastrin showed the highest positivity in GE, and was lower in the order of ET, ZT, SBT, PT, and control group. Substance P was the highest in GE, and was lower in the order of ET, ZT, SBT, PT and control group, and PT were significantly lower than ET.

Conclusion

ZT, PT and SBT showed superior antioxidative, anti-inflammatory and mucosal protective effects on mouse reflux esophagitis as compared with ET. In particular, ZE was more effective in antioxidant and gastric motility enhancement, while PE was more effective in mucosal protection and anti-inflammatory effects. Sobanhatang is expected to be effective treatment because it has advantages of both drugs and reduces toxicity.

DPPH radical scavenging activity (%)=(1-absorbanceofsampleabsorbanceofblank)×100

Fig. 1
Experimental design and reflux esophagitis animal model
HFD, high fat diet; ZE, Zingiber officinale water extract; PE, Pinellia ternata water extract; SBE, Sobanhatang water extract.
jkm-40-2-17f1.gif
Fig. 2
The GC/MS of Component in water extracts of Zingiber officinale, Pinellia ternata, and Sobanhatang
jkm-40-2-17f2.gif
Fig. 3
The free radical scavenging activity of water extracts of Pinellia ternata, Zingiber officinale and Sobanhatang
The anti-oxidant ability of ZE and SBE is dose-dependantly increased and the estimation of SBE is lower compared with ZE. ♦, Pinellia ternata water extract(PE); ■, Zingiber officinale water extract(ZE); ▲, Sobanhatang water extract(SBE); ●, Ascorbic acid as compared anti-oxidant.
jkm-40-2-17f3.gif
Fig. 4
The hemo oxygenase-1 positive reaction in gastric mucosa with reflux esophagitis
Ctrl, no reflux esophagitis elicited group; GE, reflux esophagitis elicited group; ET, esomeprazole treated group before reflux esophagitis elicitation; ZT, Zingiber officinale water extract treated group before reflux esophagitis elicitation; PT, Pinellia ternata water extract treated group before reflux esophagitis elicitation; SBT, Sobanhatang water extract treated group before reflux esophagitis elicitation; Bar size, 100μm; arrow, HO-1 positive reaction; ASM, apical surface of mucosa, LP, lamina propria.
jkm-40-2-17f4.gif
Fig. 5
The ghrelin positive reaction in gastric mucosa with reflux esophagitis
Bar size, 100μm; arrow, ghrelin positive reaction; ASM, apical surface of mucosa, LP, lamina propria.
jkm-40-2-17f5.gif
Fig. 6
The gastrin positive reaction in gastric mucosa with reflux esophagitis
Bar size, 100μm; arrow, gastrin positive reaction; ASM, apical surface of mucosa, LP, lamina propria.
jkm-40-2-17f6.gif
Fig. 7
The substance P positive reaction in gastric mucosa with reflux esophagitis
Bar size, 100μm; arrow, gastrin positive reaction; ASM, apical surface of mucosa, LP, lamina propria
jkm-40-2-17f7.gif
Fig. 8
The protective effect of damaged mucosal barrier by Pinellia ternata water extract, Sobanhatang water extract, Zingiber officinale water extract and esomeprazole
The PAS positive reaction (arrow) in mice pre-treateded with PE, SBE, ZE and esomeprazole noticeably maintain (PAS stain; Bar size, 50μm).
ASM, apical surface of mucosa; LU, lumen.
jkm-40-2-17f8.gif
Fig. 9
The mitigative effect of muscular damage & mucosal inflammation by Pinellia ternata water extract, Sobanhatang water extract, Zingiber officinale water extract and esomeprazole
A photo Bar size, 200μm; B photo Bar size, 50μm, GEJ, gastroesophageal junction; LES, lower esophageal sphincter; ES, esophagus; SM, submucosa; LU, lumen.
jkm-40-2-17f9.gif
Table 1
The Amount and Composition of Sobanhatang
Herbs Scientific Name Amount (g)
生薑 Zingiberis Rhizoma Crudus Zingiber officinale Roscoe 11.25
半夏 Pinellia Rhizoma Pinellia ternata Breitenbach 15.00

Total Amount 26.25
Table 2
The GC/MS of Component in Zingiber officinale water extract
NO RT(min) Area(%) Compounds
1 13.430 44.27 Zingerone
2 19.230 7.84 2-Heptanone
3 19.850 24.57 6-shogaol
4 20.202 23.32 Gingerol

RT, retention time.

