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.

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.

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 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.

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 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.

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.

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.

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 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.

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.

HPLC法同时测定驻春胶囊中4个成分的含量

目的:建立以高效液相色谱法(HPLC)同时测定驻春胶囊中异补骨脂素、补骨脂素、淫羊藿苷、蛇床子素含量的方法。方法:采用HPLC法,以异补骨脂素、补骨脂素、淫羊藿苷和蛇床子素作为指标成分,采用Agilent ZORBAX SB-C_(18)色谱柱(4.6 mm×250 mm,5μm);流动相为甲醇-水,梯度洗脱,体积流量为1.0 m·min^(-1);柱温为30℃,检测波长为254 nm,进样量为5μL。结果:异补骨脂素、补骨脂素、淫羊藿苷和蛇床子素质量浓度分别在0.0059~0.1770 mg/mL、0.0049~0.1470 mg/mL、0.0031~0.093 mg/mL和0.0048~0.1440 mg/mL范围内与峰面积呈良好的线性关系,其相关系数分别为0.9996、0.9995、0.9997和0.9995,平均加样回收率分别为101.45%、100.95%、100.46%和101.17%;RSD分别为2.19%、1.83%、1.71%和2.01%。结论:该分析方法适用于驻春胶囊中异补骨脂素、补骨脂素、淫羊藿苷和蛇床子素的含量测定。

蛇床子素化学结构修饰研究进展

蛇床子素,其化学名为7-甲氧基-8-异戊烯基香豆素,又名甲氧基欧芹酚,是蛇床子成熟果实中的主要有效成分,属香豆素类化合物,拥有丰富的生物活性。为深入研究蛇床子素活性与其结构的关系,人们对其进行了大量的结构修饰研究工作,修饰部位主要集中在内酯环、C-7位甲氧基和C-8位异戊烯基。综述蛇床子素的结构修饰研究进展,以期为该类化合物结构修饰深入研究和应用开发提供参考。

Osthol和Icarin对甲减小鼠血清甲状腺激素的影响

目的：观察蛇床子素（Ｏｓｔｈｏｌ）和淫羊藿甙（Ｉｃａｒｉｎ）对甲减“肾阳虚”小鼠血清甲状腺激素的影响。方法：用丙基硫氧嘧啶（ＰＴＵ）制备甲减“肾阳虚”小鼠模型，用放射免疫法测定小鼠血清Ｔ３、ｒＴ３和Ｔ４的浓度。结果：模型鼠血清Ｔ３、ｒＴ３和Ｔ４浓度皆降低（Ｐ＜０．０１），死亡率升高（Ｐ＜０．０１），造模过程中同时分别给药Ｏｓｔｈｏｌ和Ｉｃａｒｉｎ能有效地阻止Ｔ３、ｒＴ３和Ｔ４浓度下降（Ｐ＜０．０１或Ｐ＜０．０５），死亡率也显著降低（Ｐ＜０．０１）。同时模型鼠分别经Ｏｓｔｈｏｌ和Ｉｃａｒｉｎ治疗后，Ｔ３、ｒＴ３和Ｔ４浓度得到明显提高（Ｐ＜０．０１或Ｐ＜０．０５）。结论：Ｏｓｔｈｏｌ和Ｉｃａｒｉｎ既能拮抗ＰＴＵ的甲状腺抑制作用，预防“肾阳虚”证的出现，又能促进ＰＴＵ所致甲减“肾阳虚”小鼠体内甲状腺激素水平的提高，具有补肾壮阳作用。

蛇床子素通过减轻阿尔茨海默病大鼠模型中的神经炎症改善认知功能障碍

目的抗炎治疗为阿尔茨海默病(AD)的治疗提供了一个潜在的靶点。蛇床子素(Ost)已被证明可以改善记忆并具有多效性保护作用,但其在AD治疗过程中的确切机制尚不清楚。方法在本研究中,作者研究Ost对β-淀粉样蛋白1-42(Aβ1-42)诱导的认知功能障碍和海马炎症反应的保护作用。采用双侧海马注射Aβ_(1-42)建立AD大鼠模型,在Aβ_(1-42)注射前7 d至注射后7 d每日给予Ost。采用Morris水迷宫(MWM)试验评估大鼠的认知功能。采用免疫组化和Western blot法检测海马神经胶质反应及Toll样受体4(TLR4)、肿瘤坏死因子受体相关因子6(TRAF6)和核因子-κB(NF-κB)的表达。用酶联免疫吸附试验(ELISA)测定促炎细胞因子的产生。结果Ost治疗显著改善了AD大鼠模型海马中Aβ_(1-42)毒性诱导的认知障碍,减轻了胶质细胞的激活,降低了炎症细胞因子的释放(P<0.01)。经Ost处理后,Aβ_(1-42)诱导的TLR4、TRAF6和NF-κB表达的明显降低(P<0.01)。结论Ost通过抑制TLR4/NF-κB信号通路而减轻Aβ_(1-42)诱导的神经炎症,可能是一种很有前途的治疗药物。

