香青兰化学成分研究

目的:研究香青兰化学成分。方法:采用活性追踪分离的方法,对香青兰全草进行了系统萃取,反复柱层析。结果:从总浸膏中分离得到了1个化合物,命名为α,从石油醚层和氯仿层分别得到3个化合物,分别命名为S2、S3和L1。4个化合物鉴定为:化合物α为田蓟甘(tilianin),化合物L1为熊果酸(ursolie acid),化合物S2为豆甾醇(stigmasterol),化合物S3为乌发醇(uvqol)。结论:化合物S2、S3和化合物L1均为该植物首次获得。

A New Stigmasterol and a New Eremophilenolide from Ligularia dolichobotrys

A new stigmasterol 3, 7, 22-trihydoxystigmast-5-ene (1) and a new eremophilen- olide 8-methoxy-6-angeloyloxyeremophil-7(11)-en-8, 12-olide-14-oic acid (2) were isolated from Ligularia dolichobotrys Diels. Their structures were deduced on the basis of spectral data.

In vivo combination therapy of chloroquine and stigmasterol against Plasmodium berghei induced pathologies in mice

Objective:To investigate the effect of combination therapy of chloroquine and stigmasterol on Plasmodium berghei(thereafter referred to as P.berghei)malaria-induced organ pathologies.Methods:Thirty five mice weighing 20-30 g were placed into seven groups of five mice each and distributed as uninfected administered 100 mg/kg BW stigmasterol,uninfected administered only feed and water ad libitum,infected with P.berghei and-administered 50 mg/kg BW stigmasterol,100 mg/kg BW stigmasterol,100 mg/kg BW stigmasterol plus 5 mg/kg BW chloroquine,and 5 mg/kg BW chloroquine.The last group of mice served as P.berghei infected and not treated control.The levels of parasitemia,packed cell volume,and other biochemical parameters were measured.Results:Combination therapy of P.berghei infection with stigmasterol and chloroquine did not significantly(P>0.05)reduce parasitemia level while stigmasterol treatment alone significantly(P<0.05)reduced the parasitemia level.However,the P.berghei induced anemia was decreased significantly(P<0.05)upon treatment with a combination of chloroquine and stigmasterol as well as with stigmasterol alone.Furthermore,the combination of chloroquine and stigmasterol significantly(P<0.05)decreased the activities of serum alanine aminotransferase,aspartate aminotransferase,urea and spleen total proteins levels in P.berghei mice in comparison with the untreated group.Treatment of P.berghei infected mice with stigmasterol alone and in combination with chloroquine significantly(P<0.05)increased the level of serum creatinine while serum and spleen malondialdehyde levels were significantly(P<0.05)decreased.Levels of glutathione in spleen and kidney were insignificantly(P>0.05)altered upon treatment with both doses of stigmasterol as well as the combination therapy.Conclusions:This study concluded that the combination of stigmasterol and chloroquine could combat anemia and some organ pathologies associated with P.berghei infection.

In vivo combination therapy of chloroquine and stigmasterol against Plasmodium berghei induced pathologies in mice

