Full Text:   <1387>

CLC number: R96

On-line Access: 2010-01-06

Received: 2010-02-22

Revision Accepted: 2010-06-29

Crosschecked: 2010-12-15

Cited: 13

Clicked: 3663

Citations:  Bibtex RefMan EndNote GB/T7714

-   Go to

Article info.
1. Reference List
Open peer comments

Journal of Zhejiang University SCIENCE B 2011 Vol.12 No.1 P.47-54

http://doi.org/10.1631/jzus.B1000057


Eucommia ulmoides Oliv. antagonizes H2O2-induced rat osteoblastic MC3T3-E1 apoptosis by inhibiting expressions of caspases 3, 6, 7, and 9


Author(s):  Jun Lin, Yi-jing Fan, Christian Mehl, Jia-jun Zhu, Hong Chen, Ling-yan Jin, Jing-hong Xu, Hui-ming Wang

Affiliation(s):  Department of Stomatology, the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, China, Department of Prosthodontics, Propaedeutics and Dental Materials, School of Dentistry, Christian-Albrechts University, Kiel 24105, Germany, Department of Plastic Surgery, the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, China

Corresponding email(s):   wangmysm@yahoo.com.cn

Key Words:  Eucommia ulmoides Oliv., Duzhong, Oxidative damage, H2O2, MC3T3-E1 cells, Free radicals, Apoptosis


Jun Lin, Yi-jing Fan, Christian Mehl, Jia-jun Zhu, Hong Chen, Ling-yan Jin, Jing-hong Xu, Hui-ming Wang. Eucommia ulmoides Oliv. antagonizes H2O2-induced rat osteoblastic MC3T3-E1 apoptosis by inhibiting expressions of caspases 3, 6, 7, and 9[J]. Journal of Zhejiang University Science B, 2011, 12(1): 47-54.

@article{title="Eucommia ulmoides Oliv. antagonizes H2O2-induced rat osteoblastic MC3T3-E1 apoptosis by inhibiting expressions of caspases 3, 6, 7, and 9",
author="Jun Lin, Yi-jing Fan, Christian Mehl, Jia-jun Zhu, Hong Chen, Ling-yan Jin, Jing-hong Xu, Hui-ming Wang",
journal="Journal of Zhejiang University Science B",
volume="12",
number="1",
pages="47-54",
year="2011",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1000057"
}

%0 Journal Article
%T Eucommia ulmoides Oliv. antagonizes H2O2-induced rat osteoblastic MC3T3-E1 apoptosis by inhibiting expressions of caspases 3, 6, 7, and 9
%A Jun Lin
%A Yi-jing Fan
%A Christian Mehl
%A Jia-jun Zhu
%A Hong Chen
%A Ling-yan Jin
%A Jing-hong Xu
%A Hui-ming Wang
%J Journal of Zhejiang University SCIENCE B
%V 12
%N 1
%P 47-54
%@ 1673-1581
%D 2011
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1000057

TY - JOUR
T1 - Eucommia ulmoides Oliv. antagonizes H2O2-induced rat osteoblastic MC3T3-E1 apoptosis by inhibiting expressions of caspases 3, 6, 7, and 9
A1 - Jun Lin
A1 - Yi-jing Fan
A1 - Christian Mehl
A1 - Jia-jun Zhu
A1 - Hong Chen
A1 - Ling-yan Jin
A1 - Jing-hong Xu
A1 - Hui-ming Wang
J0 - Journal of Zhejiang University Science B
VL - 12
IS - 1
SP - 47
EP - 54
%@ 1673-1581
Y1 - 2011
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1000057


