suppressive effect of berry phenolics on helicobacter pylori infection

1
Suppressive Effect of Berry Phenolics on Suppressive Effect of Berry Phenolics on Helicobacter Helicobacter pylori pylori Infection Infection RATIONALE & OBJECTIVES RATIONALE & OBJECTIVES Helicobacter pylori (H. pylori) is one of the major risk factors of gastric cancer. Thus, it is urgently necessary to develop a non-antibiotic alternative therapy to increase the eradication rates of H. pylori. Some literatures reported that cranberry may inhibit H. pylori adhesion to the human gastric mucus. However, the therapeutic possibility of other berries on H. pylori was rarely discussed. Thus, the aims of this experiment were to explore the impact of cranberry, blueberry, grape seed and mixed- berry extract on the growth, infectivity and pathogenicity of H. pylori in vitro. Agar disc diffusion method was used to compare the antimicrobial activities of the four berry extracts against H. pylori. For evaluating the inhibition of the four berry extracts on the adhesion of H. pylori, we first observed the adhesion of fluorescence- labeled H. pylori to the human gastric cancer cell line AGS under fluorescence microscope. Then, we used urease test to quantitate the H. pylori density adhesion to AGS cells. In addition, interleukin-8 secretion from AGS cells after H. pylori infection was measured by enzyme-linked immunosorbent assay. MATERIALS & METHODS MATERIALS & METHODS Ai-Piao Chang Ai-Piao Chang 1 Huey-Fang Shang Huey-Fang Shang 2 and Shih-Yi Huang and Shih-Yi Huang 1 * 1 School of Nutrition and Health Sciences, 2 School of Medicine, Taipei 1 School of Nutrition and Health Sciences, 2 School of Medicine, Taipei Medical University, Taipei, TAIWAN Medical University, Taipei, TAIWAN CONCLUSION CONCLUSION The results suggest that cranberry and other berry extracts may suppress the pathogenicity of H. pylori through the growth inhibition, reduction of H. pylori adhesion and H. pylori-induced IL-8 secretion from gastric cells. RESULTS & FINDINGS RESULTS & FINDINGS In results, inhibition zone assessment revealed that grape seed extract had the best inhibition effect on H. pylori in a dose-dependent manner. However, cranberry extract had no antimicrobial activity against H. pylori. (Fig. 1) Furthermore, the fluorescence microscopic observation showed that significant reduction of fluorescence- labeled H. pylori adhesion to AGS cells by berry extracts. The highest concentration of berry extracts against the adhesion of H. pylori to AGS cells and the percentage of adhesion inhibition determined by urease test were 20 mg/mL and 84% for cranberry extract, 5 mg/mL and 38% for blueberry extract, 10 mg/mL and 91% for grape seed extract, 20 mg/mL and 88% for GI extract, respectively. (Fig.2) In addition, the berry extract concentration and the percentage inhibition of H. pylori- induced IL-8 secretion from AGS cells were cranberry extract (5 mg/mL; 84%), blueberry extract (1.25 mg/mL; 97%), grape seed extract (0.075 mg/mL; 84%) and GI extract (0.15 mg/mL; 88%), respectively. (Fig.3) 0.7 3.5 7 0 5 10 15 20 25 30 C ranberry extract Blueberry extract G rape seed extract G Iextract P<0.001 P<0.001 B erry extract(m g) In hib ition zon e (m m) Figure 1:Inhibition zone comparison of all berry extracts. 5 10 20 0 25 50 75 100 * C ranberry extract(m g/mL) A iti-adhe sion activ ity (% of c ontro l) 1.25 2.5 5 0 25 50 75 100 * * * B lueberry extract (m g/mL) A iti-ad h esio n activity (% o f co n tro l) 2.5 5 10 0 25 50 75 100 * * * G rape seed extract(m g/mL) A iti-adhesion activity (% of control) 5 10 20 0 25 50 75 100 * * * GIextract(m g/m L) A iti-adhesion activity (% of control) Figure 2: Berry extracts inhibit H. pylori adhesion to AGS cells. *Significantly different from the control group (p<0.05). 0 100 200 300 400 500 600 * * * * IL -8 (p g/m L ) Figure 3: Berry extracts inhibit IL-8 secretion. ACKNOWLEDGEMENT ACKNOWLEDGEMENT The work was supported by the following grant to Dr S.-Y. Huang: SKH-TMU-98-02 from the Shin-Kong Wu Ho-Su Memorial Hospital and Taipei Medical University in Taiwan.

