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Phytochemical Analysis and Anti-cancer Investigation of Boswellia Serrata Bioactive Constituents In Vitro

  • Ahmed, Hanaa H;Abd-Rabou, Ahmed A;Hassan, Amal Z;Kotob, Soheir E
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7179-7188
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    • 2015
  • Cancer is a major health obstacle around the world, with hepatocellular carcinoma (HCC) and colorectal cancer (CRC) as major causes of morbidity and mortality. Nowadays, there isgrowing interest in the therapeutic use of natural products for HCC and CRC, owing to the anticancer activity of their bioactive constituents. Boswellia serrata oleo gum resin has long been used in Ayurvedic and traditional Chinese medicine to alleviate a variety of health problems such as inflammatory and arthritic diseases. The current study aimed to identify and explore the in vitro anticancer effect of B. Serrata bioactive constituents on HepG2 and HCT 116 cell lines. Phytochemical analysis of volatile oils of B. Serrata oleo gum resin was carried out using gas chromatography-mass spectrometry (GC/MS). Oleo-gum-resin of B. Serrata was then successively extracted with petroleum ether (extract 1) and methanol (extract 2). Gas-liquid chromatography (GLC) analysis of the lipoidal matter was also performed. In addition, a methanol extract of B. Serrata oleo gum resin was phytochemically studied using column chromatography (CC) and thin layer chromatography (TLC) to obtain four fractions (I, II, III and IV). Sephadex columns were used to isolate ${\beta}$-boswellic acid and identification of the pure compound was done using UV, mass spectra, $^1H$ NMR and $^{13}C$ NMR analysis. Total extracts, fractions and volatile oils of B. Serrata oleo-gum resin were subsequently applied to HCC cells (HepG2 cell line) and CRC cells (HCT 116 cell line) to assess their cytotoxic effects. GLC analysis of the lipoidal matter resulted in identification of tricosane (75.32%) as a major compound with the presence of cholesterol, stigmasterol and ${\beta}$-sitosterol. Twenty two fatty acids were identified of which saturated fatty acids represented 25.6% and unsaturated fatty acids 74.4% of the total saponifiable fraction. GC/MS analysis of three chromatographic fractions (I,II and III) of B. Serrata oleo gum resin revealed the presence of pent-2-ene-1,4-dione, 2-methyl- levulinic acid methyl ester, 3,5- dimethyl- 1-hexane, methyl-1-methylpentadecanoate, 1,1- dimethoxy cyclohexane, 1-methoxy-4-(1-propenyl)benzene and 17a-hydroxy-17a-cyano, preg-4-en-3-one. GC/MS analysis of volatile oils of B. Serrata oleo gum resin revealed the presence of sabinene (19.11%), terpinen-4-ol (14.64%) and terpinyl acetate (13.01%) as major constituents. The anti-cancer effect of two extracts (1 and 2) and four fractions (I, II, III and IV) as well as volatile oils of B. Serrata oleo gum resin on HepG2 and HCT 116 cell lines was investigated using SRB assay. Regarding HepG2 cell line, extracts 1 and 2 elicited the most pronounced cytotoxic activity with $IC_{50}$ values equal 1.58 and $5.82{\mu}g/mL$ at 48 h, respectively which were comparable to doxorubicin with an $IC_{50}$ equal $4.68{\mu}g/mL$ at 48 h. With respect to HCT 116 cells, extracts 1 and 2 exhibited the most obvious cytotoxic effect; with $IC_{50}$ values equal 0.12 and $6.59{\mu}g/mL$ at 48 h, respectively which were comparable to 5-fluorouracil with an $IC_{50}$ equal $3.43{\mu}g/mL$ at 48 h. In conclusion, total extracts, fractions and volatile oils of B. Serrata oleo gum resin proved their usefulness as cytotoxic mediators against HepG2 and HCT 116 cell lines with different potentiality (extracts > fractions > volatile oil). In the two studied cell lines the cytotoxic acivity of each of extract 1 and 2 was comparable to doxorubicin and 5-fluorouracil, respectively. Extensive in vivo research is warranted to explore the precise molecular mechanisms of these bioactive natural products in cytotoxicity against HCC and CRC cells.

