CN115025113B - New use of Paeonia lactiflora and Glycyrrhiza decoction polysaccharide extract - Google Patents
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Abstract
The invention discloses application of Paeonia lactiflora and Glycyrrhiza uralensis decoction polysaccharide extract in preparing medicines for treating, preventing, relieving and/or alleviating pain-related diseases and/or inflammation-related diseases. The invention also discloses a pharmaceutical composition which comprises an effective amount of Paeonia lactiflora and Glycyrrhiza glabra polysaccharide extract and pharmaceutically acceptable auxiliary materials. The invention can widen the application range of the peony and licorice decoction, and prompt that the effect of polysaccharide components is emphasized during the use and development of the prescription, so that the loss of the drug effect is avoided.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a novel application of a paeonia lactiflora and licorice decoction polysaccharide extract.
Background
Pain is an unpleasant feeling and emotional experience associated with actual or potential tissue damage, a subjective sensation in the human body. The nature of pain is complex and can be divided into the following six general categories depending on the cause of pain generation: inflammatory pain, neuropathic pain, cancer pain, spastic pain, cardiac pain, and the like.
Pain severely affects mental health, resulting in depression, anxiety, loss of pleasure and affecting overall quality of life. The current development of inflammatory pain medications by western medicine has focused mainly on the study of non-opioid medications, but such medications increase the risk of renal and cardiovascular adverse events. The traditional Chinese medicine has mild medicine property, obvious effect and small side effect on human body, so more and more researchers have started to search for medicines suitable for inflammatory pain from the traditional Chinese medicine.
Paeonia radix Glycyrrhizae decoction is taken from Shang Han Lun (treatise on Cold-induced disease) 29, and has the symptoms of typhoid, floating pulse, spontaneous perspiration, frequent urination, vexation, slight aversion to cold, foot spasm … …, and foot spasm after the foot is cured, it is more suitable for Paeonia radix Glycyrrhizae decoction with which the foot stretches. Paeonia lactiflora (four two), licorice (four two, roasted), the two above materials are decocted with three liters of water to obtain one liter of five drugs, and the drugs are taken after being separated into warm drugs. The peony and licorice decoction is used as a classical prescription for spasmolysis and pain relief, has a remarkable effect due to less medicinal taste, and is promoted and used by the traditional Chinese medicine.
Modern pharmacological researches have found that the decoction mainly contains flavonoids, triterpenoid saponins, monoterpenoid glycosides, and other chemical components, and has the effects of relieving spasm, resisting inflammation, relieving pain, relieving cough and asthma, protecting liver, protecting gastric mucosa, regulating immunity, etc. Paeoniflorin, gallic acid, glycyrrhizin, glycyrrhizic acid and the like are main medicinal effect components.
It is clear that the current research on the components of paeonia lactiflora and licorice Shang Yaoxiao is mainly focused on small molecular substances. Therefore, there is a need to make more intensive researches on the peony and licorice decoction so as to develop more anti-inflammatory and analgesic drugs with better effects.
Disclosure of Invention
The invention aims to provide a new application of Paeonia lactiflora and Glycyrrhiza glabra polysaccharide extract aiming at the defects existing in the prior art.
As one aspect of the present invention, the present invention provides the use of paeonia lactiflora and licorice decoction polysaccharide extract in the preparation of a medicament for treating, preventing, alleviating and/or relieving pain-related diseases and/or inflammation-related diseases.
Preferably, the pain-associated disorder is gastrocnemius spasm, intercostal neuralgia, gastrospasm, stomach pain, abdominal pain, sciatica, gynecological inflammatory abdominal pain or dysmenorrhea.
Preferably, the inflammation-related disorder is duodenal ulcer, atrophic gastritis, gastrointestinal neurosis or acute mastitis.
Preferably, the paeonia lactiflora and licorice decoction polysaccharide extract is obtained by the following method:
preparing a peony and licorice soup;
extracting polysaccharide primary extract from the peony and licorice decoction;
purifying the polysaccharide primary extract to obtain purified polysaccharide;
the purified polysaccharide was lyophilized.
Further preferably, the peony and licorice decoction polysaccharide extract is obtained by the following method:
the weight ratio is (1-3): 1, soaking radix Paeoniae and Glycyrrhrizae radix decoction pieces in water, decocting, filtering, and concentrating to obtain radix Paeoniae and Glycyrrhrizae radix decoction;
separating and purifying the paeonia lactiflora and licorice decoction by using ethanol, and then centrifuging, dissolving the obtained precipitate by using PBS buffer solution to obtain polysaccharide primary extract;
purifying the polysaccharide primary extract by adopting an ion exchange chromatography separation method to obtain purified polysaccharide;
drying the purified polysaccharide.
Preferably, the drying is reduced pressure drying, freeze drying or forced air drying.
Preferably, the Paeonia lactiflora and Glycyrrhiza uralensis decoction polysaccharide extract comprises polysaccharide composed of rhamnose, galactose, glucose and fructose.
