The Detection of Osthole and Application of Cell Imaging Based on Nitrogen-Doped Carbon Dots

Ning Wang,Xuefang Yang,Haojiang Wang,Liping Xie,Weihua Jia,Wei Bian,Martin M. F. Choi 성균관대학교(자연과학캠퍼스) 성균나노과학기술원 2020 NANO Vol.15 No.03

A fluorescence probe has been synthesized for the detection of osthole using the nitrogen-doped carbon dots (NCDs) as shown in Fig. 1. The NCDs were fully characterized by transmission electron microscopy (TEM), X-ray diffraction (XRD), x-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR). Under the optimal experimental conditions, the NCDs fluorescence probe was highly selective and sensitive to osthole. The linear response range for osthole was 5.0–7 5μM with a detection limit of 38 nM. The mechanism of the interaction of osthole and NCDs was discussed. The fluorescence probe has been applied to the analysis of biological samples. The as-synthesized NCDs with high fluorescence intensity, low toxicity and good biocompatibility were applied to cell imaging.

Anti-inflammatory effects of osthole in peripheral blood mononuclear cells from Hanwoo (Bos taurus coreanae)

김승창,이승환,채한화,김의형,정기용,장선식,최봉환 충남대학교 농업과학연구소 2019 Korean Journal of Agricultural Science Vol.46 No.3

Due to the ban on the use of antibiotics, interest has been increasing for the development of therapeutic agents to treat various diseases using natural resources. Osthole, a natural coumarin compound used in traditional Chinese medicines, exerts an anti-inflammatory effect, but its effects in cows remain unknown. In this study, the effect of osthole on lipopolysaccharide (LPS)- or concanavalin-A (Con-A)- stimulated peripheral blood mononuclear cells (PBMCs) was assessed. Jugular venous blood was collected from Korean calves, and PBMCs were isolated. They were then used to study the immune response of PBMCs to treatment with osthole and LPS or Con-A for 72 h by measuring inflammatory cytokines including tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). Osthole significantly inhibited the mRNA secretion of TNF-α and IFN-γ in a dose-dependent manner. Therefore, osthole inhibited LPS- or Con-A- induced TNF-α and Con-A-induced IFN-γ production significantly in dose-dependent manner. These results clearly suggest that osthole inhibited the LPS- or Con-A- stimulated upregulation of pro-inflammatory cytokines in a dose-dependent manner, without causing obvious cytotoxic effects. Osthole could also protect cows from LPS- or Con-A- induced endotoxin shock, possibly by inhibiting the production of pro-inflammatory cytokines, which suggests that osthole might be a novel therapeutic agent for the prevention of inflammatory diseases.

Osthole Attenuates Inflammatory Responses and Regulates the Expression of Inflammatory Mediators in HepG2 Cells Grown in Differentiated Medium from 3T3-L1 Preadipocytes

Shu-Ju Wu 한국식품영양과학회 2015 Journal of medicinal food Vol.18 No.9

This study explored the anti-inflammatory mechanisms by which osthole acted on HepG2 cells cultured in a differentiated medium from cultured 3T3-L1 preadipocyte cells. HepG2 cells, a human liver cell line, were treated with various concentrations of osthole in differentiated media from cultured 3T3-L1 cells to evaluate proinflammatory cytokines, inflammatory mediators, and signaling pathways. We used enzyme-linked immunosorbent assay kits to determine the levels of proinflammatory cytokines, real-time polymerase chain reaction to assay the mRNA expression, and western blot to determine the expression of cyclooxygenase-2 (COX-2) and heme oxygenase-1 (HO-1) proteins. We also investigated inflammatory mechanism pathway members, including mitogen-activated protein kinase (MAPK) and nuclear transcription factor kappa-B (NF-κB). Osthole was able to suppress the levels of proinflammatory cytokines interleukin (IL)-1β and IL-6, as well as chemokines monocyte chemoattractant protein-1 and IL-8. In addition, COX-2 was suppressed and HO-1 expression was increased in a concentration-dependent manner. Osthole was also able to decrease IκB-α phosphorylation and suppress the phosphorylation of MAPKs. These results suggest that osthole has anti-inflammatory effects as demonstrated by the decreased proinflammatory cytokine and mediator production through suppression of the NF-κB and MAPK signaling pathways in HepG2 cells when they are incubated on the differentiated medium from 3T3-L1 cells.

