Pharmacological Effects and Molecular Mechanism of Osthole

Osthole has various pharmacological effects such as anti-cancer,anti-inflammation,prevention and treatment of cardiovascular diseases and neuroprotection.This paper reviews the advances in the research of the pharmacological effects and molecular mechanisms of osthole,in order to provide new ideas for further research and clinical application of osthole.

Experimental study of osthole on treatment of hyperlipidemic and alcoholic fatty liver in animals

AIM： To evaluate the effects of osthole on fatty liver, and investigate the possible mechanism. METHODS： A quail model with hyperlipidemic fatty liver and rat model with alcoholic fatty liver were set up by feeding high fat diet and alcohol, respectively. These experimental animals were then treated with osthole 5-20 mg/kg for 6 wk, respectively. Whereafter, the lipid in serum and hepatic tissue, and coefficient of hepatic weight were measured. RESULTS： After treatment with osthole the levels of serum total cholesterol （TC）, triglyceride （TG）, lower density lipoprotein-cholesterol （LDL-C）, coefficient of hepatic weight, and the hepatic tissue contents of TC and TG were significantly decreased. The activity of superoxide dismutase （SOD） in liver was improved. In alcohol-induced fatty liver rats, the level of malondialdehyde （MDA） in liver was decreased. In high fat-induced fatty liver quails, glutathione peroxidase （GSH-PX） in liver was significantly improved. The histological evaluation of liver specimens demonstrated that the osthole dramatically decreased lipid accumulation. CONCLUSION： These results suggested that osthole had therapeutic effects on both alcohol and high fatinduced fatty liver. The mechanism might be associated with its antioxidation.

Osthole inhibits proliferation of human breast cancer cells by inducing cell cycle arrest and apoptosis

Recent studies have revealed that osthole,an active constituent isolated from the fruit of Cnidium monnieri（L.） Cusson,a traditional Chinese medicine,possesses anticancer activity.However,its effect on breast cancer cells so far has not been elucidated clearly.In the present study,we evaluated the effects of osthole on the proliferation,cell cycle and apoptosis of human breast cancer cells MDA-MB 435.We demonstrated that osthole is effective in inhibiting the proliferation of MDA-MB 435 cells,The mitochondrion-mediated apoptotic pathway was involved in apoptosis induced by osthole,as indicated by activation of caspase-9 and caspase-3 followed by PARP degradation.The mechanism underlying its effect on the induction of G1 phase arrest was due to the up-regulation of p53 and p21 and down-regulation of Cdk2 and cyclin D1 expression.Were observed taken together,these findings suggest that the anticancer efficacy of osthole is mediated via induction of cell cycle arrest and apoptosis in human breast cancer cells and osthole may be a potential chemotherapeutic agent against human breast cancer.

Osthole improves synaptic plasticity in the hippocampus and cognitive function of Alzheimer's disease rats via regulating glutamate

Osthole, an effective monomer in Chinese medicinal herbs, can cross the blood-brain barrier and protect against brain injury, with few toxic effects. In this study, a rat model of Alzheimer＇s disease was established after intracerebroventricular injection of β-amyloid peptide （25-35）. Subsequently the rats were intraperitoneally treated with osthole （12.5 or 25.0 mg/kg） for 14 successive days. Results showed that osthole treatment significantly improved cognitive impairment and protected hippocampal neurons of AIzheimer＇s disease rats. Also, osthole treatment alleviated suppressed long-term potentiation in the hippocampus of Alzheimer＇s disease rats. In these osthole-treated Alzheimer＇s disease rats, the level of glutamate decreased, but there was no significant change in y-amino-butyric acid. These experimental findings suggest that osthole can improve learning and memory impairment, and increase synaptic plasticity in Alzheimer＇s disease rats. These effects of osthole may be because of its regulation of central glutamate and y-amino-butyric acid levels.