Table 3
The GC/MS of Component in Pinellia ternata water extract
NO RT(min) Area(%) Compounds
1 6.395 68.16 Epoxyethane
2 8.921 31.84 Benzaldehyde
Table 4
The GC/MS of Component in Sobanhatang water extract
NO RT(min) Area(%) Compounds
1 6.423 12.52 Cyclopentasiloxane
2 8.921 13.07 Benzaldehyde
3 11.066 8.33 Isobutylamine
4 13.430 66.09 Zingerone
Table 5
DPPH Radical Scavenging Activity of Water Extracts of Pinellia ternata, Zingiber officinale and Sobanhatang Scavenging activity (%)
Concentration PE (mean±SD) ZE (mean±SD) SBE (mean±SD) p value*
1.0 mg 18.89±1.81b 66.49±2.26a 30.31±1.76a,b 0.000
0.5 mg 14.38±1.31b 46.44±3.10a 23.88±1.64a,b 0.000
0.25 mg 9.49±1.06b 29.68±3.53a 15.92±1.08a,b 0.000
0.125 mg 5.18±1.28b 16.05±2.83a 9.61±1.24a,b 0.000

PE, Pinellia ternata water extract; ZE, Zingiber officinaele water extract; SBE, Sobanhatang water extract;

a,b a and b are groups separated by Bonferroni correction method, and represent the same group, respectively.

* Statical significance was determined by Kruskal-Wallis test.

Table 6
The Image Analysis for the Hemo Oxygenase-1 Positive Reaction in Gastric Mucosa
Objective Group
Ctrl (mean±SD) GE (mean±SD) ET (mean±SD) ZT (mean±SD) PT (mean±SD) SBT (mean±SD) p value*
Hemo oxygenase-1 12225±2251b 4631±317d 8730±366c,d 43866±3023a 10082±1214b,c 22876±2025b 0.000

(image analysis for 20.000,000 particles/range of intensity : 80–100)

Ctrl, no reflux esophagitis elicited group; GE, reflux esophagitis elicited group; ET, esomeprazole treated group before reflux esophagitis elicitation; ZT, Zingiber officinale water extract treated group before reflux esophagitis elicitation; PT, Pinellia ternata water extract treated group before reflux esophagitis elicitation; SBT, Sobanhatang water extract treated group before reflux esophagitis elicitation;

a,b,c,d a, b, c and d are groups separated by Bonferroni correction method, and represent the same group, respectively.

* Statical significance was determined by Kruskal-Wallis test.

Table 7
The Image Analysis for the Ghrelin Positive Reaction in Gastric Mucosa
Objective Group
Ctrl (mean±SD) GE (mean±SD) ET (mean±SD) ZT (mean±SD) PT (mean±SD) SBT (mean±SD) p value*
Ghrelin 21888±1469c,d 18008±1077d 56342±2276a,b 59743±1704a 33323±1907b,c,d 47361±3862b,c 0.000

(image analysis for 20.000,000 particles/range of intensity : 80–100)

Table 8
The Image Analysis for the Gastrin Positive Reaction in Gastric Mucosa
Objective Group
Ctrl (mean±SD) GE (mean±SD) ET (mean±SD) ZT (mean±SD) PT (mean±SD) SBT (mean±SD) p value*
Gastrin 13897±1463d 97618±5148a 88260±1991a,b 78867±2963a,b,c 57045±1962c,d 66541±2351b,c,d 0.000

(image analysis for 20.000,000 particles/range of intensity : 80–100)

Table 9
The Image Analysis for the Substance P Positive Reaction in Gastric Mucosa
Objective Group
Ctrl (mean±SD) GE (mean±SD) ET (mean±SD) ZT (mean±SD) PT (mean±SD) SBT (mean±SD) p value*
Substance P 6498±535d 73238±3667a 53105±1580a,b 49622±2138a,b,c 15515±1332c,d 29171±1999b,c,d 0.000

(image analysis for 20.000,000 particles/range of intensity : 80–100)

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