Fructus Cnidii Research Articles

Effects of osthole microemulsion by nasal administration on the cholinergic pathway in mice treated with scopolamine

ObjectivesOsthole is the main active component of Fructus Cnidii, which is the dry ripe fruit of Cnidium monnieri (L.) Cuss. and has long been used in clinic practice with pharmacological activity in the central nervous system (CNS). However, Osthole exhibits low bioavailability, fast distribution and elimination, and low concentrations in the brain when given orally. In this study, we aimed to develop a new dosage form to increase the concentrations of osthole in the brain and enhance its pharmacological effects in the CNS through reducing the dosage and improving the stability and bioavailability. Thus, osthole microemulsion was prepared to investigate its effects in Alzheimer’s disease model mice.MethodsOsthole microemulsion was prepared, and the particle size and distribution were detected. The effects of osthole microemulsion by nasal administration on learning and memory abilities in scopolamine‐treated mice were assessed by behavioural tests. The superoxide dismutase (SOD) activity, glutathione (GSH) level and malondialdehyde (MDA) content in the serum were examined. Choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) expressions in the olfactory‐basal forebrain were detected. The acetylcholine (ACh) level and the histological morphology in the brain were also measured.ResultsThe average particle size of 1μg/μl osthole microemulsion was less than 15 nm. It was characterized as sphere under the transmission electron microscopy, and the osthole was completely encapsulated in the microemulsion core. Morris water maze and novel object recognition tests showed that osthole microemulsion improved spatial and object learning and memory in scopolamine‐treated mice. Moreover, osthole microemulsion restored the abnormal activity of SOD and increased the levels of MDA and GSH in the serum. Brain immunohistochemistry staining showed that osthole microemulsion up‐regulated ChAT expression, while down‐regulated AChE in the olfactory‐basal forebrain cholinergic pathway. Additionally, the ACh level and pathological morphology in the brain were also reversed after nasal administration with osthole microemulsion.ConclusionThe concentration of 1μg/μl of osthole microemulsion is an ideal dosage form with small particle size, uniform distribution, and high permeation. Osthole microemulsion ameliorates memory impairment induced by scopolamine in mice, likely via the olfactory‐basal forebrain cholinergic pathway and by ameliorating oxidative stress. This may have an implication for the development of intranasal brain targeting drugs via the nose‐brain pathway as potential treatments of certain CNS disorders.Support or Funding InformationThe present study was supported by research grants from the National Natural Science Foundation of China (grant no. 81773717 and 81703901), the Shandong Province Natural Science Foundation of China (grant no. ZR2016HB56), the Foundation of Overseas Distinguished Taishan Scholars of Shandong Province, and he Taian Municipal Science and Technology Bureau funding (grant no: 2016NS1078).

Osthole-loaded N-octyl-O-sulfonyl chitosan micelles (NSC-OST) inhibits RANKL-induced osteoclastogenesis and prevents ovariectomy-induced bone loss in rats.

Osthole (OST), a derivative of Fructus Cnidii, has been proved to have potential anti‐osteoporosis effects in our recent studies. However, its pharmacological effects are limited in the human body because of poor solubility and bioavailability. Under the guidance of the classical theory of Chinese medicine, Osthole‐loaded N‐octyl‐O‐sulfonyl chitosan micelles (NSC‐OST), which has not previously been reported in the literature, was synthesized in order to overcome the defects and obtain better efficacy. In this study, we found that NSC‐OST inhibited on the formation and resorption activity of osteoclasts through using a bone marrow macrophage (BMM)‐derived osteoclast culture system in vitro, rather than affecting the viability of cells. We also found that NSC‐OST inhibited osteoclast formation, hydroxyapatite resorption and RANKL‐induced osteoclast marker protein expression. In terms of mechanism, NSC‐OST suppressed the NFATc1 transcriptional activity and the activation of NF‐κB signalling pathway. In vivo, ovariectomized (OVX) rat models were established for further research. We found that NSC‐OST can attenuate bone loss in OVX rats through inhibiting osteoclastogenesis. Consistent with our hypothesis, NSC‐OST is more effective than OST in parts of the results. Taken together, our findings suggest that NSC‐OST can suppress RANKL‐induced osteoclastogenesis and prevents ovariectomy‐induced bone loss in rats and could be considered a safe and more effective anti‐osteoporosis drug than OST.