As another aspect of the present invention, the present invention also provides a pharmaceutical composition for treating, preventing, alleviating and/or alleviating pain-related diseases and/or inflammation-related diseases, comprising paeonia lactiflora and licorice decoction polysaccharide extract and pharmaceutically acceptable excipients.
Preferably, the pain-associated disorder is gastrocnemius spasm, intercostal neuralgia, gastrospasm, stomach pain, abdominal pain, sciatica, gynecological inflammatory abdominal pain or dysmenorrhea.
Preferably, the inflammation-related disorder is duodenal ulcer, atrophic gastritis, gastrointestinal neurosis or acute mastitis.
Preferably, the paeonia lactiflora and licorice decoction polysaccharide extract is obtained by the following method:
preparing a peony and licorice soup;
extracting polysaccharide primary extract from the peony and licorice decoction;
purifying the polysaccharide primary extract to obtain purified polysaccharide;
drying the purified polysaccharide.
Further preferably, the peony and licorice decoction polysaccharide extract is obtained by the following method:
the weight ratio is (1-3): 1, soaking radix Paeoniae and Glycyrrhrizae radix decoction pieces in water, decocting, filtering, and concentrating to obtain radix Paeoniae and Glycyrrhrizae radix decoction;
separating and purifying the paeonia lactiflora and licorice decoction by using ethanol, and then centrifuging, dissolving the obtained precipitate by using PBS buffer solution to obtain polysaccharide primary extract;
purifying the polysaccharide primary extract by adopting an ion exchange chromatography separation method to obtain purified polysaccharide;
drying the purified polysaccharide.
Preferably, the drying is reduced pressure drying, freeze drying or forced air drying.
As a particularly preferred embodiment, the paeonia lactiflora and licorice decoction polysaccharide extract is obtained by the following method:
(1) Preparation of Paeonia lactiflora and licorice decoction
Weighing radix Paeoniae and Glycyrrhrizae radix decoction pieces according to a weight ratio of 1:1, adding 8 times of water for soaking for 30min, decocting for 40min, filtering with non-woven cloth, decocting residues with 6 times of water for 30min, filtering with non-woven cloth, mixing the two filtrates, and recovering and concentrating to 1g crude drug/mL with a rotary evaporator to obtain radix Paeoniae and Glycyrrhrizae radix decoction;
(2) Crude extraction of polysaccharide in Paeonia lactiflora and Glycyrrhiza decoction
Slowly adding 95% ethanol into radix Paeoniae and Glycyrrhrizae radix decoction, stirring until ethanol saturation is 80%, standing at 4deg.C for 24h, centrifuging at 4000rpm for 20min, separating supernatant and precipitate, and dissolving precipitate with PBS buffer (0.05 mol/L, PH 7.4.7.4) to obtain radix Paeoniae and Glycyrrhrizae radix decoction polysaccharide primary extract;
(3) Separation and purification of polysaccharide by anion exchange chromatography
Taking 100mL of the primary extract of the peony and licorice decoction polysaccharide, and loading the sample onto a chromatographic column. Mobile phase: PBS buffer (0.05 mol/L, PH 7.4); the polysaccharide was eluted with 100% mobile phase A at a flow rate of 1mL/min and the eluted fractions were collected.
(4) Freeze-drying
And freeze-drying the eluted components to obtain freeze-dried powder, namely the Paeonia lactiflora and Glycyrrhiza decoction polysaccharide extract.
Preferably, the Paeonia lactiflora and Glycyrrhiza uralensis decoction polysaccharide extract comprises polysaccharide composed of rhamnose, galactose, glucose and fructose. Wherein, the content of glucose accounts for 30% -35% and the content of fructose accounts for 6% -7%.
Preferably, the pharmaceutically acceptable excipients are any one or more of fillers, disintegrants, lubricants, suspending agents, binders, sweeteners, flavoring agents, preservatives and matrices.
Preferably, the pharmaceutical composition includes, but is not limited to, tablets, hard capsules, soft capsules, granules, pills, powders, oral liquids, inhalants or injections. The pharmaceutically acceptable auxiliary materials comprise: fillers, disintegrants, lubricants, suspending agents, binders, sweeteners, flavoring agents, preservatives, matrices, and the like. The filler comprises: starch, pregelatinized starch, lactose, mannitol, chitin, microcrystalline cellulose, sucrose, and the like; the disintegrating agent comprises: starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, crosslinked polyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, crosslinked sodium carboxymethyl cellulose, and the like; the lubricant comprises: magnesium stearate, sodium lauryl sulfate, talc, silica, and the like; the suspending agent comprises: polyvinylpyrrolidone, microcrystalline cellulose, sucrose, agar, hydroxypropyl methylcellulose, and the like; the binder includes starch slurry, polyvinylpyrrolidone, hydroxypropyl methylcellulose, etc.; the sweetener comprises: saccharin sodium, aspartame, sucrose, sodium cyclamate, glycyrrhetinic acid, etc.; the flavoring agent comprises: sweetener and various flavors; the preservative comprises: nipagin, benzoic acid, sodium benzoate, sorbic acid and salts thereof, benzalkonium bromide, chlorhexidine acetate, eucalyptus oil and the like; the matrix comprises: PEG6000, PEG4000, insect wax, and the like.