NP-3 Effects of Osthole Microemulsion by Nasal Administration on the Cholinergic Pathway in AD Mice

Objectives:Fructus Cnidii is the dry ripe fruit of Cnidium monnieri(L.)Cuss.,which belongs to the umbelliferous plant.It has long been used in clinic practice and has been found to have pharmacological activity in the central nervous system.Osthole is the main active component of Fructus Cnidii.However,it shows low bioavailability,fast distribution and elimination,and low concentration in the brain when given orally.In this study,we aimed to develop a new dosage form to increase the osthole concentration in the brain and enhance its pharmacological effects in the central nervous system through reducing the dosage while improving the stability and bioavailability.Thus,microemulsion containing osthole was prepared and the effects of osthole microemulsion were examined in the mouse model of Alzheimer’s disease(AD).Methods:On the basis of pseudo-ternary phase diagram,microemulsion was prepared by using polyoxyethlated Cremophor RH40 as emulsifiers,propylene glycol as assistant emulsifiers and ethyl acetate as the oil phase.The particle size and distribution of osthole microemulsion were detected by laser particle size analyzer and transmission election microscope.The content of osthole was determined by UV spectrophotometry.The effects of osthole microemulsion by nasal administration on the learning and memory abilities in scopolamine-treated mice were assessed by Morris water maze and novel object recognition tests.The superoxide dismutase(SOD)activity,glutathione(GSH)levels and malondialdehyde(MDA)contents in the serum were examined to evaluate the oxidant stress.Choline acetyltransferase(ChAT)and acetylcholinesterase(AChE)expression in the olfactory-basal forebrain pathway were detected by immunohistochemical analysis.We also investigated the acetylcholine(ACh)levels and the histological morphology in the brain.Results:The average particle size of 1μg·μL-1 osthole microemulsion was less than 15 nm.It was characterized as spheres 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 levels and pathological morphology in the brain were also reversed after nasal administration with osthole microemulsion.Conclusion:The 1μg·μL-1 osthole microemulsion is an ideal dosage form with a small particle size,uniform distribution and high permeation.Osthole microemulsion ameliorated memory impairment in scopolamine-teated mice,likely via the olfactory-basal forebrain cholinergic pathway and by reducing oxidative stress.The results implicate the development of intranasal brain targeting drugs as potential treatment of certain central nervous system diseases,including disorders affecting memory such as Alzheimer’s disease.

Osthole prevents cognitive impairment through modulating neuron cells in Aβ25-35-injected mice

