CN105232539B - Two breeds of horses bitterroot source organism alkali is used as the application for preparing anti-inflammatory drug - Google Patents
Two breeds of horses bitterroot source organism alkali is used as the application for preparing anti-inflammatory drug Download PDFInfo
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Abstract
Two breeds of horses bitterroot source organism alkali is as the application for preparing anti-inflammatory drug or health products, and described two purslane source organism alkali are oleracimine and oleracone.The anti-inflammatory refers to suppress the inflammation caused by the excessive inflammatory cytokine of macrophage release or inflammatory mediator.The present invention is tested by the suppression to LPS inducing mouse macrophage RAW264.7 inflammatory reactions, shows that purslane source oleracimine and oleracone has suppression inflammatory factor IL 6, TNF α and suppresses inflammatory mediator NO, PGE2Effect, detected by Western blot and real-time fluorescence quantitative PCR show that oleracimine is inhibited to the albumen of iNOS, COX 2 and gene expression, and valid certificates two breeds of horses bitterroot source organism alkali has effects that anti-inflammatory.To sum up, purslane source organism alkali oleracimine and oleracone have good anti-inflammatory activity, can be used to prepare anti-inflammatory activity medicine as lead compound, to prevent or treat by NO, IL 6, TNF α and PGE2Inflammation caused by excess.
Description
Technical field
It is applied to prepare answering for anti-inflammatory drug the present invention relates to biomedicine field, more particularly to purslane source organism alkali
With.
Background technology
Purslane also known as long life dish, horse three-coloured amaranth, belong to the herb of the annual meat herbaceous plant purslane of Portulacaceae.Dent
Shade tolerant that amaranth is fast light, happiness is warm and drought-enduring waterlogging, and suitable growth is being air-dried and the place of soil moisture.It is purslane growing power, anti-
Sick ability is strong, and widely distributed, aboundresources, as one of integration of drinking and medicinal herbs wild plant, is both a kind of health functional food, has again
There is important medical value.2015 editions《Pharmacopoeia of People's Republic of China》In record the dry aerial parts of purslane and be used as medicine, have
The effects such as having clearing heat and detoxicating, cooling blood and hemostasis, stop dysentery, available for toxic-heat and blood stasis, carbuncle swells furunculosis, eczema, erysipelas, snake bite and insect sting, just
Blood, hemorrhoid blood and metrostaxis.Research shows that the pharmacological action various with it of contained a variety of chemical compositions is closely bound up in purslane,
Its main chemical compositions includes：Organic acid, flavonoids, terpene, Coumarins, alkaloids, volatile oil and polysaccharide etc..It is modern
Pharmaceutical research shows that purslane has reducing blood lipid, hypoglycemic, anti-inflammatory, anti-oxidant, antitumor, antiatherosclerosis, relaxation
Or the effects such as excited smooth muscle and strengthen immunity.
In recent years, purslane enjoys scholar because of the characteristic of its abundant nutrition, various pharmacological action and medicine-food two-purpose
Concern.Up to the present, the research on purslane anti-inflammatory activity concentrates on the report of purslane different solvents extract, this group
Research finds that purslane source organism alkali has anti-inflammatory efficacy, may be the main compound of its antiinflammatory action.
Inflammation is a kind of defense reaction of the body for stimulation, often shows as red, swollen, hot, pain and other dysfunctions.When
When body is stimulated by foreign pathogen, often cause various inflammatory mediators excessively release and inflammatory cell excessive activation.Macrophage
The important pass that cell is attacked as fight exotic, can adjust body defenses function, can secrete inflammatory factor and inflammation
Medium, including TGF（Transforming growth factor, TGF）, interleukins（Interleukin,
IL）, TNF（Tumor necrosis factor, TNF）, prostaglandin 2（Prostaglandin E2, PGE2）With
And nitric oxide（Nitric oxide, NO）Deng, these bioactive substances can also influence the expression of enzyme during inflammatory reaction,
Such as inducible nitric oxide synzyme（Inducible nitric oxide synthase, iNOS）And Cycloxygenase
（Cyclooxygenase 2, COX-2）.
Bacteria lipopolysaccharide（Lipopolysaccharides, LPS）It is a kind of composition in gram-negative bacterial cell wall,
It forms external inflammatory model after invading macrophage as pathogen, and macrophage can be made largely to secrete above-mentioned biological active matter
Matter.