Objective:To investigate the effect of combination therapy of chloroquine and stigmasterol on Plasmodium(P.)berghei malaria-induced organ pathologies.Methods:Totally 35 mice weighing 20-30g were placed into 7 groups of 5 mice each and distributed as uninfected administered 100 mg/kg BW stigmasterol,uninfected administered only feed and water ad libitum,infected with P.berghei and administered 50 mg/Kg BW stigmasterol,100 mg/kg BW stigmasterol,100 mg/kg BW stigmasterol plus 5 mg/kg BW chloroquine,and 5 mg/kg BW chloroquine.The last group of mice served as P.berghei infected and not treated control.The levels of parasitemia,packed cell volume,and other biochemical parameters were measured.Results:Combination therapy of P.berghei infection with stigmasterol and chloroquine did not significantly(P>0.05)reduce parasitemia level while stigmasterol treatment alone significantly(P<0.05)reduced the parasitemia level.However,the P.berghei induced anemia was decreased significantly(P<0.05)upon treatment with a combination of chloroquine and stigmasterol as well as with stigmasterol alone.Furthermore,the combination of chloroquine and stigmasterol significantly(P<0.05)decreased the activities of serum alanine aminotransferase,aspartate aminotransferase,urea and level of spleen total proteins in P.berghei infected mice in comparison with the untreated group.Treatment of P.berghei infected mice with stigmasterol alone and in combination with chloroquine significantly(P<0.05)increased the level of serum creatinine while serum and spleen malondialdehyde levels were significantly(P<0.05)decreased.Levels of glutathione in spleen and kidney were insignificantly(P>0.05)altered upon treatment with both doses of stigmasterol as well as the combination therapy.Conclusions:This study concluded that the combination of stigmasterol and chloroquine could combat anemia and some organ pathologies associated with P.berghei infection.

Mechanism of Shenfu Decoction in the treatment of critically ill patients with Coronavirus Disease 2019 (COVID-19) based on network pharmacology

Objective:The purpose of this thesis is to explore the mechanism of ShenFu Decoction in the treatment of critically ill patients with COVID-19 based on network pharmacology.Methods:The primary active ingredients and potential targets of ShenFu Decoction were searched from the TCMSP database.The targets of COVID-19 were obtained by searching the GeneCards and OMIM databases.A ShenFu Decoction-compound-target-COVID19 network and a protein-protein interaction(PPI)network were respectively constructed through the Cytoscape 3.5.1 software and the STRING database.Gene Ontology(GO)function enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis were performed via Bioconductor bioinformatics software package and R programming language.Results:ShenFu Decoction contains 255 compounds and 94 potential targets.43 primary active ingredients were searched from the TCMSP database with oral bioavailability(OB)≥30%and drug-likeness(DL)≥0.18 as the retrieval condition.Numbers of targets of COVID-19 were 352 by searching the GeneCards and the OMIM databases.16 key targets were acquired by intersecting the targets of drug with the targets of disease.There were 49 GO terms and 102 pathways after analyzing GO and KEGG.Conclusion:Kaempferol,ginsenoside rh2,beta-sitosterol,Stigmasterol and Deoxy andrographolide might be the main active ingredients which may cause the inhibition of the SARS-CoV-23CL hydrolase activity and regulate ACE2.As a result,the antiviral effect,immunoregulation,targeting cytokine storm of SFD may play an important role in the treatment of critically ill patients with COVID-19 through regulating multiple signaling pathways such as AGE-RAGE signaling pathway in diabetic complications,IL-17 signaling pathway,C-type lectin receptor signaling pathway,HIF-1 signaling pathway.

Determination of phytosterols in <i>Oryza sativa</i>L. cultivars by liquid chromatography atmospheric chemical ionization time-of-flight mass spectrometer (LC-APCI-TOF-MS) coupled with a modified QuEChERS method

Development of analytical methods for the determination of phytosterols in whole grains is one of growing interests due to their clinical and nutritional functions in human. In this study, we examined simultaneous determination of β-si tosterol, stigmasterol and campesterol in Korean rice cultivars by liquid chromatography atmospheric chemical ionization time-of-flight mass spectrometer (LC-APCI-TOF-MS). The hulled rice samples were extracted using a modified QuEChERS and analyzed by LC-APCI -TOF-MS. The method was validated by low limit of detection (LOD) and limit of quantification (LOQ) values and high recovery values with low relative standard deviation (RSD). The highest concentration of sitosterol was detected in Hwasung and Undu rice cultivars, exhibiting approximately 195.5 and 191.7 mg/Kg, respectively. Stigmasterol and campesterol were detected at the highest level in Hwasung rice cultivar, giving the concentrations of approximately 70.0 and 53.9 mg/Kg, respectively. Sitosterol was determined to be a main phytosterol detected in the rice samples. Our data demonstrated that LC/APCI-TOF- MS coupled with QuEChERS method can be used as a simple tool for the determination of phytosterols in rice samples.

