분자생물학적 방법에 의한 세포 독성 측정

정화진,김동욱,Chung Hwa-Jin,Kim Dong-Wook 한국작물학회 2004 Korean journal of crop science Vol.49 No.suppl1

황련해독탕 약침의 장기보존시험에 관한 연구

이진호 ( Jin Ho Lee ),하인혁 ( In Hyuk Ha ),김미령 ( Me Riong Kim ),정화진 ( Hwa Jin Chung ),이재웅 ( Jae Woong Lee ),김민정 ( Min Jeong Kim ),김은지 ( Eun Jee Kim ),이인희 ( In Hee Lee ) 한방재활의학과학회 2016 한방재활의학과학회지 Vol.26 No.2

Objectives We studied long-term preservation in stability of a mixed preparation of distilled and 70% alcohol extracted Hwangnyunhaedok-tang pharmacopuncture to establish standards for expiration date and quality control. Methods Three lots of consecutively prepared Hwangnyunhaedok-tang pharmacopuncture were each tested in triplicate to a total 5 tests at 3 month intervals over a period of 12 months for analysis of appearance, pH, specific gravity, index component content, endotoxins, microbial sterility, residual organic solvents, heavy metals, and pesticides. Items with no difference by elapsed time were tested at the initial and final timepoints, and data of items with potential difference by elapsed time were analyzed for trends to establish individual quality control standards. Results All tested items were stable over the study period, and therefore the expiration date was set as 12 months. pH quality control standards were set as 3.66∼5.69, and that of specific gravity as 0.802∼1.203, respectively. In index component content standards, berberine was set at 4.96∼8.98 μg/vial, baicalin at 6.47∼10.31 μg/vial, and geniposide at 116.03∼189.55 μg/vial, respectively. Standards for other items with no difference by elapsed time were set according to general Korean herbal medicine standards in the Korean Pharmacopoeia. Conclusions Manageable expiration date and quality control standards were established through long-term preservation testing of Hwangnyunhaedok-tang pharmacopuncture, furthering standardization of Korean medicine pharmacopuncture. (J Korean Med Rehab 2016;26(2):51-59)

신바로 약침의 SD 랫드와 비글견에 대한 단회 근육투여 독성시험

이진호 ( Jin Ho Lee ),정화진 ( Hwa Jin Chung ),이인희 ( In Hee Lee ),이재웅 ( Jae Woong Lee ),김은지 ( Eun Jee Kim ),김민정 ( Min Jeong Kim ) 한방재활의학과학회 2015 한방재활의학과학회지 Vol.25 No.3

Objectives To evaluate Shinbaro Pharmacopuncture safety through analysis of potential single-dose intramuscular toxicity of Sinbaro Pharmacopucture in SD rats and Beagle dogs. Methods Single-dose intramuscular toxicity of Shinbaro Pharmacopuncture was assessed in accordance with Korea Food and Drug Administration Guidelines for toxicity testing of Medicinal Products. The SD rats were treated intramuscularly with Shinbaro Pharmacopuncture at doses of 0, 4.6, 9.2, and 18.5 mg/kg, respectively. The Beagle dogs were treated intramuscularly with Shinbaro Pharmacopuncture at doses of 2.3, and 4.6 mg/kg, respectively, and after 3 days, the procedure was repeated a second time at doses of 0.6, and 1.2 mg/kg, respectively, for toxicity testing. Mortality, change in body weight, and necropsy findings were examined for the study period. Results There were no mortalities, general symptoms, or body weight changes in the SD rats. While pyelectasis of the left kidney was observed in a male rat in the 4.6 mg/kg administration group, natural occurrence is common, and does not appear to be related with the test substance. No mortalities were observed in the Beagle dogs. In assessment of general symptoms, a female dog in the 9.2 mg/kg group displayed body weight decrease due to leftover food, but the change in body weight was within the normal range seen at 6~7 months, and the necropsy findings were not significant. The toxicity of the test substance appears to be minimal. Conclusions The results suggest that the lethal dose 50 (LD50) and approximate lethaldose (ALD) value in single intramuscular administration of Shinbaro Pharmacopuncture in SD rats and Beagle dogs are higher than 18.5 mg/kg. (J Korean Med Rehab 2015;25(3):1-9)