The content of the invention
The application for preparing anti-inflammatory drug is used as it is an object of the invention to provide two breeds of horses bitterroot source organism alkali.
To realize the above-mentioned purpose of the present invention, the present invention provides two breeds of horses bitterroot source organism alkali as preparing anti-inflammatory drug
Application, described two purslane source organism alkali be oleracimine and oleracone.
The molecular formula of the oleracimine is C18H26N2O, structural formula is.
The molecular formula of the oleracone is C14H17NO2, structural formula is.
Compound and its salt or derivative containing oleracimine in the anti-inflammatory drug；Contain in the anti-inflammatory drug
There are oleracone compound and its salt or derivative.
The anti-inflammatory refers to suppress the inflammation caused by the excessive inflammatory cytokine of macrophage release or inflammatory mediator；
The inflammatory cytokine includes IL-6, TNF-α, and the inflammatory mediator is NO, PGE2。
The anti-inflammatory drug is that inflammatory cytokine over-expresses the inhibitor that inhibitor, macrophage produce excessive NO
Deng.
Beneficial effects of the present invention compared with prior art.
The two breeds of horses bitterroot source organism alkali that the present invention is provided is as the application for preparing anti-inflammatory drug, by inducing LPS
The suppression experiment of mouse macrophage RAW264.7 inflammatory reactions, shows purslane source oleracimine and oleracone tools
There are suppression inflammatory factor IL-6, TNF-α and suppress inflammatory mediator NO, PGE2Effect, detected by Western blot and real-time fluorescence
Quantitative PCR shows that oleracimine is to iNOS, COX-2 albumen and gene expression inhibited, valid certificates two breeds of horses
Bitterroot source organism alkali has effects that anti-inflammatory.To sum up, purslane source organism alkali oleracimine and oleracone have
Have good anti-inflammatory activity, can as lead compound be used for prepare anti-inflammatory activity medicine, come prevent or treat by NO, IL-6,
TNF-α and PGE2Inflammation caused by excess.
Brief description of the drawings
Figure 1A is the influence schematic diagram for the survival rate that oleracimine induces LPS RAW264.7 macrophages.
Figure 1B is the influence schematic diagram for the survival rate that oleracone induces LPS RAW264.7 macrophages.
Fig. 2A is that oleracimine stimulates LPS RAW264.7 cells generation NO inhibitory action schematic diagram.
Fig. 2 B are that oleracone stimulates LPS RAW264.7 cells generation NO inhibitory action schematic diagram.
Fig. 3 A are that oleracimine stimulates LPS RAW264.7 cells secretion IL-6 influence schematic diagram.
Fig. 3 B are that oleracone stimulates LPS RAW264.7 cells secretion IL-6 influence schematic diagram.
Fig. 4 A are that oleracimine stimulates LPS RAW264.7 cell TNF secretions-α influence schematic diagram.
Fig. 4 B are that oleracone stimulates LPS RAW264.7 cell TNF secretions-α influence schematic diagram.
Fig. 5 A are that oleracimine stimulates RAW264.7 cells to secrete PGE LPS2Influence schematic diagram.
Fig. 5 B are that oleracone stimulates RAW264.7 cells to secrete PGE LPS2Influence schematic diagram.
Fig. 6 A are the influence schematic diagram for the iNOS albumen that oleracimine stimulates LPS the expression of RAW264.7 cells.
Fig. 6 B are the influence schematic diagram for the COX-2 albumen that oleracimine stimulates LPS the expression of RAW264.7 cells.
Fig. 6 C are that oleracimine stimulates LPS the protein band figure of RAW264.7 cells expression.
Fig. 7 A are that oleracimine stimulates LPS the iNOS effect gene schematic diagrames of RAW264.7 cells expression.
Fig. 7 B are that oleracimine stimulates LPS the COX-2 effect gene schematic diagrames of RAW264.7 cells expression.
Embodiment
The present invention is further described with reference to specific embodiment.
The present embodiment provides two breeds of horses bitterroot source organism alkali and is used as the application for preparing anti-inflammatory drug, described two purslanes
Source organism alkali is oleracimine and oleracone.
For further checking beneficial effects of the present invention, there is provided following experimental example.
1st, experimental raw and related reagent, packet.