OBJECTIVE To investigate the effects of osthole on learning and memory impairment of AD mice induced by injection of Aβ25-35 and the content of Ca2+, GLU and Aβ1-42 in the brain tissue and peripheral blood. METHODS Mice were randomly assigned to sham operation, Aβ25-35, Aβ25-35+Ost-L,Aβ25-35+Ost-M, and Aβ25-35+Ost-H group. Water maze test was performed to assessing spatial learning ability of mice. It is determined that the MDA level and the activity of SOD in the brain tissue of mice in each group by colorimetry. The GLU kit and Ca2 +kit were used to detect the GLU, Ca2 +in tissue and serum. ELISA was used to detect the expression of Aβ1-42 in the hippocampus and serum of mice. HE staining and silver staining were used to detect neuron apoptosis and pathological changes in brain slices and so on. RESULTS(1) Effects of osthole on learning and memory: With the increase of training day,the escape latencies continuously reduced in each experimental group, the escape latencies of the model group was longer on the 1 st, 2 nd, 3 rd, and 5 thdays than the normal group, the difference was statistically significant(day 3 and 4: P<0.05, day 5: P<0.01);compared with the model group, the escaping latency on the 5 thday of the OST low-medium high-dose group was significantly shortened, which was statistically significant(P<0.05).(2) Effects on oxidative stresspathway: the SOD activity of AD mice in the hippocampus model group was lower than that in the normal group, which was statistically significant(P<0.05);The SOD activity in the OST group was higher than that in the model group, which was statistically significant(P<0.05). The MDA content in the model group was significantly higher than that in the normal group(P<0.05). The MDA content in the OST high-dose group was lower than that in the model group, which was statistically significant(P<0.05).(3) Effects of GLU levels on neurotransmitters:the results of the detection of GLU in cortical area and GLU in serum of AD mice in OST dose groups showed that serum GLU levels in the model group were significantly lower than those in the sham group, which was statistically significant(P<0.05). GLU levels in the cortical area were also significantly higher than those in the sham group, which was statistically significant(P<0.05). Compared with the model group, GLU levels in the OST administration group were significantly down-regulated. Among the serum, the effect of medium dose group was obvious. Although there was a trend of down-regulation in the cortical administration group, there was no statistical significance.(4) Changes in Ca2+concentration in the brain. Detection of intracellular Ca2 +concentration in AD mice by OST doses showed that,compared with the sham group, the model group was significantly upregulated in cortical Ca2 +levels.There was no statistical difference in the administration group. Compared with the model group, the concentration of Ca2+in the OST-H group significantly decreased.(5) Effect on levels of Aβ1-42 in hippocampus and serum: model group had significantly higher Aβ1-42 levels in hippocampus than in sham operation group, which was statistically significant(P<0.05). Aβ1-42 in serum was also significantly upregulated compared to the sham group, which was statistically significant(P<0.05). Compared with the model group, the levels of Aβ1-42 in the OST administration group were significantly down-regulated,with the lower and middle doses in the hippocampus being more significant, while the serum was more pronounced at lower doses.(6) Silver staining to detect the tangles of hippocampal neurons: Neuron tangles in the hippocampal CA1 region showed a dark brown-yellow granule distribution in the nuclei of the model group(positive expression). Nerve cell body and dendrites, axons are black or black red,background light yellow. Compared with the model group, the administration group has improved significantly. CONCLUSION OST improves spatial learning and memory of dementia model mice injected with Aβ25-35 in both hippocampus. Experimental studies have shown that OST has different degrees of regulation on neuronal apoptosis, Ca2 +/GLU/oxidative stress and other pathways, and it plays a role in improving multiple AD pathological changes and delaying the pathogenesis of neurodegenerative diseases.

Effect of osthole,aFructus Cnidii-derivednature coumarin,on osteogenesis and bone healing in mice

OBJECTIVE To investigate the effect of osthole,a natural coumarin isolated from traditional Chinese medicine Fructus Cnidii,on osteogenesis in vitro and bone fracture healing in vivo.METHODS Primary bone marrow mesenchymal stem cells(MSCs)were isolated from 6-week C57/B6 mice,and osteogenic differentiation was assessed by alkaline phosphate(ALP)activity and calcium nodule formation.Adult(12-week)C57mice were subjected to mid-shaft osteotomy on femur.The mice were oral administrated with osthole(5,20 or 50mg·kg-1)or vehicle solvent daily from post-operational week 1.Radiographic imaging,real time molecular imaging,micro computed tomography(μCT)and histology analysis were performed to evaluate the healing progress.RESULTS Results showed that osthole promoted osteogenesis of bone marrow MSCs by enhancing ALP activity and mineralization dose dependently in the range of 1-100μmol·L-1.Plain radiographs showed that administration of osthole at 20 and 50 mg·kg-1 significantly accelerated fracture healing by reducing the period of reparative phase.Further investigation withμCT and histology showed that osthole-treated group had high proportion of newly-formed woven bone and smaller cartilage island compare to control group at week 2;and treatment group had completed endochondral ossification and started remodeling phase at week 3.Molecular imaging of near-infrared(NIR)fluorescent labeled palmidronate depositing on newly formed bone suggested that osthole treatment(20 mg·kg-1)augmented callus mineralization process at both postoperative week 2 and week 3 by 80.72% and 25.95% respectively.CONCLUSION Osthole demonstrates significant osteopromotive effect in vitro and anabolic effect on bone formation in fracture repair,which makes it a potential agent for bone regeneration and against osteoporosis.

