In vitro and in vivo Antibacterial Activities of Cinnamomum cassia Extracts Against Helicobacter pylori

Hyun-A Lee; Sunhwa Hong; Hong-Geun Oh; Sang-Ho Park; Youn-Chul Kim; Gil-Saeng Jeong; Okjin Kim

doi:10.5625/lar.2010.26.1.21

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Lee, Hong, Oh, Park, Kim, Jeong, and Kim: In vitro and in vivo Antibacterial Activities of Cinnamomum cassia Extracts Against Helicobacter pylori

Original Article

Lab. Anim. Res. 2010;26(1):21-29.

Published online: 17 December 2010

DOI: https://doi.org/10.5625/lar.2010.26.1.21

In vitro and in vivo Antibacterial Activities of Cinnamomum cassia Extracts Against Helicobacter pylori

Hyun-A Lee¹, Sunhwa Hong¹, Hong-Geun Oh², Sang-Ho Park³, Youn-Chul Kim⁴, Gil-Saeng Jeong⁵, Okjin Kim^1,^∗

¹Center for Animal Resources Development, Wonkwang University, Iksan, Korea

²Huvet Co. Ltd, Iksan, Korea

³Korea DNA Valley Co. Ltd, Iksan, Korea

⁴College of Pharmacy, Wonkwang University, Iksan, Korea

⁵Zoonosis Research Center, Wonkwang University, Iksan, Korea

^∗Corresponding author: Okjin Kim, Center for Animal Resources Development, Wonkwang University, 344-2 Shinyoung-Dong, Iksan, 570-749, Korea. TEL: +82-63-850-6668 FAX: +82-63-850-7308 E-mail: kimoj@wonkwang.ac.kr

Received 7 February 2010 Revised 2 March 2010 Accepted 12 March 2010

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

Infection with Helicobacter pylori (H. pylori) is strongly associated with duodenal and gastric ulcers. Substantial epidemiological data has revealed that high rates of H. pylori infection might be related to high rates of gastric cancer and gastric adenocarcinoma. In this study, a medicinal herbal plant, Cinnamomum cassia, was examined and screened for anti-H. pylori activity. Seventy percent ethanol was used for herbal extraction. For anti-H. pylori activity screening, inhibitory zone tests as an in vitro assay and in vivo study using a Mongolian gerbil (Meriones unguiculatus) model were performed. Also, the safety of herbal compounds was evaluated by animal study. As a result of inhibitory zone test, Cinnamomum cassia extract demonstrated strong anti-H. pylori activities. Also, as results of in vivo animal studies, Cinnamomum cassia demonstrated strong therapeutic effects against H. pylori infection according to the criteria of histological examination and rapid urease test. As results of the safety study, after 28 days treatment of the Cinnamomum cassia extract, the animals were not detected any grossly and histological changes. These results demonstrate that it can be successfully cured against H. pylori infection and protected from H. pylori-induced pathology with Cinnamomum cassia. It could be a promising native herb treatment for patients with gastric complaints including gastric ulcer caused by H. pylori.

Keywords: Cinnamomum cassia, Helicobacter pylori, antibacterial activity, Mongolian gerbil

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Figure 1.

The anti-H. pylori activities of Cinnamomum cassia extract using the paper disk diffusion bioassay. The diameter of clear zone reveals anti-H. pylori activities of the inoculated materiel. C. cassia (10⁰): Cinnamomum cassia extract 1.5 mg, C. cassia (10−1): Cinnamomum cassia extract 0.15 mg.

Figure 2.

Gross findings of the stomach in the study on therapeutic effects of H. pylori infection with Cinnamomum cassia extract. A: H. pylori+Cinnamomum cassia, B: H. pylori+ PBS (black arrow: ulcer lesions), C: PBS+Cinnamomum cassia, D: PBS+PBS.

Figure 3.

Histopathological findings of the stomach in the study on therapeutic effects of H. pylori infection with Cinnamomum cassia extract. A: H. pylori+Cinnamomum cassia, B: H. pylori+ PBS (Cellular degeneration and atrophy in the mucous layer), C: PBS+Cinnamomum cassia, D: PBS+PBS.

Table 1.

Anti-H. pylori activities of Cinnamomum cassia extract using the paper disk diffusion bioassay

Treatment		Clear zone (mm)
Treatment		6 hrs	12 hrs	24 hrs	48 hrs	60 hrs
DMSO		0	0	0	0	0
Gentamycin		5	10	8	8	7
Ganamycin		5	9	8	7	7
Enrofloxacin		10	15	13	12	9
1 Cinnamomum cassia	10⁰ (1.5 mg)	10	14	12	12	10
1 Cinnamomum cassia	10−1 (0.15 mg)	5	7	6	6	5

Table 2.

Results of body weight changes of anti-H. pylori activities of Cinnamomum cassia extract using in vivo study

Group	Treatment	Body weights (gram)
Group	Treatment	O w.p.i.^a	1 w.p.i.	2 w.p.i	3 w.p.i	4 w.p.i
I	H. pylori+Cinnamomum^b	135.5± 2.05	137.0± 2.55	141.0± 2.00^∗	144.5± 3.55^∗	146.0± 1.55^∗
II	H. pylori+PBS	135.5±4.00	135.0± 3.00	136.0± 2.55	136.5± 2.55	137.0± 1.55
III	PBS+Cinnamomum	135.0± 5.10	141.0± 4.25	145.0± 4.00^∗	149.5± 2.55^∗	153.0± 2.40^∗
IV	PBS+PBS	135.5± 4.15	139.0± 3.55	144.0± 3.15^∗	148.0± 2.30^∗	152.0± 2.00^∗

^a Weeks post inoculation. Treatment weeks with test materials after Helicobacter pylori infection.

^b Cinnamomum cassia treatment was continuously inoculated with 400 mg/kg dose during 4 weeks after Helicobacter pylori infection.

^∗ Significantly different from the positive control Group II (P<0.05).

Table 3.

Results of rapid urease test with gastric mucosal tissues after the study on therapeutic effects of H. pylori infection with Cinnamomum cassia extract

Group	Inoculation^a		N	Positive reaction (Positive percent) ^b	Therapeutic No. (Therapeutic percent)
Group	H. pylori	Treatment	N	Positive reaction (Positive percent) ^b	Therapeutic No. (Therapeutic percent)
I	Yes	Cinnamomum	10	0^∗ (0 %, CI ^c 0-25.9)	10^∗ (100 %, CI 74.1-100)
II	Yes	PBS	10	10 (100 %, CI 74.1-100)	0 (0 %, CI 0-25.9)
III	No	Cinnamomum	10	0^∗ (0 %, CI 0-25.9)	-
IV	No	PBS	10	0^∗ (0 %, CI 0-25.9)	-

^a Treatment was conducted daily during 4 weeks after H. pylori infection.

^b The positive percent revealed H. pylori colonization, which was observed as red color change from yellow color medium.

^c Incidence percentage (95% confidential interval) was calculated with MiniTab statistic software program.

^∗ Significantly different from the positive control Group II (P<0.05).

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