Antipruritic Effect of Ethyl Acetate Extract from Fructus cnidii in Mice with 2,4-Dinitrofluorobenzene-Induced Atopic Dermatitis.

Atopic dermatitis (AD) is a common inflammatory skin disease characterized by intense pruritus and skin lesions. The exact cause of AD is not yet known and the available therapeutic strategies for AD are limited. Fructus cnidii is commonly used in traditional Chinese medicine as an herb for treating chronic itch. However, the mechanism underlying the antipruritic effects of Fructus cnidii is not well understood. In the present study, we investigated the antipruritic effect of locally administered ethyl acetate extract from Fructus cnidii (EAEFC) to 2,4-dinitrofluorobenzene- (DNFB-) induced AD in a mouse model. The scratching behavior, skin thickness, dermatitis score, weight, blood immunoglobulin E (IgE) level, and itch-related cytokine levels were subsequently monitored and evaluated. Results showed that EAEFC treatment attenuated the DNFB-induced AD-like symptoms by alleviating the skin lesions and decreasing the dermatitis score. Hematoxylin and eosin (H&E) and toluidine blue (TB) staining analyses demonstrated that EAEFC mitigated the DNFB-induced increase in skin thickness and prevented the infiltration of mast cells. Behavioral tests showed that EAEFC decreased the DNFB-induced acute and chronic scratching behaviors. Furthermore, EAEFC reduced the levels of itch-related cytokines, such as thymic stromal lymphopoietin (TSLP), interleukin- (IL-) 17, IL-33, and IL-31, and the DNFB-induced boost in serum IgE. Collectively, these results suggest that EAEFC is a potential therapeutic candidate for the treatment of chronic itch in AD.

Osthole ameliorates cartilage degradation by downregulation of NF-κB and HIF-2α pathways in an osteoarthritis murine model

Osteoarthritis (OA) is a common and disabling joint disease mainly characterized by cartilage degradation, with the knees most commonly affected. No effective treatment for the cartilage degradation of OA exists. Preliminary studies have revealed the protective and osteogenic effects of osthole, a natural coumarin first isolated from Cnidium monnieri (Fructus Cnidii); however, no evidence of osthole in an OA-related model has been published to date. This study further explored the effects of osthole in a monoiodoacetate (MIA)-induced OA-related animal model and focused on the molecular mechanism(s) behind the anti-inflammatory and cartilage protective effects of osthole. This study revealed that the cartilage protective effect of osthole in a MIA-induced osteoarthritis (OA) murine model can be explained by downregulation of COX-2 and RUNX2 by inhibition of NF-κB and HIF-2α up-regulated by OA induction, resulting in downregulation of MMP-13, Syndecan IV and ADAMTS-5. In addition, osthole might have anti-inflammatory and analgesic effects due to COX-2 inhibition. Osthole can be considered as a potential component of the treatment of OA, for it possesses a cartilage protective effect, as well as anti-inflammation, analgesic, and movement improving effects. Further preclinical and human clinical studies are needed to examine the efficacy and safety profile of long-term therapy.

Osthole inhibits osteoclasts formation and bone resorption by regulating NF-κB signaling and NFATc1 activations stimulated by RANKL.

Chinese herbal medicine Fructus Cnidii has an outstanding effect on chronic lumbar pain and impotence, also has been used against osteoporosis with high frequency. Yet, the mechanisms of osthole, a derivative of Fructus Cnidii, on osteoclasts remains barely known. In this study, it was found out that osthole (10-6 mol/L, 10-5 mol/L) had the influence of inhibiting osteoclast formation and bone resorptive activities induced by receptor activator of nuclear factor κB ligand (RANKL), rather than affecting the viability of osteoclast-like cells. Furthermore, osthole could also inhibit the messenger RNA expressions of c-Src, tartrate-resistant acid phosphatase, β3-Integrin, matrix metallopeptidase 9, and cathepsin K. The results of the mechanistic study indicated that osthole regulated the nuclear factor of activated T-cells cytoplasmic 1 (NFATc1) and nuclear factor-κB (NF-κB) activations following the RANKL stimulation. These findings suggested that the inhibitory effects of osthole were associated with restraining the activations of NFATc1 and NF-κB induced by RANKL. Thus osthole can be used as a potential treatment for abnormal bone-resorption related diseases.