Osthole attenuates pulmonary arterial hypertension by modulation of phospholipid metabolism

OBJECTIVE Pulmonary arterial hypertension(PAH)is a malignant pulmonary vascular disease lacking efficacy therapeutics.Therefore,it urgently needs to develop safe and effective drugs for PAH treatment.Osthole derived from Cnidium monnieri(L.)Cusson(Shechuangzi)or Angelica pubescens Maxim(Duhuo)has the capacity to alleviate PAH by decreasing pulmonary arterial pressure and alleviating pulmonary vascular remodeling in rats,which is a candidate drug for the prevention of PAH,but the underlying modulatory mechanism is still unclear.Our study aims at investigating the metabolic modulatory mechanism of osthole against PAH employing functional metabolomics strategy.METHODS PAH model rats were successfully established with MCT,following osthole administration,then functional metabolomics based on untargeted metabolomics assay,targeted lipidomics analysis,qRT-PCR,Western blotting and ELISA were performed to investigate the modulatory mechanism of osthole against pulmonary arterial pressure and pulmonary vascular remodeling in PAH.RESULTS Untargeted metabolomics results found that sphingosine 1-phosphate(S1P)was the differential metabolites characterized PAH and reversed by osthole treatment.S1P is a crucial sphingolipid metabolite catalyzed by sphingosine kinases1(Sphk1)and functions as promoting PASMCs proliferation contributing to pulmonary vascular remodeling and pulmonary arterial pressure increase.We revealed that osthole reversed high level of S1P by modulating metabolic enzyme Sphk1 via inactivating microRNA-21-PI3K/Akt/mTOR signal pathway to decrease pulmonary arterial pressure in rats with PAH.Then,targeted phospholipid metabolomics results uncovered that decadienyl-L-carnitine(C10:2)was the differential metabolite characterized PAH and corrected by osthole treatment in rat with PAH.C10:2 is the intermediate metabolite of fatty acid oxidation(FAO),and C10:2 accumulation indicated mitochondrial dysfunction and FAO increase.CONCLUSION Osthole could block lipid metabolic reprogramming through functional modulating the expression of fatty acid translocase,fatty acid synthase,phospholipase A2,carnitine palmitoyltransferase 1A to inhibit C10:2,thus to improve mitochondrial dysfunction and inhibit utilizing lipid to biosynthesize necessary essence for pulmonary artery smooth muscle cells(PASMCs)proliferation.Moreover,we delineated that C10:2 and metabolic reprogramming enzymes were modulated by miRNA-22-3p which was involved in PASMCs proliferation and pulmonary vascular remodeling.Therefore,osthole inhibited miRNA-22-3p mediated lipid metabolic reprogramming to ameliorate pulmonary vascular remodeling.

Fluorescence spectroscopy of osthole binding to human serum albumin

The interaction of human serum albumin （HSA） with osthole was investigated by fluorescence spectroscopy. Osthole can quench the fluorescence of HSA and the quenching mechanism is a static process. The binding site number n and apparent binding constant K were measured at different temperatures. The thermodynamic parameters △H^0, △G^0 and △S^0 were calculated at different temperatures. The results indicated that electrostatic forces played a major role in the interaction of osthole with HSA. Results of osthole synchronous fluorescence and UV absorption spectra showed that the microenvironment and conformation of HSA were changed.

