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The interaction of serum albumin with ginsenoside Rh2 resulted in the downregulation of ginsenoside Rh2 cytotoxicity

  • Lin, Yingjia;Li, Yang;Song, Zhi-Guang;Zhu, Hongyan;Jin, Ying-Hua
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.330-338
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    • 2017
  • Background: Ginsenoside Rh2 (G-Rh2) is a ginseng saponin that is widely investigated because of its remarkable antitumor activity. However, the molecular mechanism by which (20S) G-Rh2 triggers its functions and how target animals avoid its cytotoxic action remains largely unknown. Methods: Phage display was used to screen the human targets of (20S) G-Rh2. Fluorescence spectroscopy and UV-visible absorption spectroscopy were used to confirm the interaction of candidate target proteins and (20S) G-Rh2. Molecular docking was utilized to calculate the estimated free energy of binding and to structurally visualize their interactions. MTT assay and immunoblotting were used to assess whether human serum albumin (HSA), bovine serum albumin (BSA), and bovine serum can reduce the cytotoxic activity of (20S) G-Rh2 in HepG2 cells. Results: In phage display, (20S) G-Rh2-beads and (20R) G-Rh2-beads were combined with numerous kinds of phages, and a total of 111 different human complementary DNAs (cDNA) were identified, including HSA which had the highest rate. The binding constant and number of binding site in the interaction between (20S)-Rh2 and HSA were $3.5{\times}10^5M^{-1}$ and 1, and those in the interaction between (20S) G-Rh2 and BSA were $1.4{\times}10^5M^{-1}$ and 1. The quenching mechanism is static quenching. HSA, BSA and bovine serum significantly reduced the proapoptotic effect of (20S) G-Rh2. Conclusion: HSA and BSA interact with (20S) G-Rh2. Serum inhibited the activity of (20S) G-Rh2 mainly due to the interaction between (20S) G-Rh2 and serum albumin (SA). This study proposes that HSA may enhance (20S) G-Rh2 water solubility, and thus might be used as nanoparticles in the (20S) G-Rh2 delivery process.

Differential Role of protein Kinase C in Ginsenoside $Rh_2$ - induced Apoptosis in SK-N-BE(2) and C6Bu-1 Cells

  • Young Sook Kim;Sun
    • Proceedings of the Ginseng society Conference
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    • pp.244-252
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    • 1998
  • Ginsenoside Rh, (G-Rh,) from Panax ginseng induced morphological features of apoptosis and DNA fragmentation as a biochemical marker of apoptosis confirmed by TUNEL reaction and agarose gel electrophoresis in human neuroblastoma SK-N-BE(2) and rat glioma C6Bu-1 cells During apoptosis by G-Rh2, protein kinase C (PKC) isoforms were analysed by immunoblotting. In SK-N-BE(2) cells, the levels of a, p and ${\gamma}$ subtypes were increased by undergoing apoptosis, while PKC e isoform increased early in treatment (3 h and 6 h). In addition, PKC s isoform gradually decreased during apoptosis by G-Rh2 and PKC $\theta$ isoform was detected in neither untreated- nor G-Rh1-treated SK-N-BE(2) cells (data not shown). However, no significant changes in the level of S and s isoforms were observed in C6Bu-1 cells undergoing apoptosis by G-Rh2. These results suggest that PKC subtypes may play differential roles in apoptotic signal pathways and their roles can be cell type-specific in apoptosis induced by G-Rh2.

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Effect of Molecular Weight of Chitosan on Flocculation of Suspended Solids in Rhodamine 6G-Sodium Dodecyl Sulfate System (키토산 분자량에 따른 Rhodamine 6G-Sodium Dodecyl Sulfate계의 응집효과에 관한 연구)