Cytotoxic, Antimicrobial and Antioxidant Properties of <i>Commelina diffusa</i>Burm. F.

Commelina diffusa Burm. F. is a herbaceous tropical plant with different traditional medicinal uses. Present study is aimed to isolate the bioactive compounds from DCM-Methanol extract of the powdered whole plant of Commelina diffusa and to investigate the cytotoxic, antimicrobial and antioxidant activities of the crude extract and its twelve vacuum liquid chromatographic fraction (CD1-12). Vacuum Liquid Chromatography (VLC) was employed to isolate bioactive metabolites. The physicochemical properties of the isolated compound were examined and the molecular structure was elucidated by NMR spectroscopy.?Cytotoxic, antibacterial, antioxidant activities were evaluated following brine shrimp lethality bioassay, disc diffusion method and DPPH free radical scavenging assay respectively. The isolated compound was identified as steroid (stigmasterol) which had significant cytotoxic effect on vero cell line. The crude extract and its fractions (CD8-CD12) exhibited strong cytotoxicity in brine shrimp lethality bioassay having LC50 values 3.79, 9.19, 29.49, 16.60, 19.36, 44.58 μg/mL respectively. The crude extract showed mild to strong antibacterial activity. Fractions (CD5-CD12) showed mild to strong antibacterial activity against Escherichia coli in comparison with Kanamycin (standard). Strong antioxidant activities were found in crude extract (IC50 value 30.52 μg/ml), CD11 (IC50 value 39.27 μg/ml) and CD12 (IC50 value 19.50 μg/ml). The study suggests Commelina diffusa plant extract to have strong antioxidant and cytotoxic activity which is indicative of presence of compounds with broad spectrum of curative applications. One compound namely stigmasterol has been isolated from the plant.

Stigmasterol protects human brain microvessel endothelial cells against ischemia-reperfusion injury through suppressing EPHA2 phosphorylation

Stigmasterol is a plant sterol with anti-apoptotic,anti-oxidative and anti-inflammatory effect through multiple mechanisms.In this study,we further assessed whether it exerts protective effect on human brain microvessel endothelial cells(HBMECs)against ischemia-reperfusion injury and explored the underlying mechanisms.HBMECs were used to establish an in vitro oxygen and glucose deprivation/reperfusion(OGD/R)model,while a middle cerebral artery occlusion(MCAO)model of rats were constructed.The interaction between stigmasterol and EPHA2 was detected by surface plasmon resonance(SPR)and cellular thermal shift assay(CETSA).The results showed that 10μmol·L−1 stigmasterol significantly protected cell viability,alleviated the loss of tight junction proteins and attenuated the blood-brain barrier(BBB)damage induced by OGD/R in the in vitro model.Subsequent molecular docking showed that stigmasterol might interact with EPHA2 at multiple sites,including T692,a critical gatekeep residue of this receptor.Exogenous ephrin-A1(an EPHA2 ligand)exacerbated OGD/R-induced EPHA2 phosphorylation at S897,facilitated ZO-1/claudin-5 loss,and promoted BBB leakage in vitro,which were significantly attenuated after stigmasterol treatment.The rat MCAO model confirmed these protective effects in vivo.In summary,these findings suggest that stigmasterol protects HBMECs against ischemia-reperfusion injury by maintaining cell viability,reducing the loss of tight junction proteins,and attenuating the BBB damage.These protective effects are at least meditated by its interaction with EPHA2 and inhibitory effect on EPHA2 phosphorylation.

Semi-synthesis of Several Stigmasterol Saponins

Several stigamasterol saponins were concisely synthesized. Name!y, four monosaccharide （glucopyranose, galactopyranose, xylopyranose, 2-acetamido-2-deoxy-a-D-glucopyranose）, lactopyranose and chacotriose were coupled with 3-OH of stigmasterol. All the compounds were identified by NMR, IR and high resolusion MS.