DMEM high glucose mediums, hyclone are purchased from Hyclone companies of the U.S.；Penicillin, streptomysin are purchased from the Hangzhou four seasons
Blue or green company；LPS is purchased from Sigma Co., USA；IL-6、TNF-α、PGE2ELISA kit be purchased from Cayman companies of the U.S.；Carefully
Cellular lysate liquid, PMSF and Griess reagents are purchased from green skies Bioisystech Co., Ltd；BCA determination of protein concentration kit,
Primer is purchased from Shanghai Sheng Gong companies；INOS monoclonal antibodies, COX-2 monoclonal antibodies, internal reference monoclonal antibody are purchased from U.S. CST
Company；Two antiantibodys of HRPO mark are purchased from Beijing biotech firm of Zhong Shan Golden Bridge；Pvdf membrane is purchased from the U.S.
Millipore companies；Chemical luminous substrate（ECL）It is purchased from Pierce companies of the U.S.；EastepTMTotal RNA extraction reagent box,
GoScriptTMReverse transcription reagent box is purchased from Promega companies；SYBR Premix ExTaq TMKit is purchased from TaKaRa companies.
Packet：It is divided into three groups, i.e. control group, LPS groups and experimental group.
2nd, two breeds of horses bitterroot source organism alkali is oleracimine and oleracone preparation method.
（1）Purslane source organism alkali is oleracimine preparation method, is comprised the following steps.
Step 1, weigh purslane and dry medicinal material 50kg, using water boiling and extraction, the consumption of water is the 8-16 of medicinal material consumption
Times, decoct and extract twice, each 2h, Aqueous extracts filtering, merging filtrate is directly concentrated, and obtains decoction standby.
Step 2, by step 1 gained decoction directly preprocessed mistake AB-8 or D101 macroporous absorbent resins absorption remove
It is miscellaneous, respectively with water and 50% ethanol elution, 50% ethanol eluate is concentrated under reduced pressure, concentrate is obtained standby.The water
It is 2-3 column volume to elute removal of impurities consumption, and the elution consumption of 50% ethanol is to rush untill lighter.The macropore is inhaled
The preprocessing process of attached resin be ethanol soaked 24 hours, upper prop, with ethanol be washed till instillation water in without muddiness, be washed with water to
Without alcohol taste.
Step 3, by concentrate in step 2 with ethyl acetate repeatedly 3 times extraction, less than 40 DEG C are recovered under reduced pressure ethyl acetate extremely
Medicinal extract, obtains acetic acid ethyl ester extract.The amount ratio of the ethyl acetate and concentrate is 1:1（v:v）.
Step 4：By acetic acid ethyl ester extract dry method loading in step 3, separated through silica gel column chromatography, wherein silica gel is with 200-
400 mesh, successively with petroleum ether-acetone（1:1、1:2、1:3、1:5, v:v）Gradient elution, each ratio elutes to obtain 40 positions
（I.e. each ratio elutes to obtain 40 bottles, every bottle of 200mL）, 160 positions are obtained（160 bottles are obtained）, through thin layer color
Spectrum is detected, is developed the color, and merges the 90-130 elutions position of colour developing（Merge the 90-130 bottles of colour developing, discard 1-89 bottles with
131-160 bottles）, it is dry by being concentrated under reduced pressure into below 40 DEG C of 90-130 positions after merging, it is standby.
Step 5：Gains in step 4 are separated through ODS medium pressure column chromatographies again（Octadecylsilyl, octadecyl silicon
Alkane bonded silica gel filler, filler particle size is 20-40 μm）, use methanol-water（50/50, v/v）Isocratic elution（Pressurization, makes the flow velocity be
1ml/min, temperature is room temperature）, obtain 10 positions（I.e. isocratic elution obtains 10 bottles, and every bottle of 100mL is marked successively）, through thin
Layer chromatography detected, is developed the color, and merges 6-7 positions, less than 50 DEG C be concentrated under reduced pressure into it is dry, it is standby.
Step 6：By gains in step 5 through Sephadex LH-20, with methanol-water（70/30, v/v）Isocratic elution, i.e.,
Obtain new skeleton alkaloids compound oleracimine.Through ultra performance liquid chromatography, it is 90-99% that normalization method, which determines purity,.
（2）Purslane source organism alkali is oleracone preparation method, is comprised the following steps.