  • Kim, Sung Hyun;Nah, Jae Woon;Oh, Jeong Sun;Song, Ki Dong
    • Journal of the Korean Chemical Society
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    • v.42 no.6
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    • pp.672-678
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    • 1998
  • Chitin was extracted from crab shell of Portuns triberculatus and deacethylated to yield chitosan with various molecular weights. The absorption and the fluorescence spectra of Rhodamine 6G(Rh 6G)-sodium dodecyl sulfate(SDS) and Rh 6G-chitosan systems were obtained. From the spectra, we observed that the absorption and the fluorescence intensity of Rh 6G-SDS system decreased when S/D(the concentration of SDS to that of Rh 6G ratio) was below or at 32, while they increased when S/D was above 32. From the suspended solid(SS) removal rate and the transmittance of Rh 6G-SDS-chitosan system, we found that when S/D ratio was 32 its flocculating behaviour was much stronger than Rh 6G-SDS system. As the concentration and the molecular weight of chitosan increased, we also found that S/D range was extended from 32 to 100. With increasing the molecular weight of chitosan, the SS removal rate increased around pH 2~9 but decreased remarkably at pH>12.

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Activin A/BMP2 Chimera (AB204) Exhibits Better Spinal Bone Fusion Properties than rhBMP2

  • Ryu, Dalsung;Yoon, Byung-Hak;Oh, Chang-Hyun;Kim, Moon-Hang;Kim, Ji-Yong;Yoon, Seung Hwan;Choe, Senyon
    • Journal of Korean Neurosurgical Society
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    • v.61 no.6
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    • pp.669-679
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    • 2018
  • Objective : To compare the spinal bone fusion properties of activin A/BMP2 chimera (AB204) with recombinant human bone morphogenetic protein (rhBMP2) using a rat posterolateral spinal fusion model. Methods : The study was designed to compare the effects and property at different dosages of AB204 and rhBMP2 on spinal bone fusion. Sixty-one male Sprague-Dawley rats underwent posterolateral lumbar spinal fusion using one of nine treatments during the study, that is, sham; osteon only; $3.0{\mu}g$, $6.0{\mu}g$, or $10.0{\mu}g$ of rhBMP2 with osteon; and $1.0{\mu}g$, $3.0{\mu}g$, $6.0{\mu}g$, or $10.0{\mu}g$ of AB204 with osteon. The effects and property on spinal bone fusion was calculated at 4 and 8 weeks after treatment using the scores of physical palpation, simple radiograph, micro-computed tomography, and immunohistochemistry. Results : Bone fusion scores were significantly higher for $10.0{\mu}g$ AB204 and $10.0{\mu}g$ rhBMP2 than for osteon only or $1.0{\mu}g$ AB204. AB204 exhibited more prolonged osteoblastic activity than rhBMP2. Bone fusion properties of AB204 were similar with the properties of rhBMP2 at doses of 6.0 and $10.0{\mu}g$, but, the properties of AB204 at doses of $3.0{\mu}g$ exhibited better than the properties of rhBMP2 at doses of $3.0{\mu}g$. Conclusion : AB204 chimeras could to be more potent for treating spinal bone fusion than rhBMP2 substitutes with increased osteoblastic activity for over a longer period.

Ginsenoside Rh2 inhibits proliferation of human promyelocytic HL-60 leukemia cells via $G_0/G_1$ phase arrest and induction of differentiation

  • Cho, Seoung-Hee;Kim, Dong-Hyun;Lee, Kyung-Tae
    • Proceedings of the Ginseng society Conference
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    • pp.3-12
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    • 2006
  • 1 The present work was performed to investigate the effects of ginsenoside Rh2 on proliferation, cell cycle-regulation and differentiation of human leukemia HL-60 cells as well as the underlying mechanisms for these effects. 2 Ginsenoside Rh2 potently inhibited the proliferation of HL-60 cells in both a dose- and time-dependent manner with an $IC_{50}$, $20{\mu}M$. 3 DNA flow-cytometry indicated that ginsenoside Rh2 markedly induced a $G_1$ phase arrest of HL-60 cells. 4 Among the $G_1$ phase cell cycle-related proteins, the levels of cyclin-dependent kinase(CDK)4, 6 and cyclin D1, cyclin D2, cyclin D3 were reduced by ginsenoside Rh2, whereas the steadystate levels of CDK2 and cyclin E were unaffected. 5 The protein levels of a CDK inhibitor p16, $p21^{CIP1/WAF1}$ and $p27^{KIP1}$ were markedly increased by ginsenoside Rh2. 6 Ginsenoside Rh2 markedly enhanced the binding of $p21^{CIP1/WAF1}$ and $p27^{KIP1}$ with CDK2 and CDK6, resulting in the reduced activity of both kinases and the hypophosphorylation of Rb protein. 7 We furthermore suggest that ginsenoside Rh2 is a potent inducer of the differentiation of HL-60 cells, based on observations such as a reduction of the nitroblue tetrazolium level, an increase in the esterase activities and phagocytic activity, morphology changes, and the expression of CD11b, CD14, CD64 and CD66b surface antigens. 8 In conclusion, the onset of ginsenoside Rh2-induced the $G_0/G_1$ arrest of HL-60 cells prior to the differentiation is linked to a sharp up-regulation of the $p21^{CIP1/WAF1}$ level and a decrease in the CDK2, CDK4 and CDK6 activities. This is the first report demonstrating that ginsenoside Rh2 potently inhibits the proliferation of human promyelocytic HL-60 cells via the $G_1$ phase cell cycle arrest and differentiation induction.