Compatibility Screening of Plant Extracts Synergistic with Osthole

Plant pests have been a major problem in agricultural production. To establish sustainable methods for plant pest control, 20 of plant material extracts were selected which were synergistic with Osthole to improve the performance of the osthole existing in products. The preliminary screening results indicated that Zanthoxylum bungeanum Maxim.(Z. bungeanum Maxim) compounds exerted the highest synergism. A single dose(2 000 mg · L^(-1)) of Z. bungeanum Maxim. crude extracts was used against Plutella xyllostella(P. xyllostella), Lipaphis erysimi(L. erysimi), and Sitophilus zeamais(S. zeamais). For a further examination form Z. bungeanum Maxim., in which crude extracts were mixed with osthole at different ratios. The volume ratio of 7 : 3(Z. bungeanum Maxim. extract: osthole) exhibited a more pronounced synergistic effect, and all the values of the co-toxicity coefficients were above 120. Eight distinct compounds were derived from the ethanol crude extract from Z. bungeanum Maxim.Then, the toxicity of these components to pests, such as P. xyllostella, L. erysimi, S. zeamais, Rhizopertha dominica(R. dominica),and Tribolium castaneum Herbst(T. castaneum Herbst), was determined. The results indicated that Compound 4 and Compound 7 exerted lethal effects on pest investigation. Compound 4 had the most substantial insecticidal action, at a concentration of64 ug · mL^(-1), and a death ratio of 78.3% was achieved for P. xyllostella.

Research progress on anticancer effects and molecular mechanisms of osthole

Cancer is a serious threat to human life and a big problem in clinical treatment.Some natural active substances extracted from Traditional Chinese Medicine can effectively inhibit the growth of cancer cells,which is a new direction for finding more effective anticancer drugs.Osthole is a natural coumarin compound extracted from Traditional Chinese Medicines such as Cnidium monnieri,Angelica pubescens and Peucedanum praeruptorum Dunn.It has significant inhibitory activity against a variety of cancers.This paper summarizes the anticancer effects and molecular mechanisms of osthole in the treatment of cancers in recent years in order to provide references for further research.

S1-5 Effects of Osthole on Learning and Memory and the Estrogen Pathway in Ovariectomized Rats

Objective:To investigate the effects of osthole,a natural coumarin first derived from the Cnidium plant,on learning and memory,physiological and pathological changes,and expression of estrogen receptor(ER)αandβin the brain of ovariectomized(OVX)rats of Alzheimer’s disease(AD)models.Methods:Female rats were randomly divided into six groups:①sham operation,and OVX plus:②saline,③Estradiol(0.1 mg·kg-1;positive control),④osthole at 12.5 mg·kg-1,⑤osthole at 25 mg·kg-1,and⑥osthole at 50 mg·kg-1;intragastric administration for 30 days.The Morris water-maze test was used to evaluate the learning and memory ability of rats,ELISA to measure the levels of estradiol in the serum,Western blotting to detect the expression of ERαand ERβin the hippocampus,and HE staining to determine the histopathological changes in the brain.Results:①Effects on learning and memory:compared to the OVX alone,osthole at 25 or 50 mg·kg-1 signifi cantly increased the number of entries and the duration in the target quadrant in the water-maze probe trial test(P<0.05).②Effects on the estrogen pathway in the brain:the level of estradiol in the serum and expression of ERβin the hippocampus in the OVX alone were signifi cantly lower,while the expression of ERαwas higher,relative to the sham operation control(P<0.01);osthole at 25 mg·kg-1 reversed the OVX-induced changes in expression of ERαand ERβ(P<0.01).③Effects on histopathological change in the brain:in comparison with the sham operation group,the OVX rats treated with saline displayed increases in the number of apoptotic cells in the hippocampus,which was reversed by osthole at 25 or 50 mg·kg-1(P<0.05),but not the lower dose of 12.5 mg·kg-1.Conclusion:Osthole produced enhancement of learning and memory in the ovariectomized dementia model,which was mediated,at least in part,by regulating neuronal apoptosis and the estrogen pathway.Therefore,osthole is potent in delaying the development of female neurodegenerative diseases,which provides a potential,new approach to treatment of female AD.