The Effect of the Major Components of Fructus Cnidii on Osteoblasts In Vitro

Wenping Zhang,Dongming Ma,Qiduo Zhao,Torao Ishida 사단법인약침학회 2010 Journal of Acupuncture & Meridian Studies Vol.3 No.1

In traditional Chinese medicine, the cause of weak bones or bone loss is generally regarded as a result of kidney deficiency. Fructus Cnidii (FC), which is also known as She-Chuang-Zi, is a traditional herb that has been claimed to have kidney warming effects that invigorate Yang. In this study, we tried to determine the bone production-inducing effect of FC on osteoblastic cells in vitro using osthole, the main component of FC. Osteoblasts were isolated from neonatal Sprague-Dawley rat calvaria using the tissue piece culture method and treated with various concentrations of osthole ranging from 2.5 to 640 μg/mL, together with a blank control. Cell proliferation,alkaline phosphatase (ALP) activity, and bone nodules were measured. The cells were examined by hematoxylin-eosin staining, the Gomori Calcium-Cobalt method and immunofluorescent staining. The 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (or MTT) assay, ALP assay, and bone nodule results indicated significantly enhanced osteoblastic proliferation and differentiation at concentrations of osthole ranging from 40 to 320 μg/mL. Concentrations lower than 40 μg/mL seemed less effective, and cytotoxicity to osteoblasts was observed at concentrations higher than 320 μg/mL. These results indicate that osthole is effective at inducing osteoblastic bone formation through the up-regulation of ALP activity. FC is a Chinese herb used to treat lumbar pain in clinical practice. Further studies concerning the effects and mechanism of osthole on osteoporotic patients and animals should be performed, as these studies may lead to the development of a drug treatment for osteoporosis in the future.

Larvicidal activity of <i>Cnidium monnieri</i> fruit coumarins and structurally related compounds against insecticide‐susceptible and insecticide‐resistant <i>Culex pipiens pallens</i> and <i>Aedes aegypti</i>

Wang, Zhangqian,Kim, Jun‐,Ran,Wang, Mo,Shu, Shaohua,Ahn, Young‐,Joon John Wiley Sons, Ltd. 2012 Pest Management Science Vol.68 No.7

<P><B>Abstract</B></P><P><B>BACKGROUND:</B> An assessment was made of the toxicity of imperatorin and osthole identified in <I>Cnidium monnieri</I> fruit, 11 related compounds and five insecticides to larvae from insecticide‐susceptible <I>Culex pipiens pallens</I> (KS‐CP strain) and <I>Aedes aegypti</I> and wild <I>C</I>.<I>p</I>. <I>pallens</I> (YS‐CP colony) using a direct‐contact mortality bioassay. Results were compared with those of the conventional larvicide temephos.</P><P><B>RESULTS:</B> Imperatorin (LC<SUB>50</SUB> = 3.14 and 2.88 mg L<SUP>−1</SUP>) was 1.9‐, 3.7‐ and 4.2‐fold and 2.4‐, 4.5‐ and 4.6‐fold more toxic than isopimpinellin, isoimperatorin and osthole against susceptible <I>C. p. pallens</I> and <I>A. aegypti</I> larvae respectively. Overall, all of the compounds were less toxic than temephos (0.011 and 0.019 mg L<SUP>−1</SUP>). The toxicity of these compounds was virtually identical against larvae from the two <I>Culex</I> strains, even though YS‐CP larvae were resistant to fenthion (resistance ratio RR = 390), deltamethrin (RR = 164), cyfluthrin (RR = 14) and temephos (RR = 14). This finding indicates that the coumarins and the insecticides do not share a common mode of action. The structure–activity relationship indicates that the chemical structure and alkoxy substitution and length of the alkoxyl side chain at the C8 position are essential for imparting toxicity.</P><P><B>CONCLUSION:</B> The <I>C. monnieri</I> fruit‐derived coumarins and the related coumarins described merit further study as potential insecticides or lead molecules for the control of insecticide‐resistant mosquito populations. Copyright © 2012 Society of Chemical Industry</P>

Alcohol, NAFLD, Other : Osthol Attenuated Liver Steatosis via Decrease Liver De Novo Triglyceride Synthesis not by Insulin Resistance: Multifaceted Effects of Cindium Monnieri Extract

( Ho Hyun Nam ),( Dae Won Jun ),( Hye Joon Jeon ),( Jai Sun Lee ),( Waqar Khalid Saeed ),( Eun Kyung Kim ) 대한간학회 2013 춘·추계 학술대회 (KASL) Vol.2013 No.1

Background: Osthol is a coumarin compound isolated from the fruit of Cnidium monnieri. Previous studies showed osthol have anti-inflammatory effects on various diseases. Nowadays nonalcoholic fatty liver disease (NAFLD) has been believed as a consequence of a ‘multi-hit’ process. However, there is no multi-faceted and comprehensive evaluation about effects of osthol. The current study evaluated effects of osthol on intrahepatic fat synthesis, β-oxidation, inflammation, and insulin resistance by multifaceted-analysis. Methods: SD rats (n=30) were divided into control, NFLD, and osthol groups. NAFLD and osthol groups were fed high-fat diet for 14 weeks. After 8 weeks osthol group was supplemented orally with osthol 20mg/kg. Oral glucose tolerance test (OGTT) was performed. Immunohistochemical (4-HNE, F4/80) and H&E staining were performed on all tissue samples. SREBP1c, FAS, and SCD-1 mRNA expression were measured to assess intrahepatic fat de novo synthesis. PPAR-α, CROT, MCP-1, IRS-1, and IRS-2 expressions were assessed with RT- PCR. Results: H&E staining revealed that compared to NASH, osthol group showed significantly decreased intrahepatic fat (39.4% vs. 21.0%, P=0.021). SREBP1c expression of NAFLD group increased compared to control group (P=0.0001), while osthol treatment decreased SREBP1c expression (P=0.0059). In osthol group, intrahepatic FAS and SCD-1 expression which are down stream of SREBP1c decreased significantly compare to NAFLD Group. PPAR-α expression of osthol was also significantly higher than NAFLD (P=0.0147). However, there was no statistically difference in expression of carnitine octanoyltransferase which is marker of β-oxidation. IRS-1 and IRS-2 expression which are involving in insulin signal pathway and AUROC of OGGT were not different in NAFLD and osthol group. There was no significant difference in periportal inflammation, intrahepatic fibrosis, and kupffer cell number between the groups. Conclusions: Osthol treatment attenuated liver steatosis via decreasing liver de novo triglyceride synthesis and had nominal effects on insulin resistance and liver in- flammations.