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Production of the Rare Ginsenoside Rh2-MIX (20(S)-Rh2, 20(R)-Rh2, Rk2, and Rh3) by Enzymatic Conversion Combined with Acid Treatment and Evaluation of Its Anti-Cancer Activity

  • Song, Bong-Kyu;Kim, Kyeng Min;Choi, Kang-Duk;Im, Wan-Taek
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1233-1241
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    • 2017
  • The ginsenoside Rh2 has strong anti-cancer, anti-inflammatory, and anti-diabetic effects. However, the application of ginsenoside Rh2 is restricted because of the small amounts found in Korean white and red ginsengs. To enhance the production of ginsenoside Rh2-MIX (comprising 20(S)-Rh2, 20(R)-Rh2, Rk2, and Rh3 as a 10-g unit) with high specificity, yield, and purity, a new combination of enzymatic conversion using the commercial enzyme Viscozyme L followed by acid treatment was developed. Viscozyme L treatment at pH 5.0 and $50^{\circ}C$ was used initially to transform the major ginsenosides Rb1, Rb2, Rc, and Rd into ginsenoside F2, followed by acid-heat treatment using citric acid 2% (w/v) at pH 2.0 and $121^{\circ}C$ for 15 min. Scale-up production in a 10-L jar fermenter, using 60 g of the protopanaxadiol-type ginsenoside mixture from ginseng roots, produced 24 g of ginsenoside Rh2-MIX. Using 2 g of Rh2-MIX, 131 mg of 20(S)-Rh2, 58 mg of 20(R)-Rh2, 47 mg of Rk2, and 26 mg of Rh3 were obtained at over 98% chromatographic purity. Then, the anti-cancer effect of the four purified ginsenosides was investigated on B16F10, MDA-MB-231, and HuH-7 cell lines. As a result, these four rare ginsenosides markedly inhibited the growth of the cancer cell lines. These results suggested that rare ginsenoside Rh2-MIX could be exploited to prepare an anti-cancer supplement in the functional food and pharmaceutical industries.

Studies on the Ovarian Changes and Sex Hormone Concentrations in Holstein Cows with Ovarian Quiescence and Follicular Cystic Ovaries after Treatment with LH-RH and Gn-RH (LH-RH 및 Gn-RH 처리 무발정우와 난포낭종우의 혈장내 성호르몬 수준과 난소반응에 관한 연구)

  • 임영재;김상근
    • Korean Journal of Animal Reproduction
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    • v.13 no.2
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    • pp.79-84
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    • 1989
  • This study was carried out to investigate the effect of LH-RH and Gn-RH treatment in Holstein cows with ovarian quiescence and follicular cystic ovaries. The cows with ovarian quiescence and follicular cystic ovaries injected intramuscularly with 100$\mu\textrm{g}$, 200$\mu\textrm{g}$ and 400$\mu\textrm{g}$ of LH-RH and 200$\mu\textrm{g}$ and 400$\mu\textrm{g}$ of Gn-RH respectively. The cows was diagnosed by repeated rectal palpation. The plasma progesterone and estradiol-17$\beta$ concentrations were assayed by radioimmunoassay methods. The resutls of this study were summarized as follows : 1. Ovulations were induced after treatment of LH-RH and Gn-RH. The concentrations of progesterone reached small peak level at luteal phase and estradiol-17$\beta$ reached obvious peak level with the development and maturation of the follicle during the periods of degeneration of the corpus luteum, and normal ovarian cycle activity started subsequently. 2. The cows with ovarian quiescence and follicular cystic ovaries were induced ovulation at 38.9$\pm$5.3 hrs. after treatment of LH-RH in 66.7% cows and at 52.7$\pm$7.9 hrs after treatment of Gn-RH in 60.0% cows respectively. 3. The good ovarian responses were indicated in treatment with 200$\mu\textrm{g}$ to 400$\mu\textrm{g}$ of LH-RH than those treated with 100$\mu\textrm{g}$ in cows with ovarian quiescence, and did not show difference of ovarian responses between treatments with 200$\mu\textrm{g}$ to 400$\mu\textrm{g}$ of Gn-RH in cows with follicular cystic ovaries.