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RATIONALE & OBJECTIVES. MATERIALS & METHODS. RESULTS & FINDINGS. Figure 2: Berry extracts inhibit H. pylori adhesion to AGS cells . * Significantly different from the control group (p

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Page 1: Suppressive Effect of Berry Phenolics on  Helicobacter pylori  Infection

Suppressive Effect of Berry Phenolics on Suppressive Effect of Berry Phenolics on Helicobacter pyloriHelicobacter pylori InfectionInfection

RATIONALE & OBJECTIVESRATIONALE & OBJECTIVES

Helicobacter pylori (H. pylori) is one of the major risk factors of gastric cancer. Thus, it is urgently necessary to develop a non-antibiotic alternative therapy to increase the eradication rates of H. pylori. Some literatures reported that cranberry may inhibit H. pylori adhesion to the human gastric mucus. However, the therapeutic possibility of other berries on H. pylori was rarely discussed. Thus, the aims of this experiment were to explore the impact of cranberry, blueberry, grape seed and mixed-berry extract on the growth, infectivity and pathogenicity of H. pylori in vitro.

Agar disc diffusion method was used to compare the antimicrobial activities of the four berry extracts against H. pylori. For evaluating the inhibition of the four berry extracts on the adhesion of H. pylori, we first observed the adhesion of fluorescence-labeled H. pylori to the human gastric cancer cell line AGS under fluorescence microscope. Then, we used urease test to quantitate the H. pylori density adhesion to AGS cells. In addition, interleukin-8 secretion from AGS cells after H. pylori infection was measured by enzyme-linked immunosorbent assay.

MATERIALS & METHODSMATERIALS & METHODS

Ai-Piao ChangAi-Piao Chang11 Huey-Fang Shang Huey-Fang Shang22 and Shih-Yi Huang and Shih-Yi Huang11**1 School of Nutrition and Health Sciences, 2 School of Medicine, Taipei Medical 1 School of Nutrition and Health Sciences, 2 School of Medicine, Taipei Medical

University, Taipei, TAIWANUniversity, Taipei, TAIWAN

CONCLUSIONCONCLUSION

The results suggest that cranberry and other berry extracts may suppress the pathogenicity of H. pylori through the growth inhibition, reduction of H. pylori adhesion and H. pylori-induced IL-8 secretion from gastric cells.

RESULTS & FINDINGSRESULTS & FINDINGS In results, inhibition zone assessment revealed that grape seed extract had the best inhibition effect on H. pylori in a dose-dependent manner. However, cranberry extract had no antimicrobial activity against H. pylori. (Fig. 1) Furthermore, the fluorescence microscopic observation showed that significant reduction of fluorescence-labeled H. pylori adhesion to AGS cells by berry extracts.

The highest concentration of berry extracts against the adhesion of H. pylori to AGS cells and the percentage of adhesion inhibition determined by urease test were 20 mg/mL and 84% for cranberry extract, 5 mg/mL and 38% for blueberry extract, 10 mg/mL and 91% for grape seed extract, 20 mg/mL and 88% for GI extract, respectively. (Fig.2) In addition, the berry extract concentration and the percentage inhibition of H. pylori-induced IL-8 secretion from AGS cells were cranberry extract (5 mg/mL; 84%), blueberry extract (1.25 mg/mL; 97%), grape seed extract (0.075 mg/mL; 84%) and GI extract (0.15 mg/mL; 88%), respectively. (Fig.3)

0.7 3.5 70

5

10

15

20

25

30

Cranberry extractBlueberry extractGrape seed extractGI extract

P<0.001P<0.001

Berry extract (mg)

Inh

ibit

ion

zo

ne

(mm

)

Figure 1:Inhibition zone comparison of all berry extracts.

5 10 200

25

50

75

100

*

Cranberry extract (mg/mL)

Ait

i-ad

hesi

on a

cti

vit

y(%

of

co

ntr

ol)

1.25 2.5 50

25

50

75

100

** *

Blueberry extract (mg/mL)

Ait

i-ad

hesi

on a

ctiv

ity

(% o

f co

ntro

l)

2.5 5 100

25

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* **

Grape seed extract (mg/mL)

Ait

i-ad

hesi

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ctiv

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(% o

f co

ntro

l)

5 10 200

25

50

75

100

* * *

GI extract (mg/mL)

Ait

i-ad

hesi

on a

ctiv

ity

(% o

f co

ntro

l)

Figure 2: Berry extracts inhibit H. pylori adhesion to AGS cells. *Significantly different from the control group (p<0.05).

0

100

200

300

400

500

600

**

* *

IL-8

(p

g/m

L)

Figure 3: Berry extracts inhibit IL-8 secretion.

ACKNOWLEDGEMENTACKNOWLEDGEMENT

The work was supported by the following grant to Dr S.-Y. Huang: SKH-TMU-98-02 from the Shin-Kong Wu Ho-Su Memorial Hospital and Taipei Medical University in Taiwan.