Effects of pungent essential oil from three Chinese herbs on percutaneous absorption of alkaloids from Sophorae Flavescentis Radix

To investigate the effects of essential oil from three kinds of pungent herbs,namely Menthae Haplocalycis Herba,Atractylodis Rhizoma and Cnidii Fructus,on the transdermal absorption in vitro of alkaloids from Sophorae Flavescentis Radix. The modified vertical Franz diffusion cell was used to conduct a transdermal experiment in vitro with the isolated abdominal skin of the SD rats as the transdermal absorption barrier. The effects of such three kinds of pungent essential oil on percutaneous absorption of alkaloids from Sophorae Flavescentis Radix were investigated by determining the content of 6 alkaloids( oxymatrine,oxysophocarpine,N-methylcytisine,sophoridine,matrine,and sophocarpidine) in the transdermal acceptor with ultra-performance liquid chromatography-triple quadruple mass spectrometry( UPLC-TQ-MS) technique simultaneously. With enhancement ratio( ER) as the index,their effects on promoting penetration was as follows: 1% Atractylodis Rhizoma oil > 1% Cnidii Fructus oil > 3% Azone ≈ 3% Atractylodis Rhizoma oil > 5%Atractylodis Rhizoma oil > 3% Cnidii Fructus oil ≈ 5% Cnidii Fructus oil > 3% Menthae Haplocalycis Herba oil > 5% Menthae Haplocalycis Herba oil > 1% Menthae Haplocalycis Herba oil > Blank. The results showed that these three kinds of pungent essential oil could be used as enhancers for alkaloids of Sophorae Flavescentis Radix,providing scientific guidance for improving percutaneous absorption of alkaloids from Sophorae Flavescentis Radix.

Osthole prevents tamoxifen-induced liver injury in mice

Tamoxifen (TMX) is an antiestrogen drug that is used in the treatment and prevention of all stages of estrogen-dependent breast cancer. Adverse effects of TMX include hepatotoxicity. In this study, we investigated the therapeutic effects of osthole, isolated from medicinal plants especially Fructus Cnidii, on TMX-induced acute liver injury in mice. Mice were injected with osthole (100 mg/kg, ip) or vehicle, followed by TMX (90 mg/kg, ip) 24 h later. We showed that a single injection of TMX-induced liver injury and oxidative stress. Pretreatment with osthole attenuated TMX-induced liver injury evidenced by dose-dependent reduction of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities. Pretreatment with osthole also blunted TMX-induced oxidative stress, evidenced by significant increase of reduced glutathione (GSH) as well as reduction of malondialdehyde (MDA) and hydrogen peroxide (H2O2). Consistently, osthole significantly enhanced the expressions of antioxidant genes (GPX1, SOD2, GCL-c, and G6pdh), but suppressed those of pro-oxidant genes (NOX2 and ACOX). Furthermore, osthole inhibited the production of inflammatory cytokines, reduced the metabolic activation of TMX, and promoted its clearance. We further revealed that osthole elevated hepatic cAMP and cGMP levels, but inhibition of PKA or PKG failed to abolish the hepatoprotective effect of osthole. Meanwhile, prominent phosphorylation of p38 was observed in liver in response to TMX, which was significantly inhibited by osthole. Pretreatment with SB203580, a p38 inhibitor, significantly attenuated TMX-induced increase of ALT and AST activities, reduced oxidative stress, and reversed the alterations of gene expression caused by TMX. Moreover, pretreatment with L-buthionine sulfoximine (BSO), an inhibitor of GSH synthesis, partly reversed the effect of osthole on TMX-induced liver injury. Consistently, pretreatment with N-acetyl-L-cysteine (NAC) significantly attenuated TMX-induced increase in ALT and AST activities. Notably, both BSO and NAC had no detectable effect on the phosphorylation levels of p38. Collectively, our results suggest that osthole prevents TMX hepatotoxicity by suppressing p38 activation and subsequently reducing TMX-induced oxidative damage.