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Output Enhancement of Rhodamine 6G Dye Laser by Rhodamine 560 Energy Transfer Dye (Rhodamine 560을 이용한 rhodamine 6G 색소 레이저의 출력 증가)

  • 장원권;이민희
    • Korean Journal of Optics and Photonics
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    • v.5 no.2
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    • pp.266-271
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    • 1994
  • The output power and the energy of Rh-6G dye laser were enhanced by the mixture of Rh-560 dye whose fluorescence spectrum was coincident with the absorption spectrum of Rh-6G. The argon filled coaxial flashlamp used for pulsed pumping and argon laser for CW pumping. The concentration of Rh-6G dye was optimized in each pumping method before Rh-560 dye was mixed in Rh-6G dye solution. In the coaxial flash lamp pumped Rh-6G laser the output energy was increased about 30% when Rh-560 was mixed at 1% of Rh-6G concentration. In the case of argon laser pumping with multiline, the output power was increased 18% at the concentration of 2.5%. In the single line laser pumping, the output power was enhanced more efficiently. The power enhancements were 72% and 88% when the pumping wavelengths were 488 nm and 514.5 nm respectively. ively.

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Output Enhancement of Coaxial Flashlamp Pumped Rhodamine 6G Dye Laser by Energy Transfer (에너지 전환에 의한 동축 섬광관 펌핑 Rhodamine 6G 색소 레이저의 출력 증가)

  • 장우권
    • Korean Journal of Optics and Photonics
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    • v.4 no.2
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    • pp.195-199
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    • 1993
  • The output energy of Rh-6G dye laser was enhanced by the energy transfer in the mixture of Rh-6G and C-545. The laser was pumped by coaxial flashlamp filled argon gas. The optimum concentration of Rh-6G was $10_-4mol/l$ without mixing. The output energy was enhanced about 70 % at 0.4 % C-545 mixture with respect to the concentration of Rh-6G. The peak output power and the output energy were 27 kW and 50 mJ at the pumping energy of 346 J.

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Induced Ovulation in the Mandarin Fish, Siniperca scherzeri by Sex-Maturation Hormones (성성숙 호르몬 처리에 의한 쏘가리의 배란 유도)

  • 장선일;이완옥;이종윤;손송정
    • Journal of Aquaculture
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    • v.11 no.4
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    • pp.513-519
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    • 1998
  • Ovulation of maturing femal mandarin fish, Siniperca scherzeri was induced using single injection of human chorionic gonadotropin (HCG) or gonadotropin releasing hormone-analogue (GnRH-a), GnRH-a plus prostaglandin F2 (PG$F_2$) or GnRH-a plus pimozide. The response was evaluated by fertilization, embryo-formation and hatching rate after insemination. Those rates were generally higher in GnRH-a group than in HCG group. The higher hatching rat of above 89% was achived using a dosage of 5,000 IU/kg HCG plus 10 ${\mu}$g/kg GnRH-a, 10${\mu}$g/kg GnRH-a plus 500 ng/kg PGF2, and 10 ug/kg GnRH-a plus 1-5 mg/kg pimozide. Ovulation was induced in all female injected with sex-maturation hormones and stimulator, but blocked in female injected with HCG plus GnRH-a plus dopamine combination, and GnRH-a plus PGF2 plus indometacin combination. These results show that the mandarin fish in spawning period secrete a sex-mutruation assosiated hormones and gonadotropin-releasing -inhibiting factor(GRIF).

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