NP-14 Effects of Osthole on the Improvement of Learning and Memory Impairment in A Mouse Model Injected with Aβ25-35

Objective:We aimed to investigate the effects of osthole on learning and memory impairment of AD mice induced by injection of Aβ25-35 and the content of Ca2+、GLU、Ab1-42 in the brain tissue and peripheral blood.Methods:Mice were randomly assigned to sham operation,Aβ25-35,Aβ25-35+Ost-L,Aβ25-35+Ost-M,and Aβ25-35+Ost-H group.Water maze test was performed to assessing spatial learning ability of mice.It is determined that the MDA level and the activity of SOD in the brain tissue of mice in each group by colorimetry.The GLU kit and Ca2+kit were used to detect the GLU,Ca2+in tissue and serum.Elisa was used to detect the expression of Aβ1-42 in the hippocampus and serum of mice.HE staining and silver staining were used to detect neuron apoptosis and pathological changes in brain slices.Results:①Effects of osthole on learning and memory:With the increase of training day,the escape latencies continuously reduced in each experimental group,the escape latencies of the model group was longer on the 1st,2nd,3rd,and 5th days than the normal group,the difference was statistically significant(day 3,4:P<0.05,day 5:P<0.01);compared with the model group,the escaping latency on the fifth day of the OST low-medium high-dose group was significantly shortened,which was statistically significant(P<0.05).②Effects on oxidative stresspathway:the SOD activity of AD mice in the hippocampus model group was lower than that in the normal group,which was statistically significant(P<0.05);The SOD activity in the OST group was higher than that in the model group,which was statistically significant(P<0.05).The MDA content in the model group was significantly higher than that in the normal group(P<0.05).The MDA content in the OST high-dose group was lower than that in the model group,which was statistically significant(P<0.05).③Effects of GLU levels on neurotransmitters:the results of the detection of GLU in cortical area and GLU in serum of AD mice in OST dose groups showed that serum GLU levels in the model group were significantly lower than those in the sham group,which was statistically significant(P<0.05).GLU levels in the cortical area were also significantly higher than those in the sham group,which was statistically significant(P<0.05).Compared with the model group,GLU levels in the OST administration group were significantly downregulated.Among the serum,the effect of medium dose group was obvious.Although there was a trend of down-regulation in the cortical administration group,there was no statistical significance.④Changes in Ca2+concentration in the brain;Detection of intracellular Ca ion concentration in AD mice by OST doses showed that,compared with the sham group,the model group was significantly upregulated in cortical Ca2+levels.There was no statistical difference in the administration group.Compared with the model group,the concentration of Ca2+in the OST-H group significantly decreased.⑤Effect on levels of Ab1-42 in hippocampus and serum:model group had significantly higher Ab1-42 levels in hippocampus than in sham operation group,which was statistically significant(P<0.05).Ab1-42 in serum was also significantly upregulated compared to the sham group,which was statistically significant(P<0.05).Compared with the model group,the levels of Aβ1-42 in the OST administration group were significantly down-regulated,with the lower and middle doses in the hippocampus being more significant,while the serum was more pronounced at lower doses.⑥Silver staining to detect the tangles of hippocampal neurons:Neuron tangles in the hippocampal CA1 region showed a dark brown-yellow granule distribution in the nuclei of the model group(positive expression).Nerve cell body and dendrites,axons are black or black red,background light yellow.Compared with the model group,the administration group has improved significantly.Conclusion:OST improves spatial learning and memory of dementia model mice injected with Ab25-35 in both hippocampus.Experimental studies have shown that OST has different degrees of regulation on neuronal apoptosis,Ca2+/GLU/oxidative stress and other pathways,and it plays a role in improving multiple AD pathological changes and delaying the pathogenesis of neurodegenerative diseases.

Preparation of oximoether derivatives of osthole and their pesticidal properties

To explore non-food bioactive products for pesticide candidates,and high value-added application of crop plants in agriculture,by using osthole(a natural coumarin isolated from a Chinese herbal medicine Cnidium monnieri)as a lead compound,a series of osthole derivatives containing oximoether groups were synthesized,and evaluated for their agricultural activities against Tetranychus cinnabarinus and Mythimna separata.The key steric configurations of eight derivatives were confirmed by X-ray crystallography.Especially compound 4r showed potential insecticidal activity(>2-fold of osthole)against M.separata,and 4g displayed good acaricidal activity against T.cinnabarinus(LC_(50)=0.566 mg/mL,>2-fold of osthole).Their structure-activity relationships were also concluded.It will pave the way for future application of osthole derivatives as coumarin-kind botanical pesticides in crop protection.

