Recommended Articles

(4) Journal of Pharmacopuncture, Vol. 17, No. 1, pp. 27–34, 2014

Single Intramuscular-dose Toxicity of Water soluble Carthmi-Flos Herbal Acupuncture (WCF) in Sprague-Dawley Rats

Hyung-geol Lee, Sungchul Kim, Da-jung Jung, Yoo-min Choi, Min-seop Sin, Seok-Woo Choi, Beom-yong Song, Jong-uk Kim, Seung-won Hong, Tae-han Yook*

*Corresponding Author’s Affiliation: Department of Acupuncture & Moxibustion Medicine, Woosuk University Hospital of Oriental Medicine, Jeonju, Korea. nasiss@naver.com

Abstract

Objectives: This experiment was conducted to examine the toxicity of Water soluble Carthmi-Flos herbal acupuncture (WCF) by administering a single intramuscular dose of WCF in 6-week-old, male and female Sprague-Dawley rats and to find the lethality dose for WCF.

Methods: The experiment was conducted at Biotoxtech according to Good Laboratory Practices under a request by the Korean Pharmacopuncture Institute. This experiment was performed based on the testing standards of “Toxicity Test Standards for Drugs” by the Ministry of Food and Drug Safety. Subjects were divided into 4 groups: 1 control group in which normal saline was administered and 3 test groups in which 0.1, 0.5 or 1.0 mL of WCF was administered; a single intramuscular dose was injected into 5 males and 5 females in each group. General symptoms and body weights were observed/ measured for 14 days after injection. At the end of the observation period, hematological and clinical chemistry tests were performed, followed by necropsy and histopathological examinations of the injected sections.

Results: No mortalities were observed in any group. Also, symptoms, body weight, hematology, clinical chemistry and necropsy were not affected. However, histopathological examination of the injected part in one female in the 1.0-mL group showed infiltration of mononuclear cells and a multi-nucleated giant cell around eosinophilic material.

Conclusion: Administration of single intramuscular doses of WCF in 3 groups of rats showed that the approximate lethal dose of WCF for all rats was in excess of 1.0 mL, as no mortalities were observed for injections up to and including 1.0 mL.

Keywords: Carthamus tinctorious L., Cathami Semen, Carthmi-Flos herbal acupuncture, water soluble Carthmi-Flos herbal acupuncture, pharmacopuncture, intramuscular toxicity test

(5) Journal of Pharmacopuncture, Vol. 17, No. 1, pp. 35–43, 2014

Single Dose Toxicity of Chukyu (spine-healing) Pharmacopuncture Injection in the Muscle of Rats

Hohyun Jeong, Seung-Hun Cho, Eun-Yong Lee, Seung-Deok Lee, Seong-Hun Ahn, Sungchul Kim*

*Corresponding Author’s Affiliation: Wonkwang Gwangju Oriental Medical Hospital, Gwangju, Korea. kscndl@hanmail.net

Abstract

Objectives: This study was performed to analyze the single dose toxicity of Chukyu (spine-healing) pharmacopuncture.

Methods: All experiments were conducted at the Biotoxtech, an institution authorized to perform non-clinical studies under the regulations of Good Laboratory Practice (GLP) regulations. Sprague-Dawley rats were chosen for the pilot study. Doses of Chukyu (spine-healing) pharmacopuncture, 0.1, 0.5 and 1.0 mL, were administered to the experimental groups, and a dose of normal saline solution, 1.0 mL, was administered to the control group. This study was conducted under the approval of the Institutional Animal Ethic Committee.

Results: No deaths or abnormalities occurred in any of the four groups. No significant changes in weight, hematological parameters or clinical chemistry between the control group and the experimental groups were observed. To check for abnormalities in organs and tissues, we used microscopy to examine representative histological sections of each specified organ; the results showed no significant differences in any of the organs or tissues except in one case, where interstitial infiltrating macrophages were found in one female rat in the 0.5-mL/animal experimental group.

Conclusion: The above findings suggest that treatment with Chukyu (spine-healing) pharmacopuncture is relatively safe. Further studies on this subject are needed to yield more concrete evidence.

Keywords: Chukyu (spine-healing) pharmacopuncture, toxicity test

(6) Journal of Pharmacopuncture, Vol. 17, No. 1, pp. 44–50, 2014

Isolation and Characterization of a 32-kDa Fibrinolytic Enzyme (FE-32kDa) from Gloydius blomhoffii siniticus Venom

Joung-Yoon Kim, Seung-Bae Lee, Ki Rok Kwon, Suk-Ho Choi*

*Corresponding Author’s Affiliation: Division of Animal Science and Biotechnology, Sangji University, Wonju, Korea. shchoi@sangji.ac.kr

Abstract

Objectives: This study was undertaken to isolate a fibrinolytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate its enzymatic characteristics and hemorrhagic activity as a potential pharmacopuncture agent.

Methods: The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were investigated using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined.

Results: The fibrinolytic enzyme with the molecular weight of 32kDa (FE-32kDa) from Gloydius blomhoffii siniticus showed a fibrin hydrolysis zone at the concentration of 0.2 mg/mL in the fibrin plate assay. The fibrin hydrolysis activity of the enzyme was inhibited completely by ethylenediaminetetraacetic acid (EDTA), ethyleneglycoltetraacetic acid (EGTA), and 1, 10- phenanthroline, thiothreitol and cysteine, and partially by phenylmethanesulfonylfluoride (PMSF). Metal ions such as Fe²⁺ and Hg²⁺ inhibited the fibrin hydrolysis completely, but Zn²⁺ enhanced it. FE-32kDa hydrolyzed α-chain but did not hydrolyze β-chain and γ-chain of fibrinogen. High-molecular-weight polypeptides of gelatin were hydrolyzed partially into low-molecular-weight polypeptides, but the extent of hydrolysis was limited. FE-32kDa induced hemorrhage beneath back skin of mice at the dose of 2 µg.

Conclusions: FE-32kDa is a α-fibrin(ogen)olytic metalloprotease that requires Zn²⁺ for fibrinolytic activity and causes hemorrhage, suggesting that the enzyme is not appropriate for use as a clinical pharmacopuncture.

Keywords: fibrinolytic enzyme, metalloprotease, Gloydius blomhoffii siniticus, snake venom