The step 1-3 in above-mentioned oleracimine preparation methods is repeated, then acetic acid ethyl ester extract in step 3 is done
Method loading, is separated through silica gel column chromatography, and the silica gel is 200-400 mesh, successively with petroleum ether-acetone（1:1,1:2,1:3,1:
5, v:v）Gradient elution obtains 130 elution positions, is designated as eluting position 1-130 in order, is detected through thin-layer chromatography, is shown
Color, merges 90-130 elutions position, dry by being concentrated under reduced pressure into below 40 DEG C of elution position after merging, standby.By in step 4
Gains are separated through ODS medium pressure column chromatographies again, use methanol-water（70/30, v/v）Isocratic elution, obtains 10 elution positions, presses
Order is designated as position 1-10, is detected through thin-layer chromatography, develops the color, and will elute position 3 and is concentrated under reduced pressure into below 50 DEG C dry, and obtain
Concentrate is standby.By in step 5 gained concentrate through Sephadex LH-20, with methanol-water（50/50, v/v）Isocratic elution,
Connect according to color purple band, produce purslane source organism alkali oleracone.Through ultra performance liquid chromatography, normalization method is determined
Purity is 90-99%.
3rd, MTT colorimetric method for determining cell viability, above-mentioned three groups growth period RAW264.7 macrophage inoculations of taking the logarithm respectively
In 96 well culture plates, cell density is 1 × 104Individual/mL, per the μ L of hole 100,37 DEG C of temperature, 5%CO2Under the conditions of after overnight incubation,
Experimental group adds the purslane source organism alkali oleracimine of various concentrations（1-20μM）Or oleracone（1-100μM）,
The LPS for being separately added into final concentration of 1 μ g/mL after 1h to LPS groups and experimental group is incubated, the shadow added after medicine to cell is investigated
Ring.After above-mentioned each group cell culture 24h, the μ L of 5 mg/mL MTT 20,37 DEG C of temperature, 5%CO are added in each hole cell2Condition
It is lower to continue to be incubated after 4h, culture is terminated, liquid in hole is abandoned in suction, and 100 μ L dimethyl sulfoxide (DMSO)s are added per hole（DMSO）, 10min is vibrated,
Make to determine each hole light absorption value at intracellular crystallization fully dissolving, the nm wavelength of ELIASA 570, comparative survival rate of cells result is as schemed
Shown in 1A, 1B, wherein "+" indicates LPS, and "-" represents no LPS or alkaloid, and Figure 1A is that oleracimine is induced LPS
The influence schematic diagram of the survival rate of RAW264.7 macrophages, Figure 1B is that oleracone induces RAW264.7 macrophages to LPS
Survival rate influence schematic diagram.
4th, Ge Lisi is utilized（Griess）Method determines NO content, investigates two breeds of horses bitterroot source organism alkali and LPS is induced
Mouse macrophage RAW264.7 NO yields inhibitory action.Containing 10% after mouse macrophage RAW264.7 passages
Cultivated in the sugared cell culture medium DMEM of height of hyclone, experimental group adds the purslane source oleracimine of various concentrations
（1-20μM）Or oleracone（1-50μM）, in 37 DEG C, 5%CO2Under the conditions of be incubated 1h after use LPS（Final concentration of 1 μ g/mL）Lure
Supernatant is collected after leading inflammatory reaction, 24h.Griess methods determine the content of NO in cell supernatant, according to various concentrations dent
The RAW264.7 cells that amaranth source organism alkali is induced LPS discharge NO influence, to reflect NO levels, as shown in Fig. 2A, 2B,
Wherein "+" indicates LPS, and "-" represents no LPS or alkaloid.Fig. 2A is that oleracimine stimulates RAW264.7 thin LPS
Born of the same parents produce NO inhibitory action schematic diagram, and Fig. 2 B are that oleracone stimulates LPS RAW264.7 cells generation NO inhibitory action
Schematic diagram.From Fig. 2A, 2B, two breeds of horses bitterroot source organism alkali oleracimine and oleracone can suppress LPS and lure
The RAW264.7 cells led produce excessive NO.