A New Stigmasterol Ester from Aeschynomene indica

Objective To study the chemical constituents of Aeschynomene indica.Methods The constituents were isolated and purified by means of silica gel column chromatography and recrytallization,and the structures were elucidated by physicochemical properties and spectral analyses.Results Twelve compounds were obtained and elucidated as stigmasterol tritriacontanate(1),monotetracontane(2),taraxerol(3),stigmasterol(4),stearic acid(5),heptatria- contanoic acid(6),arachidic acid(7),ursolic acid acetate(8),quercetin(9),myricetin(10),myricetin-3-O-rhamnoside(11),and rutoside(12).Conclusion All the compounds are isolated from this plant for the first time and compound 1 is a new one.

Effect of Ipomea carnea Jacq. flowers on hematological changes in toluene diisocyanate-induced inflammation in Wistar rats

AIM： To investigate the active chloroform fraction of the ethanol extract oflpomoea carnea flowers on hematological changes in toluene diisocyanate-induced inflammation in Wistar rats. METHOD： Except for the control group, all of the rats were sensitized with intranasal application of 5 μL of 10% toluene diiso- cyanate （TDI） for 7 days. One week after second sensitization, all of the rats were provoked with 5 ~tL of 5% TDI to induce airway hypersensitivity. After the last challenge, blood and bronchoalvelor lavage （BAL） fluid were collected and subjected to total and differential leucocytes count. Flash chromatography was performed on the most active chloroform fraction to isolate an individual component. RESULTS： Treatment with the ethanolic extract and its chloroform fraction at an oral dose of 200 mg·kg^ -1 showed a significant decrease in circulating neutrophil and eosinophil in blood and BAL as compared with standard dexamethasone （DEXA）. The structure of the compound obtained from chloroform fraction oflpomea carnea was elucidated as stigmast-5, 22-dien-3β-ol on the basis of spectral data analysis. CONCLUSION： The chloroform fraction was found to be more effective to suppress airway hyper reactivity symptoms, and de- creased count of both total and differential inflammatory cells.

Phytochemical investigation and simultaneous estimation of bioactive lupeol and stigmasterol in Abutilon indicum by validated HPTLC method

Objective:To perform a simultaneous quantitative estimation of two biologically active triterpenoid compounds lupeol and a steroid compound,stigmasterol,in Abutilon indicum(A.indicum)using high-performance thin-layer chromatography(HPTLC).Methods:TLC aluminum plates precoated with silica-gel 60 F254(20 cmí10 cm)were used with a mobile phase of toluene-methanol-formic acid(7.0:2.7:0.3,v/v/v)and densitometric determination of these compounds was carried out at 530 nm in reflectance/absorbance mode.Results:Compact bands for lupeol and stigmasterol were obtained at R_(f)0.52±0.02 and 0.28±0.05.The limit of detection(45 and 18 ng/band),limit of quantification(135 and 54 ng/band),recovery(98.2%-99.7%and 97.2%-99.6%)and precision(≤2.18 and 1.91)were satisfactory for lupeol and stigmasterol respectively.Linearity range for lupeol and stigmasterol were 100-1000(r^(2)=0.9994)and 50-500 ng/band(r^(2)=0.9941)and the contents were estimated as(0.59±0.10)%and(0.83±0.10)%w/w respectively.The total phenolic,flavonoid,proanthocyanidin,alkaloidal and saponin contents of methanolic extract of A.indicum were also measured in this work.According to International Conference on Harmonization(ICH)guidelines,the method was validated for linearity,precision,accuracy,and recovery,limit of detection,limit of quantification,specificity,and robustness.Conclusions:The HPTLC method was found to be reproducible,accurate,and precise and could detect these two compounds at nanogram level from the A.indicum.