Osthole ameliorates myonecrosis caused by Clostridium perfringens type A infection in mice

This study aimed to investigate the protective effect of the nature product osthole(OST)against Clostridium perfrin-gens type A infection-caused myonecrosis in a mouse model.Male mice were divided into(1)control,(2)infected,(3)OST50 and(4)OST100 treatment groups.In the infected groups,mice were intramuscularly injected with 1×10^(8) CFU of C.perfringens per day for 6 days.Mice in the OST50 and OST100 groups were administrated intraperitoneally with OST at the doses of 50 or 100 mg/kg per day post C.perfringens infection.Our results showed that C.perfringens infection caused marked necrosis and inflammatory cell infiltration in the muscle tissues of mice.Mice in the OST50 and OST100 treatment groups displayed significantly attenuated C.perfringens infection-induced lipid peroxida-tion,oxidative stress,and apoptosis in their muscle tissue.Furthermore,OST treatment significantly downregulated the expressions of NF-κB,IL-1β,and TNF-αmRNA and protein levels,while concomitantly upregulating the expressions of Nrf2 and HO-1 mRNA and protein.OST treatments also inhibited the expression of phosphorylation(p)-p38,p-mTOR,and p-Erk1/2 proteins,and upregulated LC3II and Beclin1 proteins.In summary,our results reveal that OST therapy confers a protective effect against C.perfringens infection-induced oxidative stress and inflammation in muscle tissue,via activation of Nrf2/HO-1 and autophagy pathways and inhibition of p38,Erk1/2 and NF-κB pathways.

Osthole treatment ameliorates Th2-mediated allergic asthma and exerts immunomodulatory effects on dendritic cell maturation and function

Osthole,an active component of Chinese herbal medicines,reportedly possesses various pharmacological properties and has potential therapeutic applications.This study explored the anti-allergic effects of osthole in asthmatic mice and investigated the immunomodulatory actions of osthole on dendritic cells(DCs)and T cells.Herein,we show that oral administration of osthole to BALB/c mice after ovalbumin(OVA)sensitization ameliorated all of the cardinal features of T helper 2(Th2)-mediated allergic asthma;namely,the production of OVA-specific immunoglobulin E,airway hyperresponsiveness,airway inflammation and the production of Th2-type cytokines including interleukin(IL)-4,IL-5 and IL-13.Surprisingly,IL-10 production was not inhibited and was even enhanced by osthole treatment.We observed a significant increase in the percentages of IL-10-producing DCs and forkhead box P3-positive regulatory T(Treg)cells in osthole-treated asthmatic mice.Additionally,in vitro analyses revealed that osthole-treated bone-marrow-derived DCs had a partial maturation phenotype,secreting large amounts of IL-10 and low levels of proinflammatory cytokines,such as IL-12,IL-6 and tumor necrosis factor-α,and displaying reduced levels of MHC class II surface molecules.These DCs displayed immunosuppressive capacity by directly inhibiting effector T-cell responses or inducing Treg cells.In addition,osthole directly inhibited the activated CD4+T-cell proliferation and Th1/Th2-type cytokine production in this system.Collectively,these results suggest that DCs and T cells are potential target cells responsible for the action of osthole against allergic asthma.