Polymeric micelles including osthol-based drug combination for lung cancer therapy

Jae Min Lee,Yu Been Shin,Hee Ji Shin,Dae Hwan Shin 한국실험동물학회 2022 한국실험동물학회 학술발표대회 논문집 Vol.2022 No.7

Angelica속 생약의 정유성분에 관한 연구(IV) -강활의 정유성분-

지형준,김현수,Chi, Hyung-Joon,Kim, Hyun-Soo 한국생약학회 1993 생약학회지 Vol.24 No.2

Essential oil of the root of Angelica koreana Max. (Umbelliferae) was investigated. Essential oil was obtained from the dried roots by steam distillation and fractionated by column chromatography. Each isolate or fraction was identified by GC, GC-MS and spectral analysis. It was found to contain nine monoterpenes such as ${\alpha}-pinene$(7.0%), camphene, ${\beta}-pinene$, myrcene, ${\alpha}-phellandrene$, ${\Delta}-3-carene$, p-cymene, limonene(2.8%), terpinolene and also found to contain m-cresol(11.6%), citronellol, citral b, methylcinnamate, eudesmol and osthol. 2-Hydroxy-5-methylacetophenone and twelve compounds were tentatively identified.

Acute toxicity assessment of Osthol content in bio-pesticides using two aquatic organisms

Eun-Chae Yim,Hyeon Joe Kim,Seong-Jun Kim 환경독성보건학회 2014 환경독성보건학회지 Vol.29 No.-

Objectives : This study focused on the assessment of acute toxicity caused by Osthol, a major component of environment-friendly biological pesticides, by using two aquatic organisms. Methods : The assessment of acute toxicity caused by Osthol was conducted in Daphnia magna and by examining the morphological abnormalities in Danio rerio embryos. Results : The median effective concentration value of Osthol in D. magna 48 hours after inoculation was 19.3 μM. The median lethal concentration of D. rerio embryo at 96 hours was 30.6 μM. No observed effect concentration and predicted no effect concentration values of Osthol in D. magna and D. rerio were calculated as 5.4 and 0.19 μM, respectively. There was an increase in the morphological abnormalities in D. rerio embryo due to Osthol over time. Coagulation, delayed hatching, yolk sac edema, pericardial edema, and pigmentation were observed in embryos at 24–48 hours. Symptoms of scoliosis and head edema occurred after 72 hours. In addition, bent tails, ocular defects, and symptoms of collapse were observed in fertilized embryo tissue within 96 hours. Ocular defects and pigmentation were the additional symptoms observed in this study. Conclusions : Because Osthol showed considerable toxicity levels continuous toxicity evaluation in agro-ecosystems is necessary when bio-pesticides containing Osthol are used.

Acute toxicity assessment of Osthol content in bio-pesticides using two aquatic organisms

임은채,김현조,김성준 환경독성보건학회 2014 환경독성보건학회지 Vol.29 No.-

Objectives This study focused on the assessment of acute toxicity caused by Osthol, amajor component of environment-friendly biological pesticides, by using two aquatic organisms. Methods The assessment of acute toxicity caused by Osthol was conducted in Daphniamagna and by examining the morphological abnormalities in Danio rerio embryos. Results The median effective concentration value of Osthol in D. magna 48 hours afterinoculation was 19.3 μM. The median lethal concentration of D. rerio embryo at 96hours was 30.6 μM. No observed effect concentration and predicted no effect concentrationvalues of Osthol in D. magna and D. rerio were calculated as 5.4 and 0.19 μM,respectively. There was an increase in the morphological abnormalities in D. rerio embryodue to Osthol over time. Coagulation, delayed hatching, yolk sac edema, pericardialedema, and pigmentation were observed in embryos at 24–48 hours. Symptoms of scoliosisand head edema occurred after 72 hours. In addition, bent tails, ocular defects,and symptoms of collapse were observed in fertilized embryo tissue within 96 hours. Ocular defects and pigmentation were the additional symptoms observed in this study. Conclusions Because Osthol showed considerable toxicity levels continuous toxicity evaluationin agro-ecosystems is necessary when bio-pesticides containing Osthol are used.