蛇床子素调节PI3K/Akt/m TOR信号通路对老龄自发性高血压大鼠肾氧化应激损伤的影响

目的:探究蛇床子素对老龄自发性高血压大鼠的影响机制。方法:购买20月龄的老龄自发性高血压大鼠(SHRs)及健康Wistar-Kyoto(WKY)大鼠,对SHRs进行蛇床子素灌胃治疗处理持续8周。对大鼠收缩压及舒张压进行监测,使用苏木精-伊红(H&E)、碘酸雪夫染色(PAS)和Masson染色观察大鼠肾脏组织病理学变化。使用ELISA试剂盒检测大鼠肾脏超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)活性。采用Western blot对PI3K/Akt/mTOR信号通路相关蛋白进行检测。结果:蛇床子素降低SHRs收缩压及舒张压,改善了SHRs大鼠肾脏的组织病理学变化,降低SHRs肾脏MDA活性,提升SOD及GSH活性。蛇床子素降低p-PI3K、p-Akt、p-mTOR水平。结论:蛇床子素通过降低PI3K/Akt/mTOR信号通路活性发挥对老龄自发性高血压大鼠肾氧化应激损伤的保护作用。

蛇床子素通过NF-kB信号通路调控退变髓核细胞的细胞外基质表达、炎症反应及凋亡

目的 研究蛇床子素对退变髓核细胞的细胞外基质表达、炎症反应和凋亡的作用及其潜在机制。方法 采用氧糖剥夺(Oxygen Glucose Deprivation, OGD)培养建立退变髓核细胞模型,将细胞分为:对照组(正常培养)、OGD组(OGD培养)、蛇床子素组(OGD培养+蛇床子素处理)、PDTC组(OGD培养+NF-κB通路抑制剂PDTC处理)和蛇床子素+PDTC组(OGD培养+蛇床子素+PDTC处理)。采用Western blot检测细胞中Ⅱ型胶原(Collagen typeⅡ,Col2al)、聚集蛋白聚糖(Aggrecan)、P65和p-P65蛋白表达,ELISA法检测细胞培养液中炎性因子血清白细胞介素-1β(interleukin-1 β,IL-1β),白细胞介素-6(interleukin-6,IL-6)和肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)的水平,流式细胞仪检测细胞凋亡率。结果 与对照组比较,OGD组细胞中Col2al、Aggrecan蛋白相对表达水平明显降低,细胞培养液中IL-1β、IL-6和TNF-α水平升高,细胞凋亡率明显升高,细胞中p-P65蛋白表达明显升高;与OGD组比较,蛇床子素组和PDTC组细胞中Col2al、Aggrecan蛋白相对表达水平明显升高,细胞培养液中IL-1β、IL-6和TNF-α水平降低,细胞凋亡率明显降低,细胞中p-P65蛋白表达明显降低,而蛇床子素+PDTC组上述指标改变最为明显。结论 蛇床子素可能通过抑制核因子激活的B细胞的κ轻链增强(nuclear factorκB,NF-κB)信号通路的活性,促进退变髓核细胞的细胞外基质合成、抑制退变髓核细胞的炎症反应和凋亡。

蛇床子对肾阳虚骨质疏松大鼠的治疗作用与机制研究

[目的]探索蛇床子对肾阳虚骨质疏松大鼠的骨保护作用与分子机制。[方法]SD大鼠分为正常、模型、蛇床子提取物与蛇床子素组。除正常组外,其余大鼠每日皮下给予氢化可的松,给药组每日灌胃给药,连续3周。大鼠麻醉后采集股骨,其中左侧股骨进行抗酒石酸磷酸酶染色、骨钙素与转化生长因子-β(transforming growth factor-β,TGF-β)免疫组化染色以及骨密度检测,右侧股骨进行实时荧光定量聚合酶链式反应与免疫印迹检测靶基因和蛋白表达。[结果]氢化可的松可显著促进大鼠破骨细胞分化与成熟,抑制成骨细胞生成,降低骨小梁生成,降低骨密度,最终诱导大鼠肾阳虚骨质疏松发生。生物信息预测结果提示,TGF-β信号通路是蛇床子潜在候选靶标通路。蛇床子可显著减轻由氢化可的松诱导的破骨细胞分化与成熟现象,并促进成骨细胞分化与成熟,从而减轻氢化可的松诱发的大鼠骨质疏松。蛇床子提取物可显著增强股骨中因氢化可的松诱导的TGF-β基因下调。[结论]蛇床子可通过调控TGF-β信号通路,抑制肾阳虚大鼠骨质疏松。