Abstract: 
Eucommia ulmoides Oliv. (EuO), also known as duzhong, native to China, has been reported to have antioxidative function, but its cellular mechanism is not fully examined yet. We investigated inhibitory effects of EuO leaf ethanol extracts on h2O2-induced apoptosis in rat osteoblastic MC3T3-E1 cells and underlying mechanisms. Locally-grown duzhong leaves were extracted with ethanol. MC3T3-E1 cells were treated with EuO (6.25, 12.5, 25, 50, and 100 µg/ml) for 24 h, and then h2O2 (800 µmol/L) for an additional 24 h. Cell survival rate, percentage of apoptosis, and expressions of caspases 3, 6, 7, and 9 were examined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, microscopic analysis, Western blotting, and reverse transcription polymerase chain reaction (RT-PCR). The final EuO leaf ethanol extract powder was detected to contain caffeotannic acid at 58 mg/g and geniposide at 3.45 mg/g by high performance liquid chromatography (HPLC). EuO remarkably restrained cell oxidative damage and increased cell survival rate in a dose-dependent manner: 0 µg/ml, 0.21; 6.25 µg/ml, 0. 28; 12.5 µg/ml, 0.31; 25 µg/ml, 0.48; 50 µg/ml, 0.54; and 100 µg/ml, 0.66 (P<0.05), with the half-effective concentration being around 25 µg/ml. MTT results were confirmed by microscopic analysis. Western blotting and RT-PCR analyses showed that the expressions of caspases 3, 6, 7, and 9 were significantly decreased in the EuO-treated cells compared with the control (EuO- and h2O2-free) (P<0.05), with the half-effective concentration of EuO ranging from 12.5 to 25 µg/ml. We conclude that the ethanol-extracted EuO leaf extracts promoted the growth of MC3T3-E1 cells, and suppressed the h2O2-induced apoptosis in a rat MC3T3-E1 osteogenic cell model, likely due to the inhibition of caspases’ activities. The results indicate that EuO is a potent antioxidant, which may contribute to its many cellular protective functions, including the promotion of bone growth.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1]Chen, G.H., Tian, Y.L., Yang, G.L., Wang, X.Y., Zhao, H.R., 2002. Determination of eliminating ratio of Chinese traditional medicine for hydroxyl radical by fluorescent spectrophotometry. Spectrosc. Spect. Anal., 22(4):634-636 (in Chinese).

[2]Chen, W.C., Luo, J., 2009. Extracting solution from Eucommia ulmoides Oliv. induces differentiation of goat bone marrow mesenchymal stem cells into osteoblasts and inhibits adipogenic differentiation. J. Clin. Rehab. Tissue Eng. Res., 13(10):1960-1964 (in Chinese).

[3]Chen, W.H., Lu, L.G., Zeng, M.D., Xu, Z.N., Liu, M., Mao, Y.M., Fang, J.Y., 2006. Effect of magnesium isoglycyrrhizinate on the proliferation and oxidative stress of rat hepatic stellate cells in vitro. Chin. J. Hepatol., 14(6):426-430 (in Chinese).

[4]Cho, E.J., Yokozawa, T., Rhyu, D.Y., Kim, S.C., Shibahara, N., Park, J.C., 2003. Study on the inhibitory effects of Korean medicinal plants and their main compounds on the 1,1-diphenyl-2-picrylhydrazyl radical. Phytomedicine, 10(6-7):544-551.

[5]Deyama, T., Nishibe, S., Nakazawa, Y., 2001. Constituents and pharmacological effects of Eucommia and Siberian ginseng. Acta Pharmacol. Sin., 22(12):1057-1070.

[6]Dimmeler, S., Haendeler, J., Nehls, M., Zeiher, A.M., 1997. Suppression of apoptosis by nitric oxide via inhibition of interleukin-1β-converting enzyme (ICE)-like and cysteine protease protein (CPP)-32-like proteases. J. Exp. Med., 185(4):601-607.

[7]Ha, H., Ho, J., Shin, S., Kim, H., Koo, S., Kim, I.H., Kim, C., 2003. Effects of Eucommiae Cortex on osteoblast-like cell proliferation and osteoclast inhibition. Arch. Pharm. Res., 26(11):929-936.

[8]Hansen, M.B., Nielsen, S.E., Berg, K., 1989. Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J. Immunol. Methods, 119(2):203-210.

[9]Kim, M.H., Chung, J., Yang, J.W., Chung, S.M., Kwag, N.H., Yoo, J.S., 2003. Hydrogen peroxide-induced cell death in a human retinal pigment epithelial cell line, ARPE-19. Korean J. Ophthalmol., 17(1):19-28.

[10]Lee, M.K., Cho, S.Y., Kim, D.J., Jang, J.Y., Shin, K.H., Park, S.A., Park, E.M., Lee, J.S., Choi, M.S., Lee, J.S., et al., 2005. Du-zhong (Eucommia ulmoides Oliv.) cortex water extract alters heme biosynthesis and erythrocyte antioxidant defense system in lead-administered rats. J. Med. Food, 8(1):86-92.