Osthole decreases collagen Ⅰ/III contents and their ratio in TGF-β1-overexpressed mouse cardiac fibroblasts through regulating the TGF-β/Smad signaling pathway

The present study was designed to elucidate whether the mechanism by which osthole decreases collagenⅠ/III contents and their ratio is regulating the TGF-β/Smad signaling pathway in TGF-β1-overexpressed mouse cardiac fibroblasts(CFs). These CFs were cultured and treated with different concentrations of osthole. Our results showed that the TGF-β1 expression in the CFs transfected with that the recombinant expression plasmids pc DNA3.1(+)-TGF-β1 was significantly enhanced. After the CFs were treated with 1.25-5 μg·m L^(-1) of osthole for 24 h, the m RNA and protein expression levels of collagensⅠand III were reduced. The collagen Ⅰ/III ratio was also reduced. The m RNA and protein expression levels of TGF-β1, TβRⅠ, Smad2/3, P-Smad2/3, Smad4, and α-SMA were decreased, whereas the expression level of Smad7 was increased. These effects suggested that osthole could inhibit collagen Ⅰ and III expression and reduce their ratio via the TGF-β/Smad signaling pathway in TGF-β1 overexpressed CFs. These effects of osthole may play beneficial roles in the prevention and treatment of myocardial fibrosis.

Estrogenic activity of osthole and imperatorin in MCF-7 cells and their osteoblastic effects in Saos-2 cells

There is an increasing interest in phytoestrogens due to their potential medical usage in hormone replacement therapy(HRT). The present study was designed to investigate the in vitro effects of estrogen-like activities of two widespread coumarins, osthole and imperatorin, using the MCF-7 cell proliferation assay and their alkaline phosphatase(ALP) activities in osteoblasts Saos-2 cells. The two compounds were found to strongly stimulate the proliferation of MCF-7 cells. The estrogen receptor-regulated ERα, progesterone receptor(PR) and PS2 m RNA levels were increased by treatment with osthole and imperatorin. All these effects were significantly inhibited by the specific estrogen receptor antagonist ICI182, 780. Cell cycle analysis revealed that their proliferation stimulatory effect was associated with a marked increase in the number of MCF-7 cells in S phase, which was similar to that observed with estradiol. It was also observed that they significantly increased ALP activity, which was reversed by ICI182,780. These results suggested that osthole and imperatorin could stimulate osteoblastic activity by displaying estrogenic properties or through the ER pathway. In conclusion, osthole and imperatorin may represent new pharmacological tools for the treatment of osteoporosis.

Synthesis of Osthole Derivatives with Grignard Reagents and Their Larvicidal Activities on Mosquitoes

The structure of osthole has been modified to improve its larvicidal activity against mosquitoes. A new efficient synthesis of osthole derivatives with Grignard reagents has been developed, which employs CuI and LiCI as pro- moters and covers a broad range of substrates to afford the corresponding products in mild to good yields （up to 83%）. Bio-activity evaluation showed that several products exhibited better activities than osthole.

Osthole attenuates pulmonary arterial hypertension by the regulation of sphingosine 1-phosphate in rats

Osthole is observed to have the capacity to treat pulmonary arterial hypertension(PAH) in rats, but molecular mechanism is still unknown. The present study aims to discover therapeutic targets and explore therapeutic mechanism of osthole against PAH from metabolic perspective. A rat model with PAH was successfully established with MCT, following osthole administration, then untargeted metabolomics assay was performed using UPLC-Q-TOF-MS to identify differential metabolites and associated metabolic pathways, at last mechanism investigation was done by qRT-PCR, Western blot and ELISA. Differential metabolites characterized in rats with PAH were mostly assigned to sphingolipid metabolism, synthesis of unsaturated fatty acids, glycolysis, nucleotide metabolism, steroid hormone biosynthesis. Furthermore, osthole reversed high level of S1 P by modulating metabolic enzyme Sphk1 in rats with PAH. In addition, osthole inhibited the expression of Sphk1 by downregulating microRNA-21, phosphorylation of Akt, phosphorylation of mTOR in vivo and in vitro. These results demonstrated that metabolomics is a promising approach to discover potential drug target for PAH treatment. Importantly, our findings further elucidated therapeutic mechanism of osthole, a natural product, having a role of metabolic regulator to potentially treat PAH by targeting inhibition of Sphk1/S1 P via microRNA-21-PI3 K/Akt/mTOR signal pathway. Altogether, this discovery paves a critical foundation for enabling osthole to be a candidate compound to treat PAH.