두충의 건조법별 지표성분 및 항염증 활성 비교

도호정 ( Ho-jeong Do ),이진호 ( Jin-ho Lee ),하인혁 ( In-hyuk Ha ),정화진 ( Hwa-jin Chung ),이재웅 ( Jae-woong Lee ),김민정 ( Min-jeong Kim ),김은지 ( Eun-jee Kim ),이인희 ( In-hee Lee ) 대한한의학회 한방재활의학과학회 2017 한방재활의학과학회지 Vol.27 No.1

Objectives Eucommiae cortex is one of the frequently used herbs for musculoskeletal disorders, with well-known anti-inflammatory effect. Meanwhile, powdered form of herbal drugs is more advantageous than decoction form, in that storing and taking is more convenient. Thus, the authors aimed to investigate whether the drying method affects the index compound level and anti-inflammatory effect of eucommiae cortex water extract. Methods Eucommiae cortex was extracted in distilled water at 100℃ for 3 hours, filtered, and then evaporated under vacuum. One half of the sample was freeze-dried at -80℃. Another half of the sample was added with dextrin and then spray-dried at 180℃. To assess the possible change in index compound content, pinoresinol diglucoside was selected as the index marker. The content level of the index compound in various extract sample was quantified through high performance liquid chromatography. In addition, iNOS assay in LPS-induced RAW 264.7 cells was adopted to determine the anti-inflammatory effect of various extract samples. Furthermore, MTT assay was performed to confirm that the result of iNOS assay was not due to cytotoxicity. Results There was no significance difference in index compound content between extract samples obtained through two different drying methods. Anti-inflammatory activity of extract samples were similar at the matching concentration, regardless of the drying methods. Extract samples did not show any significant cytotoxicity. Conclusions Extract samples of eucommiae cortex were obtained through freeze-drying and spray-drying. Neither change in index compound content nor difference in anti-inflammatory activity was observed between drying methods. (J Korean Med Rehabil 2017;27(1):19-25)

염수초 포제법에 따른 두충의 항산화 및 항염증 활성 변화 비교연구

고원일 ( Wonil Koh ),이진호 ( Jinho Lee ),하인혁 ( In-hyuk Ha ),정화진 ( Hwa-jin Chung ),이인희 ( In-hee Lee ),이재웅 ( Jae-woong Lee ),김은지 ( Eun Jee Kim ),강병구 ( Byeong-gu Gang ),전세환 ( Se Hwan Jeon ),조용규 ( Yongkyu Cho 대한한의학회 한방재활의학과학회 2017 한방재활의학과학회지 Vol.27 No.2

Objectives The present study aimed to investigate the change in marker compounds, anti-oxidative and anti-inflammatory effects of salt-water processed Cortex Eucommiae. Methods To evaluate the influence of processing on anti-oxidant effect of Cortex Eucommiae, changes in total phenol, total flavonoid, 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging, 2,2`-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) free radical scavenging, and ferric reducing antioxidant power (FRAP) between processed and raw Cortex Eucommiae were assessed. In addition, nitrite assay was conducted to determine the influence of processing on anti-inflammatory effect of Cortex Eucommiae. Cell viability was also examined as to elucidate whether processing affects cytotoxicity of Cortex Eucommiae. Finally, high-performance liquid chromatography (HPLC) analysis was conducted to monitor changes in pinoresinol diglucoside amount of processed and raw Cortex Eucommiae. Results Salt-water processed Cortex Eucommiae showed higher total phenol and flavonoid amount, compared to raw Cortex Eucommiae. Furthermore, anti-oxidative activity of processed Cortex Eucommiae was improved as discovered in DPPH, ABTS, and FRAP assays. Anti-inflammatory effect of Cortex Eucommiae was also enhanced following salt-water processing, as evidenced in nitrite assay. HPLC analysis found that the amount of pinoresinol diglucoside, widely known as the marker compound of Cortex Eucommiae, increases through salt-water processing. All experiments were performed with non-toxic concentration of Cortex Eucommiae; processing did not affect the cytotoxicity of Cortex Eucommiae up to the currently adopted concentration. Conclusions The present results support that salt-water processing of Cortex Eucommiae is beneficial in terms of marker compound amount, anti-oxidative, and anti-inflammatory activities. Additional investigations are needed to standardize the processing method of Cortex Eucommiae. (J Korean Med Rehabil 2017;27(2):29-38)