Biological Activities of Isolated Icariin from Epimedium koreanum Nakai (삼지구엽초로부터 분리한 Icariin의 생리활성)

  • Kim, Seo-Jin;Park, Myoung-Su;Ding, Tian;Wang, Jun;Oh, Deog-Hwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.10
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    • pp.1397-1403
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    • 2011
  • Epimedium koreanum Nakai is a wild medicinal plant commonly consumed in South Korea due to its health beneficial effects. In the present study, the antioxidative, antimutagenic and immunological activities of E. koreanum Nakai extracts were investigated for their use in food. The yields of icariin compounds from the ethanol extract as well as the ethyl acetate, butanol, hexane, water, and chloroform fractions of E. koreanum were 27.9, 2.5, 1.7, 1.4, and 1.3 ${\mu}g/g$, respectively. The icariin components (295.5 ${\mu}g/g$) were collected from the ethyl acetate fraction by thin layer chromatography (TLC) and analyzed via high performance liquid chromatography (HPLC). The antioxidant activities of each fraction were as follows: ethyl acetate (49.0 ${\mu}g/mL$), butanol (59.2 ${\mu}g/mL$), hexane (119.8 ${\mu}g/mL$), water (122.0 ${\mu}g/mL$), and chloroform (138.5 ${\mu}g/mL$), based on $RC_{50}$ ${\mu}g/mL$. Icariin, isolated and identified as the main component, showed strong antioxidant activity with a $RC_{50}$ value of 15.3 ${\mu}g/mL$, which was higher than those of ascorbic acid (19.5 ${\mu}g/mL$) and ${\alpha}$-tocopherol (18.2 ${\mu}g/mL$). In an Ames test, none of the fractions produced mutagenic effects on Salmonella Typhimurium TA98 and TA100. In an immunomodulating activity test, the effects of E. koreanum Nakai on B cells (Rhamos) and T cells (Jurkat) were investigated. These results show that the growth and viability of B and T cells were increased by isolated icariin components for 1.27 and 1.28 fold, respectively. These results also provide preliminary data for the development of E. koreanum Nakai as an edible food material.

Lipid Components of Dried Laver(Cultured Porphyra tenera and Wild Porphyra suborbiculata) Produced at Wan-do in Korea (완도산 마른 참김(양식산)과 마른 둥근돌김(천연산)의 지방질 조성)

  • CHUNG Young-Hoon;LEE Eung-Ho;OH Kwang-Soo;CHA Yong-Jun;AHN Chang-Bum;LEE Tae-Hun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.18 no.5
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    • pp.433-438
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    • 1985
  • Lipids extracted from two kinds of sun-dried laver samples, wild Porphyra suborbiculata and cultured Porphyra tenera, produced at Wan-do in Korea were studied. Dried P. suborbiculata contained $0.8\%$ total lipid(TL) which consisted of $21.4\%$ neutral lipid(NL), $53.4\%$ glycolipid(GL) and $25.2\%$ phospholipid(PL), and dried P. tenera contained $1.2\%$ TL which consisted of $30.5\%$ NL, $50.3\%$ GL and $19.2\%$ PL. Among the NL of dried P. suborbiculate and P. tenera, free fatty acid ($41.4\%,\;39.0\%$), triglyceride($25.6\%,\;28.8\%$) and free sterol ($22.1\%,\;16.7\%$) were predominant. Digalactosyl diglyceride ($34.7\%,\;46.6\%$) and monogalactosyl diglyceride ($19.2\%,\;18.0\%$) were the major components among the GL. Sulfoquinovosyl digylceride ($4.2\%$) was also identified in P. tenera only. And main lipids in the PL of P. suborbiculata and P. tenera were phosphatidyl ethanolamine ($40.3\%,\;35.7\%$) and phosphatidyl choline ($28.6\%,\;30.7\%$) and followed by phosphatidyl serine($15.1\%,\;19.2\%$) and phosphatidyl inositol ($16.0\%,\;14.4\%$). The major fatty acids in the TL of the dried P. suborbiculata were 20:5 ($29.4\%$), 16:0 ($23.4\%$) and 20:4 ($13.0\%$), and those of the dried P. tenera were 20:5 ($36.7\%$), 16:0 ($16.2\%$), 16:1 ($10.7\%$) and 18:1 ($9.7\%$). The fatty acid composition of the both samples in the NL fraction were similar to the pattern in those of the TL. The abundant fatty acids in the PL of the both dried laver were 20:5, 16:0 and 18:1. In case of the GL fraction, the main fatty acids of the dried P. suborbiculata were 16:0, 20:5, 18:1 and 20:4, while those of the dried P. tenera were 20:5, 16:0 and 18:1.