Qualitative and quantitative study of Shengui anti-pruritic mixture

Objective To study the qualitative and quantitative standard of Shengui anti-pruritic mixture. Mehtods Sophorae Flavescentis Radix, Cnidii Fructus, Polygoni Multiflori Radix, Astragali Radix, Paeoniae Radix Alba and Glycyrrhizae Radix Et Rhizoma were identified by TLC. The contents of matrine and ferulic acid were determined by HPLC. Results The TLC showed the obvious characteristics with the spots clear and well-separated with no interference by negative control. Matrine showed a good linear relationship at 0.062 5-2.500 0 μg (r=0.999 9) with a good meeting of adding sample recovery test. Ferulic acid showed a good linear relationship at 0.021 0-0.840 0 μg (r=0.999 9) with a good meeting of adding sample recovery test. Conclusions The method is easy-operated and accurate which showed a good specificity for the quality control of Shengui anti-pruritic mixture. Key words: Chromatography, thin layer; Chromatography, high pressure liquid; Quality control; Matrine; Ferulic acid; Shengui anti-pruritic mixture

Review for treatment effect and signaling pathway regulation of kidney-tonifying traditional Chinese medicine on osteoporosis

The treatment effect and signaling pathway regulation effects of kidney-tonifying traditional Chinese medicine on osteoporosis have been widely studied, but there is no systematic summary currently. This review comprehensively collected and analyzed the traditional Chinese medicines on the treatment and signaling pathway regulation of osteoporosis in recent ten years, such as Epimedii Folium, Drynariae Rhizoma, Cnidii Fructus, Eucommiae Cortex, Psoraleae Fructus and Dipsaci Radix. Based on the existing findings, the following conclusions were obtained: ①kidney-tonifying traditional Chinese medicine treated osteoporosis mainly through BMP-Smads, Wnt/β-catenin, MAPK, PI3K/AKT signaling pathway to promote osteoblast bone formation and through OPG/RANKL/ RANK, estrogen, CTSK signaling pathway to inhibit osteoclasts of bone resorption. Epimedii Folium, Drynariae Rhizoma, Cnidii Fructus and Psoraleae Fructus up-regulated the expression of key proteins and genes of BMP-Smads and Wnt/β-catenin signaling pathways to promote bone formation. Epimedii Folium, Drynariae Rhizoma, Cnidii Fructus, Eucommiae Cortex, Psoraleae Fructus and Dipsaci Radix inhibited the bone resorption by mediating the OPG/RANKL/RANK signaling pathway. ②Kidney-tonifying traditional Chinese medicine prevented and treated osteoporosis through a variety of ways: icariin in Epimedii Folium, naringin in Drynariae Rhizoma, osthole in Cnidii Fructus and psoralen in Psoraleae Fructus can regulate BMP-Smads, Wnt/β-catenin signaling pathway to promote bone formation, but also activate OPG/RANKL/RANK, CTSK and other signaling pathways to inhibit bone resorption. ③The crosstalk of the signaling pathways and the animal experiments of the traditional Chinese medicine on the prevention and treatment of osteoporosis as well as their multi-target mechanism and comprehensive regulation need further clarification.