[11]Li, Y., Sato, T., Metori, K., Koike, K., Che, Q.M., Takahashi, S., 1998. The promoting effects of geniposidic acid and aucubin in Eucommia ulmoides Oliver leaves on collagen synthesis. Biol. Pharm. Bull., 21(12):1306-1310.

[12]Li, Y., Kamo, S., Metori, K., Koike, K., Che, Q.M., Takahashi, S., 2000a. The promoting effect of eucommiol from Eucommiae cortex on collagen synthesis. Biol. Pharm. Bull., 23(1):54-59.

[13]Li, Y., Metori, K., Koike, K., Kita, F., Che, Q.M., Sato, T., Shirai, W., Takahashi, S., 2000b. Granuloma maturation in the rat is advanced by the oral administration of Eucommia ulmoides Oliver leaf. Biol. Pharm. Bull., 23(1):60-65.

[14]Lin, J., Li, Y.Q., Li, W.H., Chen, H., 2008. Effects of calcium phosphate cement/Eucommia ulmoides extracts compound on cell proliferation and adhesion of MC3T3E1 osteoblasts in vitro. J. Zhejiang Univ. (Med. Sci.), 37(1):78-82 (in Chinese).

[15]Luo, J., Tian, C., Xu, J., Sun, Y., 2009. Studies on the antioxidant activity and phenolic compounds of enzyme-assisted water extracts from Du-zhong (Eucommia ulmoides Oliv.) leaves. J. Enzyme Inhib. Med. Chem., 24(6):1280-1287.

[16]Metori, K., Furutsu, M., Takahashi, S., 1997. The preventive effect of ginseng with Du-zhong leaf on protein metabolism in aging. Biol. Pharm. Bull., 20(3):237-242.

[17]Miyake, M., Arai, N., Ushio, S., Iwaki, K., Ikeda, M., Kurimoto, M., 2003. Promoting effect of kaempferol on the differentiation and mineralization of murine pre-osteoblastic cell line MC3T3-E1. Biosci. Biotechnol. Biochem., 67(6):1199-1205.

[18]Nakamura, T., Nakazawa, Y., Onizuka, S., Satoh, S., Chiba, A., Sekihashi, K., Miura, A., Yasugahira, N., Sasaki, Y.F., 1997. Antimutagenicity of Tochu tea (an aqueous extract of Eucommia ulmoides leaves): 1. The clastogen- suppressing effects of Tochu tea in CHO cells and mice. Mutat. Res., 388(1):7-20.

[19]Nishida, S., Kikuichi, S., Yoshioka, S., Tsubaki, M., Fujii, Y., Matsuda, H., Kubo, M., Irimajiri, K., 2003. Induction of apoptosis in HL-60 cells treated with medicinal herbs. Am. J. Chin. Med., 31(4):551-562.

[20]Ohmori, R., Iwamoto, T., Tago, M., Takeo, T., Unno, T., Itakura, H., Kondo, K., 2005. Antioxidant activity of various teas against free radicals and LDL oxidation. Lipids, 40(8):849-853.

[21]Prouillet, C., Maziere, J.C., Maziere, C., Wattel, A., Brazier, M., Kamel, S., 2004. Stimulatory effect of naturally occurring flavonols quercetin and kaempferol on alkaline phosphatase activity in MG-63 human osteoblasts through ERK and estrogen receptor pathway. Biochem. Pharmacol., 67(7):1307-1313.

[22]Yang, J., Kato, K., Noguchi, K., Dairaku, N., Koike, T., Iijima, K., Imatani, A., Sekine, H., Ohara, S., Sasano, H., Shimosegawa, T., 2003. Tochu (Eucommia ulmoides) leaf extract prevents ammonia and vitamin C deficiency induced gastric mucosal injury. Life Sci., 73(25):3245-3256.

[23]Yao, C.H., Tsai, H.M., Chen, Y.S., Liu, B.S., 2005. Fabrication and evaluation of a new composite composed of tricalcium phosphate, gelatin, and Chinese medicine as a bone substitute. J. Biomed. Mater. Res. B Appl. Biomater., 75(2):277-288.

[24]Yen, G.C., Hsieh, C.L., 2003. Inhibitory effect of Eucommia ulmoides Oliv. on oxidative DNA damage in lymphocytes induced by H2O2. Teratog. Carcinog. Mutagen, 23(s1):23-34.

Open peer comments: Debate/Discuss/Question/Opinion

<1>

Please provide your name, email address and a comment





Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - Journal of Zhejiang University-SCIENCE