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Lipid Components of Sea Squirt, Halocynthia roretzi, and Mideuduck, Styela clava (우렁쉥이 및 미더덕의 지방질성분)

  • Lee, Eung-Ho;Oh, Kwang-Soo;Lee, Tae-Hun;Ahn, Chang-Bum;Chung, Young-Hun;Kim, Kyung-Sam
    • Korean Journal of Food Science and Technology
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    • v.17 no.4
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    • pp.289-294
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    • 1985
  • Total lipid (TL) content of sea squirt (Ureungsweng-ee), Halocynthia roretzi, and Mideuduck, Styela clava, were 2.0%, 2.1%, respectively. Reviewing the composition of each lipid fraction in total lipids of sea squirt and Mideuduck, it was found that contents of neutral lipids (NL) (36.6%, 36.3%) and phospholipids (PL) (46.2%, 44.5%) were high, while that of glycolipids (GL) (17.2%, 19.2%) was low. The NL of sea squirt and Mideuduck were mainly consisted of triglyceride (49.0%, 59.6%) and free sterol (25.8%, 22.0%), and followed by diglyceride (9.4%, 7.7%), monoglyceride (6.0%, 4.2%), free fatty acid (4.6%, 1.9%) and esterified sterol and hydrocarbon (5.2%, 4.4%). And main lipids in PL were phosphatidylcholine (48.6%, 46.7%) and phosphatidylethanolamine (32.4%, 35.0%), and followed by phosphatidylinositol (9.8%, 7.0%), phosphatidylserine (5.7%, 5.8%) and an unknown substance (3.5%, 5.5%). Fatty acid composition was not significantly different among TL, NL, PL and GL contained in sea squirt and Mideuduck. The major fatty acids of TL in sea squirt and Mideuduck were eicosapentaenoic (21.3%, 18.3%), docosahexaenoic (16.3%, 14.2%), palmitic (13.8, 16.3%) and oleic acid (8.5%, 7.0%), respectively. Fatty acid composition of PL and NL were similar to those of TL. In case of GL fraction the major fatty acids were gadoleic (15.7%, 14.7%), palmitic (13.5%, 14.7%), stearic (11.6%. 9.8%) and oleic acid (8.0%, 8.1%).

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Comparison of Carotenoid Pigments on Manchurian Trout, Brachymystax lenok and Masu Salmon, Oncorhynchus macrostomus in the Family Salmonidae (연어과에 속하는 열목어와 산천어의 Carotenoid 색소성분의 비교)

  • BAEK Sung-Han;HA Bong-Seuk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.2
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    • pp.278-287
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    • 1998
  • Carotenoids in integument of wild manchurian trout, Brachymystax lenok, and wild and cultured masu salmon Oncohynchus macrostomus, which are all the Korean native cold fresh water fish, were investigated by thin layer chromatography, column chromatography and HPLC. The total carotenoid contents of the wild manchurian trout were $3.72\;mg\%$ which is relatively higher compare to other species of salmonidae. The carotenoids were composed of $36.9\%$ zeaxanthin and $14.7\%$ $\beta-carotene$ as the major compounds, $7.8\%$ triol $7.3\%$ isocryptoxanthin, $5.7\%$ 4-hydroxy echinenone, $4.7\%$ lutein, $4.5\%$ salmoxanthin and $2.2\%$ astaxanthin as minor compounds, and other carotenoids such as canthaxanthin, tunaxanthin A, tunaxanthin B, tunaxanthin C, $\beta-cryptoxanthin$ and $\alpha-cryptoxanthin$ as minute carotenoids. Wild masu salmon contained more total carotenoids than cultured one and the contents were $0.82\;mg\%$ and $0.66\;mg\%$, respectively. The composition of the carotenoids from wild masu salmon were $20.7\%$ xeaxanthin, $17.0\%$ isocryptoxanthin and $15.8\%\;\beta-carotene$ as major compounds, and $6.2\%$ triol, $6.1\%$ 4-hydroxy echinenone, $6.1\%$ salmoxanthin, $5.9\%$ canthaxanthin, $5.8\%$ lutein, $4.9\%$ $\alpha-cryptoxanthin$ and $1.0\%$ astaxanthin as minor compounds. The composition of the carotenoids from cultured masu salmon were $19.7\%$ isocryptoxanthin, $18.0\%$ $\beta-carotene$ and $10.3\%$ zeaxanthin as the major compounds, and $8.9\%\;\beta-cryptoxanthin$, $8.5\%\;\alpha-cryptoxanthin$, $8.0\%$ lutein, $7.6\%$ canthaxanthin, $5.1\%$ triol and $2.0\%$ astaxanthin as minor carotenoids. Based on these data, wild masu salmon contained more zeaxanthin, salmoxanthin and 4-hydroxy echinenone while cultured masu salmon contained more $\alpha-cryptoxanthin$, indicating that carotenoid pigment of masu salmon depends on their living conditions. Unlike wild masu salmon, 4-hydroxy echinenone and salmoxanthin which are the characteristic carotenoids of salmons, were not found in the integument of cultured masu salmon. Unlike manchurian trout, both wild and cultured masu salmon did not contain tunaxanthin A, tunaxanthin B and tunaxanthin C.