5th, ELISA method determines inflammatory factor IL-6, TNF-α and inflammatory mediator PGE2：Exponential phase RAW264.7 is huge
Phagocyte is inoculated in 24 well culture plates, and cell density is 1 × 105Individual/mL, per the mL of hole 1,37 DEG C of temperature, 5%CO2Under the conditions of train
Support overnight, experimental group adds purslane source oleracimine（1-20μM）Or oleracone（1-50μM）Cultivate after 1h,
LPS is added per hole（Final concentration of 1 μ g/mL）, 24h is incubated altogether, and every group of processing repeats 3 holes.ELISA method determines two breeds of horses bitterroot and come
IL-6, TNF-α and the PGE of RAW264.7 macrophages secretes after the biology alkali process of source2Content such as Fig. 3 A, 3B, 4A, 4B,
Shown in 5A, 5B, wherein "+" indicates LPS, and "-" represents no LPS or alkaloid.Fig. 3 A are that oleracimine is stimulated LPS
RAW264.7 cells secrete IL-6 influence schematic diagram, and Fig. 3 B are that oleracone stimulates RAW264.7 cells to secrete IL-6 LPS
Influence schematic diagram, Fig. 4 A are that oleracimine stimulates LPS RAW264.7 cell TNF secretions-α influence schematic diagram, Fig. 4 B
RAW264.7 cell TNF secretion-α influence schematic diagram is stimulated LPS for oleracone, and Fig. 5 A are oleracimine to LPS
Stimulate RAW264.7 cells secretion PGE2Influence schematic diagram, Fig. 5 B are that oleracone stimulates RAW264.7 cells to secrete LPS
PGE2Influence schematic diagram.From Fig. 3 A, 3B, 4A, 4B, 5A, 5B, found using ELISA method, the life of two breeds of horses bitterroot source
Alkaloids can suppress IL-6, TNF-α and the PGE of cell secretion2Deng the expression of related inflammation cell factor.
6th, using detected by Western blot（Western bloting）, exponential phase RAW264.7 macrophages are connect
Plant in 24 well culture plates, cell density is 1 × 105Individual/mL, per the mL of hole 1,37 DEG C of temperature, 5%CO2Under the conditions of cultivate, experiment
Group adds oleracimine（Final concentration of 1-20 μM）After cultivate 1h, add LPS（Final concentration of 1 μ g/mL）, cultivate altogether
24h, every group of processing repeats 3 holes, collects the RAW264.7 macrophages after induction, and addition contains PMSF（Final concentration of 1 mM）'s
4 DEG C of precooling cell pyrolysis liquids are blown and beaten repeatedly, are incubated on ice after 30min, 4 DEG C, 10000g centrifugation 5min, take supernatant to carry out BCA eggs
White quantitative, adding 100 DEG C of heating 10min after appropriate sample-loading buffer makes albuminous degeneration, takes 30 μ g albumen samples in loading wells,
Separated, be transferred on pvdf membrane with 10%SDS-PAGE, skimmed milk power room temperature closing 2h；TBST is washed three times, each 10min；Point
4 DEG C of overnight incubations not in corresponding primary antibody and internal reference；TBST is washed 3 times, under the conditions of 37 DEG C, with 1:The horseradish mistake of 2500 dilutions
The secondary antibody of oxide enzyme mark is incubated 45min；TBST is washed 3 times, each 10min；Finally use chemical luminous substrate（ECL）It is aobvious
Shadow.Protein versus level result is as shown in Fig. 6 A, 6B, 6C, and wherein "+" indicates LPS, and "-" represents no LPS or alkaloid.
Fig. 6 A are the influence schematic diagram for the iNOS albumen that oleracimine stimulates LPS the expression of RAW264.7 cells, and Fig. 6 B are
Oleracimine stimulates LPS the influence schematic diagram of the COX-2 albumen of RAW264.7 cells expression, and Fig. 6 C are oleracimine
The protein band figure of RAW264.7 cells expression is stimulated LPS.From Fig. 6 A, 6B, 6C, sent out using detected by Western blot
Existing, purslane source organism alkali oleracimine can suppress the overexpression of iNOS and COX-2 albumen.
7th, using quantitative real-time PCR（Real-time qPCR）, collect each group macrophage after LPS inductions
RAW264.7, extracts total serum IgE, using NanoDrop 2000c micro-spectrophotometers to RNA's using total RNA extraction reagent box
Concentration and purity are detected；Take the RNA of equivalent to carry out reverse transcription, cDNA is synthesized using reverse transcription reagent box, in Applied
On Biosystems Real-time PCR, using target gene and reference gene as sample, equivalent cDNA is taken to be carried out for template real
When quantitative fluorescent PCR react（Final volume is 20 μ L）, reaction condition is：95 DEG C, 30s；95 DEG C, 5s, 60 DEG C, 31s reacts 40
Circulation.After experiment terminates, with 2-ΔΔCtMethod processing data.With respect to mRNA level in-site as shown in Fig. 7 A, 7B, wherein "+" indicates LPS,
"-" represents no LPS or alkaloid.Fig. 7 A are that oleracimine stimulates LPS the iNOS gene shadows of RAW264.7 cells expression
Schematic diagram is rung, Fig. 7 B are that oleracimine stimulates LPS the COX-2 effect gene schematic diagrames of RAW264.7 cells expression.By scheming
7A, 7B are understood, using quantitative real-time PCR, are found with the increasing of purslane source organism alkali oleracimine concentration
Plus, the iNOS and COX-2 of overexpression mRNA are reduced in RAW264.7 cells, and in concentration dependent.