Analgesic and Anti-Inflammatory Effects of Herbal Mixture Extracts in Mice

Herbal mixture extracts (HME) comprised of Semen Sojae Nigrum, Fructus Cnidii, Radix Glycyrrhizae, and Cornu Cervi. Herein, we employed three mouse models, including hot-plate test, acetic acid (AA)-induced writhing test and AA-induced vascular permeability test, to determine analgesic and anti-inflammatory effects of HME. Results revealed that HME exhibited analgesic effects in hot-plate test and in AA-induced writhing test, as evidenced by increasing the latency to lick paws and decreasing AA-induced writhing counts, respectively. HME also significantly and dose-dependently decreased AA-induced vascular permeability, indicating HME exhibited anti-inflammatory effects. Similar improvement can be observed in aspirin treatment that used as positive control in this study. Most of medicinal effects of Fructus Cnidii are considered to attribute to coumarins, such as osthol (OST) and imperatorin (IMP) with several pharmacological activities. We then used OST and IMP as bioactive components in HME. The content of OST and IMP in HME was 3.57 ± 0.10 mg/g and 1.20 ± 0.02 mg/g, respectively, from three independent batches. Only OST possessed inhibitory effects in three mouse models, suggesting that OST may partially involve in protective effects of HME. These results demonstrated that HME has a potential on anti-analgesic effects and anti-inflammatory actions.

Osthole promote differentiation and inhibit proliferation of osteoblast by activating wnt signaling and endoplasmic reticulum stress

Background: Osthole is extracted from Fructus Cnidii and is proved to be effective in the treatment of osteoporosis in rats. However, data are still scarce and the mechanism remains elusive. Objective: To investigate the effect of Osthole on proliferation and differentiation of osteoblast. Materials and Methods: Cells were divided into five groups: control group, β-estradiol group (10−8 M), and Osthole groups (10−6 M, 10−5 M, and 10−4 M). Osteoblast proliferation was evaluated by Cell Counting Kit-8 (CCK-8) assay. Alkaline phosphatase (ALP) activity was detected by ALP staining and enzymatic measurement. Mineralization was detected by alizarin-red staining. The level of osteocalcin was measured by enzyme-linked immunosorbent assay (ELISA). The expression of key proteins of Wnt/β-catenin signaling pathway and endoplasmic reticulum stress (ERS) was analyzed by Western blot. Results: Cell proliferation was retarded in moderate-dose and high-dose Osthole group at 1d, 2d and 3d and in low-dose Osthole group at 1d and 2d (P Abbreviations used: ALP: Alkaline phosphatase; ELISA: Enzyme-linked immunosorbent assay; CCK-8: Cell Counting Kit-8; ERS: endoplasmic reticulum stress; DMEM: Dulbecco's Modified Eagle Medium; α-MEM: α-minimum essential medium; EDTA: trypsin/ethylenediaminetetraacetic acid; DMF: N,N-dimethylformamide; TBS-T: Tris-buffered saline-Tween 20; RIPA: Radio Immuno Precipitation Assay; HRP: Horseradish peroxidase; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; BCA: Bicinchoninic acid; ANOVA: Analysis of variance.

Antifungal Activity of Osthole on Microsporum canis through Interfering with Biosynthesis of Fungal Cell Wall

Osthole extracted from Cnidii Fructus, a herb used in traditional Chinese medicine, was reported to possess multiple activities such as antibacterial, antiinflammatory, antitumor and insecticidal. However, its antifungal activity against Microsproum canis has not been investigated yet. The present study aimed to investigate the antifungal effect of osthole against Microsproum canis. The minimum inhibitory concentration and a growth inhibitory curve of osthole on Microsporum canis were according to the program of M38-A2 of Clinical and Laboratory Standards Institute. HPLC analysis, aniline blue test and spectrophotometry were used to determine contents of ergosterol, 1,3-β-D-glucan and chitin from Microsporum canis, respectively. The results indicated that minimum inhibitory concentration of osthole against Microsporum canis was 1.95 µg/ml. HPLC analysis demonstrated that there was no significant change in the content of ergosterol between the osthole and control group. Compared with the control group, the content of 1,3-β-D-glucan and chitin in the cell wall of Microsporum canis was decreased. Taken together, this research showed that osthole could notably inhibit the growth of Microsporum canis and the antifungal activity was related to inhibiting the biosynthesis of 1,3-β-D-glucan and chitin in the cell wall of Microsporum canis, making osthole a potential future antifungal agent.

Synergic Anti-Pruritus Mechanisms of Action for the Radix Sophorae Flavescentis and Fructus Cnidii Herbal Pair.