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Lipid Composition of Oyster, Arkshell and Sea-mussel (굴, 피조개 및 진주담치의 지질조성에 관한 연구)

  • YOON Ho-Dong;BYUN Han-Seok;CHUN Seok-Jo;KIM Seon-Bong;PARK Yeung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.4
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    • pp.321-326
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    • 1986
  • Oyster (Crassostrea gigas), arkshell (Anadare(Scapharce) broughtonii) and sea-mussel (Mytilus edulis) were investigated as to their lipid classes. Lipid extracts from shellfishes were fractionated into neutral lipid (NL), glycolipid (GL) and phospho-lipid (PL) by column chromatography with silicic acid. The fatty acid compositions of their lipid classes and lipid fractions were determined by gas liquid chromatography (GLC). Total lipid contents of shellfishes were $3.5\%$ in the oyster, $1.4\%$ in the arkshell, $1.0\%$ in the sea-mussel. The major fatty acids of total lipids were palmitic acid, eicosapentaenoic acid and docosahexaenoic acid in the oyster and the sea-mussel, palmitic acid, oleic acid and eicosapentaenoic acid in the arkshell. The lipid composition of neutral lipid fractions in shellfishes was separated and identified as free sterol, free fatty acid, triglyceride, hydrocarbon and esterified sterol by TLC. Of these classes, triglyceride fraction was most abundant, amounting to 55.6, 77.7 and $60.4\%$ in the three samples mentioned above, respectively. The main fatty acids of glycolipid were palmitic acid, eicosaenoic acid and docosahexaenoic acid in oyster, myristic acid, palmitic acid and palmitoleic acid in the arkshell, docosahexaenoic acid, linolenic acid and palmitic acid in the sea-mussel. The major fatty acids of phospholipid were palmitic acid, eicosapentaenoic acid and docosahexaenoic acid in the oyster and sea-mussel, palmitic acid, eicosapentaenoic acid and erucic acid in the arkshell.

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Antioxidant, Physiological Activities, and Acetylcholinesterase Inhibitory Activity of Portulaca oleracea Extracts with Different Extraction Methods (추출방법에 따른 쇠비름의 항산화, 생리활성 및 Acetylcholinesterase 저해활성)