Test primer sequence as follows：
iNOS（forward）5’-GGTGAAGGGACTGAGCTGTT-3’
（reverse）5’-ACGTTCTCCGTTCTCTTGCAG-3’；
COX-2（forward）5’-TGGTGCCCTGGTCTGATGATG-3’；
（reverse）5’-GTGGTAACCGCTCAGGTGTTG-3’；
β-actin（forward）5’-GTGCTATGTTGCTCTAGACTTCG-3’；
（reverse）5’-ATGCCACAGGATTCCATACC-3’.
In summary, what described two purslane source organism alkali oleracimine and oleracone were induced LPS is huge
Phagocyte RAW264.7 propagation is without influence, safety non-toxic；The purslane source organism alkali oleracimine can effectively suppress
Excessive inflammatory cytokine IL-6, TNF-α and inflammatory mediator NO, PGE produced by the macrophage RAW264.7 of LPS inductions2.Institute
State purslane source organism alkali oleracone can suppress in higher concentrations LPS induction macrophage RAW264.7 produced by
Excessive inflammatory cytokine IL-6, TNF-α and inflammatory mediator NO, PGE2.The purslane source organism alkali oleracimine exists
Albumen and gene level, can effectively suppress two synzyme in the RAW264.7 of LPS inductions（INOS and COX-2）Cross scale
Reach, and in concentration dependant.
Result above is proved：Described two purslane source organism alkali oleracimine and oleracone have good
Anti-inflammatory activity, can prepare anti-inflammatory drug, for preventing or treating by IL-6, TNF-α, NO or PGE as lead compound2It is excessive
Caused by inflammation.
Claims (6)
1. purslane source organism alkali or its salt are used as the application for preparing anti-inflammatory drug, it is characterised in that the purslane source
Alkaloid is oleracimine or oleracone；
The structural formula of the oleracimine is：
；
The structural formula of the oleracone is：
。
2. purslane source organism alkali as claimed in claim 1 or its salt are as the application for preparing anti-inflammatory drug, its feature exists
In the compound or its salt containing oleracimine in the anti-inflammatory drug.
3. purslane source organism alkali as claimed in claim 1 or its salt are as the application for preparing anti-inflammatory drug, its feature exists
In the compound or its salt containing oleracone in the anti-inflammatory drug.
4. purslane source organism alkali as claimed in claim 1 or its salt are as the application for preparing anti-inflammatory drug, its feature exists
In anti-inflammatory inflammation caused by the excessive inflammatory cytokine of macrophage release or inflammatory mediator for suppression.
5. purslane source organism alkali as claimed in claim 4 or its salt are as the application for preparing anti-inflammatory drug, its feature exists
In the inflammatory cytokine is IL-6 or TNF-α, and the inflammatory mediator is NO or PGE2。
6. purslane source organism alkali as claimed in claim 5 or its salt are as the application for preparing anti-inflammatory drug, its feature exists
In the anti-inflammatory drug is that inflammatory cytokine over-expresses inhibitor or macrophage produces the inhibitor of excessive NO.
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| CN108084060B (en) * | 2017-12-07 | 2020-04-21 | 辽宁中医药大学 | Alkaloid oleraurea in purslane and extraction and separation method thereof |
| CN109142701A (en) * | 2018-07-28 | 2019-01-04 | 江西师范大学 | A kind of the anti-inflammatory activity detection method and application of Chinese azalea extract |
| CN110237079B (en) * | 2019-07-01 | 2021-05-14 | 山西大学 | Application of radix stemonae alkaloid analogue |
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| CN116173083B (en) * | 2023-03-10 | 2023-12-19 | 西安交通大学 | Extraction process of anti-inflammatory active ingredients of purslane optimized by response surface method |
| CN116730891B (en) * | 2023-06-28 | 2024-03-01 | 辽宁中医药大学 | Two new alkaloid compounds in purslane and extraction and separation method thereof |
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