Radix Sophorae Flavescentis (RSF) and Fructus Cnidii (FC) compose a typical herbal synergic pair in traditional Chinese medicine (TCM) for pruritus symptom treatments. The mechanisms of action for the synergy are not understood. This paper aims at predicting the anti-pruritus targets and the main active ingredients for the RSF and FC herbal pair. We demonstrate that the RSF–FC herbal pair can be elucidated by mining the chemical structures of compounds derived from RSF and FC. Based on chemical structure data, the putative targets for RSF and FC were predicted. Additional putative targets that interact with the anti-pruritus targets were derived by mapping the putative targets onto a PPI network. By examining the annotations of these proteins, we conclude that (1) RSF’s active compounds are mainly alkaloids and flavonoids. The representative putative targets of the alkaloids are inflammation-related proteins (MAPK14, PTGS2, PTGS2, and F2) and pruritus-related proteins (HRH1, TRPA1, HTR3A, and HTR6). The representative putative targets of the flavonoids are inflammation-related proteins (TNF, NF-κB, F2, PTGS2, and PTGS2) and pruritus-related proteins (NR3C1 and IL2). (2) FC’s active compounds are mainly coumarins. Their representative putative targets are CNS-related proteins (AChE and OPRK1) and inflammation-related proteins (PDE4D, TLR9, and NF-κB). (3) Both RSF and FC display anti-inflammatory effects, though they exhibit their anti-pruritus effects in different ways. Their synergy shows that RSF regulates inflammation-related pruritus and FC regulates CNS-related pruritus.

A fast and accurate method for the pharmacokinetic research of four coumarin analogs in Fructus cnidii using capillary electro-chromatography with a methacrylate ester-based monolithic column.

In the present study, a monolithic capillary column with higher permeability was developed for the in vivo discrimination of four coumarin analogs (bergapten, 2'-acetylangelicin, imperatorin, and osthole) that typically require long separation times in HPLC. Instead of conventional methacrylate ester monolith (containing 19.5% porogen) with insufficient permeability (K=1.52 - 1.66×10-14 M2 ) for plasma sample, the proposed column (20.5% porogen) had better permeability (around 3.80×10-14 M2 ) while properties such as pore distribution, stability, and resolution changed slightly. As a result, due to the negatively charged electro-dynamic flow of the methacrylate ester groups in the monolith, the migration of targeted analytes was achieved within 6min (compared with 30min in HPLC) with acceptable resolution and improved sensitivity (0.005-0.02μg/mL vs. 0.04μg/mL). The proposed method was also applied to pharmacokinetic research: accelerated solvent extraction (ASE) was used to improve the extraction efficiency, which prepared extract much faster and more pure than conventional methods. As the pharmacokinetic parameters indicated, the monolithic capillary electro-chromatography method was efficient, sensitive, specific, and durable, guaranteeing its utility for the determination of multiple structure-related compounds in rat plasma.

Isolation and purification of osthole and imperatorin from Fructus Cnidii by semi-preparative supercritical fluid chromatography

ABSTRACTFructus Cnidii, the dried ripe fruit of Cnidium monnieri (L.) Cusson., has been widely used in traditional Chinese medicine. Osthole and imperatorin are the main active ingredients of Fructus Cnidii and had been found of antispasmodic, anti-HIV, anti-fungal, anti-viral, anti-tumor, anti-mutagenic, anti-arrhythmic, hypotensive, and broad-spectrum antimicrobial effects. A supercritical fluid chromatography (SFC) method for isolation and purification of osthole and imperatorin from Fructus Cnidii was established in this work. The separation conditions, including the stationary phase, the organic modifier, the composition and the flow rate of the mobile phase, column backpressure and column temperature, were optimized on analytical scale at first. And then a semi-preparative SFC (SP-SFC) method was developed based on the conditions of analytical scale SFC. SP-SFC was accomplished on YMC-Pack NH2 column. Ethanol was used as the modifier and its percentage in the mobile phase was 3%. The flow rate of the mobile phase was 20 mL/min, column backpressure was 13 MPa, column temperature was 318 K, detection wavelength was 310 nm, and injection volume was 0.2 mL. Under the optimum conditions, osthole and imperatorin were obtained with high purities as determined by high performance liquid chromatography. The chemical structures of the obtained compounds were identified by nuclear magnetic resonance and mass spectrum analysis.