  • Kwon, Yu-Ri;Cho, Sung-Mook;Hwang, Seung-Pil;Kwon, Gi-Man;Kim, Jae-Won;Youn, Kwang-Sup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.3
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    • pp.389-396
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    • 2014
  • The physiological properties of 70% ethanol extracts from Portulaca oleracea with different extraction methods (reflux extraction, RE; autoclave extraction, AE; low temperature high pressure extraction, LTPE) were investigated. The freeze-dried powder yields of RE, AE, and LTPE were 33.78%, 30.80%, and 11.05%, respectively. The color values of L and b were higher in LTPE, and the chroma values were higher in AE and LTPE compared to RE. The total polyphenolics and proanthocyanidin contents in LTPE were significantly higher than in other extracts. The amount of substances related to flavonoids contents was highest in RE (4.30 mg/g), followed by AE (4.06 mg/g), and LTPE (4.00 mg/g). DPPH radical scavenging ability with a concentration of 500 mg% (w/v) were in the following order; LTPE (88.87%)> RE (83.84%)> AE (80.67%). Further, the reducing power, ABTS radical scavenging ability, and nitrite scavenging activity was observed in the same tendency as seen with the DPPH radical scavenging ability. However, the ferrous ion chelating activity of RE (85.45%) and AE (83.88%) was significantly higher than that of LTPE (75.60%). ${\alpha}$-Glucosidase inhibitory activities of RE and LTPE with a concentration of 100 mg% were significantly higher than AE. Xanthine oxidase, and acetylcholinesterase inhibitory activities of LTPE were higher than the other extracts. These results suggest that the extracts from Portulaca oleracea have the potential to act as functional materials, and components of Portulaca oleracea could be effective in the prevention of Alzheimer's disease, and may be used to develop various functional food products.

The effect of nutritional supply on clinical outcomes and nutritional status in critically ill patients receiving continuous renal replacement therapy (지속적 신대체요법을 받은 중환자에서 영양공급이 임상결과와 영양상태에 미치는 영향)

  • Kim, Ju Yeun;Kim, Ji-Myung;Kim, Yuri
    • Journal of Nutrition and Health
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    • v.48 no.3
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    • pp.211-220
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    • 2015
  • Purpose: This study was designed to investigate whether nutritional supply influences biochemical markers and clinical outcomes in patients who received continuous renal replacement therapy (CRRT) by evaluating adequacy of nutritional supply for patients. Methods: From January 2012 to December 2013, 239 adult patients who received CRRT in the intensive care unit for more than 3 days were included. General information from electronic medical records and nutritional status related biochemical data and clinical outcomes on the first day of CRRT and 2 weeks after CRRT were collected. Results: The rate of delivered energy and protein was 68.06% and 43.13% which was much lower than energy and protein supply based on their requirement. When the patients were divided into two groups according to 70% of energy received rate and 50% of protein received rate, the group with more than 70% of energy received rate showed significant decrease of length of hospital stay (p = 0.007), length of stay in intensive care unit (ICU) (p = 0.008), duration of CRRT (p < 0.001), and APACHE II score (p < 0.001) compared to less than 70% of energy received rate after adjusting for age. In addition, the group with more than 50% of protein received rate showed decreased mortality (p = 0.031), length of hospital stay (p = 0.008), length of ICU stay (p = 0.035), duration of CRRT (p < 0.001), and APACHE II score (p < 0.001) after adjusting for age. We found that the level of hematocrit (p = 0.006) was significantly improved in the group with more than 70% of energy received rate, and the level of TLC (p = 0.049), hematocrit (p = 0.041) was significantly improved in the group with more than 50% of protein received rate. We also found that energy delivery was negatively correlated with length of stay in ICU (p = 0.049) and positively correlated with level of calcium (p = 0.037). In addition, protein delivery was correlated with the levels of serum total protein (p = 0.021), serum albumin (p = 0.048), hematocrit (p = 0.009), and total cholesterol (p = 0.021) when dead patients were included, but was correlated with the levels of hematocrit (p = 0.034) and calcium (p = 0.024) when dead patients were excluded. Conclusion: Proper nutritional delivery may help patients' clinical outcomes for patients receiving CRRT. However, their actual intakes of energy and protein were not adequate for their requirements. Identification of patients with malnutrition is necessary and a multidisciplinary approach for systemic management is also required.