Osthole Protects Bone Marrow-Derived Neural Stem Cells from Oxidative Damage through PI3K/Akt-1 Pathway

In recent years, neural stem cell (NSC) transplantation has been widely explored as a treatment for neurodegenerative diseases. NSCs are special cells that have some capacity for self-renewal and the potential to differentiate into multiple cell types. However, the inflammatory environment of diseased tissue is not conducive to the survival of transplanted cells. Osthole (Ost) is a principal bioactive component of Fructus Cnidii, Radix Angelicae Pubescentis and other traditional Chinese medicines. Ost has a wide range of pharmacological activities, such as anti-inflammation, immunomodulation, and neuroprotection. In the present study, we assessed the protective effects of Ost on bone marrow-derived-NSCs (BM-NSCs) against injury induced by hydrogen peroxide (H2O2). BM-NSCs were pre-treated with different doses of Ost and treated with H2O2. The cell counting kit-8 (CCK-8) method and lactate dehydrogenase (LDH) leakage assay were used to determine cell viability. Using the TUNEL assay and RT-PCR, we evaluated the effect of Ost on cell apoptosis. The results showed that Ost had protective effects against H2O2-induced cell damage, and the number of apoptotic cells was significantly decreased in the Ost pre-treated groups compared to the H2O2 group. The expression ratio of Bax/Bcl-2 mRNA was also decreased. Furthermore, western blotting was used to analyze levels of proteins related to PI3K/Akt-1 signaling pathway, and results indicated that ost can increase p-Akt and PI3K. Our findings suggested that Ost protects BM-NSCs against oxidative stress injury, and it can be used to improve the inflammatory environment of neurodegenerative diseases so and promote the survival rate of transplanted NSCs.

Effects of Plants on Osteogenic Differentiation and Mineralization of Periodontal Ligament Cells: A Systematic Review.

This systematic review aimed to evaluate the effects of plants on osteogenic differentiation and mineralization of human periodontal ligament cells. The included studies were selected using five different electronic databases. The reference list of the included studies was crosschecked, and a partial gray literature search was undertaken using Google Scholar and ProQuest. The methodology of the selected studies was evaluated using GRADE. After a two-step selection process, eight studies were identified. Six different types of plants were reported in the selected studies, which were Morinda citrifolia, Aloe vera, Fructus cnidii, Zanthoxylum schinifolium, Centella asiatica, and Epimedium species. They included five types of isolated plant components: acemannan, osthole, hesperetin, asiaticoside, and icariin. In addition, some active substances of these components were identified as polysaccharides, coumarins, flavonoids, and triterpenes. The studies demonstrated the potential effects of plants on osteogenic differentiation, cell proliferation, mineral deposition, and gene and protein expression. Four studies showed that periodontal ligament cells induce mineral deposition after plant treatment. Although there are few studies on the subject, current evidence suggests that plants are potentially useful for the treatment of periodontal diseases. However, further investigations are required to confirm the promising effect of these plants in regenerative treatments.

Morphological and Anatomical Characteristics of Medicinal Fructus in Apiaceae

Background : As the characterization of medicinal plants is an important aspect of traditional Asian herbal medicine, this study examines the morphological and anatomical characteristics in four fructus form medicinally important plants belonging to the family Apiaceae. Methods and Results : Fruit morphology of four genera was examined using microtome sections and light microscopy. The Anethi Fructus, Anethum graveolens (Siraja) has unique wing-like and membranous lateral ribs that are, approximately wide. The Coriandri Fructus, Coriandrum sativum (Hoyuja) does not have oil ducts at the dorsal region of the mericarp and differs in the development of the dorsal ribs. The ribs appear upon drying and therefore, consist of primary and secondary ribs. The Foeniculi Fructus, Foeniculum vulgare (Hoehyang) characteristically develops three dominant dorsal ribs and has a strong aromatic fragrance. Finally, the Cnidii Fructus, Torilis japonica (Sasangja) has many oil ducts at the dorsal and carpophores regions and develops many hooked trichomes one the fructus surface. Conclusions : We conclude from this study that each plant has several prominent and distinguishing morphological and anatomical characteristics. Therefore, fruit morphology is very useful for identifying these medicinal plants. In addition, the correct use and herbal name need to be standardized for plants used traditionally in Asian medicine within Korea, China, and Japan.