Purification and Biological Activity of Ecdysterone from Korean Achyranthes radix (韓國産 牛膝의 Ecdysterone 抽出과 그 生理活性에 관한 硏究)

  • Kim, Jeong-Il;Lee, Jae-Yong;Kim, Chun-Su;Park, Kwang-E.
    • Journal of Sericultural and Entomological Science
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    • v.25 no.1
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    • pp.1-20
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    • 1983
  • It has been known that the insect molting hormone and its analogues exist also in plant kingdom and their concentration has been found to be about 0.1~2.0% of dry matter, which is equivalent to $10^3{\sim}10^5$ times of those in insects. This study was carried out; 1) to isolate the phytoecdysones from Korean Achyranthes radix and characterize their physico-chemical properties. 2) to investigate the biological activity of this phytoecdysone on Bombyx mori larvae. The resuls were summarized as follows; 1. The extraction method of phytoecdysones was optimized by three consecutive reflux for 1hr using 200g of dried and milled radix per 1l methanol. 2. The purification from the crude extract was made by a series of steps such as precipitation of gum-type polymer with n-Butyl acetate, adsorption on technical grade silica and chromatography with neutral alumina. The conditions of each step were optimized and the resulting crude crystal was about 500mg per kg dry radix. 3. The crude crystal from the cultivated Achyranthes(Achyranthes japonia) contained ecdysterone (20-hydroxyecdysone) and inokosterone in the proportion of one to one. In order to separate these, a series of processes such as acetylation, separation by alumina column chromatography deacetylation by alcoholysis, deionization and crystallization were introduced and optimized 125mg of ecdysterone and 18mg of inokosterone per kg dry radix were thus obtained. 4. The wild Achyranthes (Achyranthes obtusifolia) radix was found to contain the ecdysterone only. A 285mg of ecdysterone was crystallized per kg dry radix. 5. Isolated ecdysterone, inodosterone and acetylated compounds were characterized by IR., UV., NMR spectroscopy, mp, TLC and densitometry. 6. Ligation experiment was undertaken to confirm the biological activity of the purified ecdysterone; the ecdysterone could induce larval-pupal metamorphosis in the ligated abdomen of 4th instar larvae injecting 0.5~1.0${\mu}g$. 7. By ecdysterone feeding experiment using artificial diet, it was elucidated that the critical time of feeding would be the first half of each instar resulting in increased weight of silk layer. 8. The ecdysterone was fed to 5th instar silkworm at the level of 1, 2, 3, 5ppm of dry feed of artificial diet containing 5% mulberry leaves for 72hrs. At 2ppm of the concentration. body weight and silk layer weight were arrived at maximum. But at higher concentrations body weight and silk layer weight decreased than the control group. At 2ppm of the concentration, body weight was increased by 12.5%. 9. Feeding 2ppm of ecdysterone at the later half of 5th instar, the duration of larvae was shortened.

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Antioxidant Activities and Cytoprotective Effects of Lonicera japonica Thunb. Extract and Fraction against Oxidative Stress (인동덩굴 추출물과 분획물의 항산화 활성 및 산화적 스트레스에 대한 세포 보호 효과)

  • Lee, Ye Seul;Yun, Mid Eum;Lee, Yun Ju;Park, Young Min;Lee, Sang Lae;Park, Soo Nam
    • Microbiology and Biotechnology Letters
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    • v.46 no.1
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    • pp.18-28
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    • 2018
  • In this study, the antioxidant activities and cytoprotective effects against oxidative stress of Lonicera japonica Thunb. 50% ethanol extract and ethyl acetate fraction were investigated. Using the 1,1-diphenyl-2-picrylhydrazyl assay, the free radical scavenging activity (FSC50) of L. japonica Thunb. 50% ethanol extract and ethyl acetate fraction was determined as 152.00 and $77.25{\mu}g/ml$, respectively. To measure the reactive oxygen species (ROS) scavenging activity, the total antioxidant capacity (OSC50) was determined by using a luminol-dependent chemiluminescence assay. The antioxidant activity of the ethyl acetate fraction ($0.33{\mu}g/ml$) was approximately four times stronger than that of the 50% ethanol extract ($1.12{\mu}g/ml$). The protective effect against $^1O_2$-induced cellular damage of human erythrocytes (${\tau}_{50}$) was 46.0 min at $10{\mu}g/ml$ of the 50% ethanol extract and 52.3 min at $1{\mu}g/ml$ of the ethyl acetate fraction. We also investigated the cytoprotective effects against oxidative stress induced by $H_2O_2$ and the intracellular ROS scavenging activity in response to UVB irradiation and found that the extract and fraction protected human skin cells from damage and reduced ROS. These results confirmed that L. japonica Thunb. was a valuable plant-derived natural antioxidant with potential for development as an antioxidative functional ingredient.