The Antioxidant Activity of Cnidii Fructus and Torilis Fructus in Leydig cells

Objectives : The purpose of this study was to estimate the antioxidant activity of water extract of Cnidii Fructus (CF) and Torilis Fructus (TF) in Leydig cells. Methods : Free radical scavenging activity of CF and TF against 2,2-diphenyl-1-picrylhydrazyl (DPPH) was determined spectrophotometrically. We investigated the effect of CF and TF in Leydig cells by MTT assay. The protective effects of CF and TF against hydrogen peroxide-induced oxidative stress in Leydig cells. Superoxide dismutase (SOD), and catalase activity assays were performed in Leydig cells. Results : The results showed that CF scavenged DPPH radical in a dose-dependent manner by up to 81.2%, TF scavenged DPPH radical in a dose-dependent manner by up to 63.8%. CF showed cell viability as 121.0, 132.7, 126.6% in 5, 10, 100 μg/ml concentrations. TF showed cell viability as 127.5, 111.8% in 5, 100 μg/ml concentraions, respectively. The hydrogen peroxide-induced cytotoxicity of Leydig cells were protected to 86.3% by CF at concentration of 10 μg/ml and protected to 83.5% by TF at concentration of 100 μg/ml. Both CF and TF at all concentrations, SOD activity was not significantly changed. Catalase activity was significantly increased at 10, 100 μg/ml concentrations of CF, respectively. TF's catalase activity showed no significant difference from that of the control. Conclusions : These results suggest that CF, as an antioxidant, protects Leydig cells in hydrogen peroxide-induced oxidative stress. know that『Kwangjebikeup』played a role in settlement and spreading of foreign knowledge to civilians.

Cytochrome P450 isoenzymes in rat and human liver microsomes associate with the metabolism of total coumarins in Fructus Cnidii.

Fructus Cnidii (Cnidium) is isolated from the dry and ripe fruit of Cnidium monnier (L.) Cuss (umbelifera), an annual herb. It is demonstrated that the active constituents of Fructus Cnidii are coumarins, known as Total Coumarins of Cnidium Monnier (TCCM). Osthole (Ost) and imperatorin (Imp) are the most active constituents of TCCM which are usually regarded as the quality indicators of medicinal Fructus Cnidii. The aim is to study the metabolism of Fructus Cnidii effective monomer osthole and imperatorin in vitro by liver microsomes. CYP3A4 inhibitor ketoconazole, CYP2D6 inhibitor qunidine, CYP2C8 inhibitor trimethoprim, CYP2C9 inhibitor sulfaphenazole, and CYP1A2 inhibitor α-naphthoflavone were used to investigate the metabolism from incubation time, substrate concentration and liver microsomal concentration, respectively. The concentration of liver microsomes was 0.2mg/ml. Ost (0.8/3.2/12.8uM) was incubated at 37°C for 20min while Imp (1.6/6.4/19.2uM) was incubated for 30min. Qunidine, trimethoprim and α-naphthoflavone could significantly inhibit the disappearance of Imp; meanwhile ketoconazole, sulfaphenazole and qunidine could inhibit the disappearance of Ost. CYP1A, CYP2C are involved in the metabolism of Imp and CYP3A mediates the metabolism of Ost in rat liver microsomes. In human liver microsomes, CYP1A2, CYP2C8, CYP2D6 are involved in the metabolism of Imp; CYP3A4 is involved in the metabolism of Ost at all the tested concentrations of Ost, while CYP2C9, CYP2D6 mediate the metabolism at high concentration of Ost.