As used herein, an "effective amount" or "therapeutically effective amount" refers to a non-toxic, but sufficient amount of a drug or agent that provides the desired effect. In the pharmaceutical compositions of the present invention, an "effective amount" of an ingredient or unit of formulation refers to that amount of the ingredient which provides the desired effect when used in combination with other ingredients. The "effective amount" will vary from subject to subject, depending on the age and general condition of the individual, the particular active agent, and the like. Thus, it is not always possible to refer to an exact "effective amount", however, a suitable "effective amount" in any individual case may be determined by one of ordinary skill in the art using routine experimentation. For example, for a normal adult, administration may be in an effective amount of 20 to 30mg/kg, although this is merely exemplary.
The peony and licorice decoction pieces (or licorice) are all collected in Chinese pharmacopoeia (2015 edition) and accord with various regulations in the pharmacopoeia.
The paeoniflorin and liquorice decoction is taken as a classical prescription with analgesic effect, and the main research object is small molecular compounds represented by paeoniflorin and albiflorin, but the current research does not find the pharmacodynamic effect of macromolecules such as polysaccharide components. The inventor of the invention discovers that the polysaccharide extract of the paeonia lactiflora and licorice decoction has better analgesic and anti-inflammatory effects, the analgesic effect is similar to that of the chemical drug acetaminophen, but the anti-inflammatory capability is better than that of the chemical drug acetaminophen, and the polysaccharide extract of the paeonia lactiflora and licorice decoction is derived from traditional Chinese medicines and has better safety. The invention can widen the application range of the peony and licorice decoction, and prompt that the effect of polysaccharide components is emphasized during the use and development of the prescription, so that the loss of the drug effect is avoided.
Drawings
FIG. 1 is a polysaccharide calibration graph of example 2 of the present invention;
FIG. 2 shows the torsion test result in example 3 of the present invention;
FIG. 3 shows the serum PGE2 levels of mice in example 4 of the present invention (representative and model groups have a very significant difference P < 0.01);
fig. 4 shows that the serum NO levels of mice in example 4 of the present invention (representative and model groups have a very significant difference P < 0.01);
fig. 5 shows the serum IL-10 levels of mice in example 4 of the present invention (representative and model groups have a very significant difference P < 0.01).
Detailed Description
Example 1: preparation of Paeonia lactiflora and Glycyrrhiza decoction polysaccharide extract
(1) Preparation of peony and licorice decoction
Weighing 62g of radix Paeoniae and Glycyrrhrizae radix decoction pieces, soaking in eight times of water for 30min, decocting for 40min, filtering with non-woven cloth, decocting the residues with 6 times of water for 30min, filtering with non-woven cloth, mixing the two filtrates, and recovering and concentrating to 1g crude drug/mL with rotary evaporator to obtain radix Paeoniae and Glycyrrhrizae radix decoction.
(2) Crude extraction of polysaccharide in Paeonia lactiflora and Glycyrrhiza decoction
Slowly adding 95% ethanol into radix Paeoniae and Glycyrrhrizae radix decoction, stirring until ethanol saturation is 80%, standing at 4deg.C for 24h, centrifuging at 4000rpm for 20min, separating supernatant and precipitate, and dissolving precipitate with PBS buffer (0.05 mol/L, PH 7.4.7.4) to obtain radix Paeoniae and Glycyrrhrizae radix decoction polysaccharide primary extract.
(3) Separation and purification of polysaccharide by anion exchange chromatography
Separation was performed using ion exchange chromatography using a packing of 201 x 7 (717) strongly basic anion exchange resin, packed in a volume of about 100mL, as follows:
a, firstly taking a proper amount of resin filler, adding saturated saline water with 3 times of volume, soaking for 18-20h, discarding the saline water, adding clear water, and washing until the discharged water is not yellow.
b, adding 3 times of 2% NaOH solution into the resin filler, soaking for 2 hours, and flushing the resin with clear water until the discharged water is nearly neutral after the alkali solution is discharged.
c, adding 3 times of 5% HCl solution into the resin filler, soaking for 4 hours, discharging acid liquor completely, and washing with water to be neutral.
And d, continuously adding 3 times of 2% NaoH solution into the resin filler, soaking for 4 hours, discharging alkali liquor, washing with clear water to be neutral, and discarding excessive clear water to enable the resin filler to be in a uniform slurry state.
e, taking an empty column, vertically fixing the empty column by using an iron frame table, paving a small cotton block, putting the cotton block into the bottom of the empty column, pressing the cotton block by using a glass rod, closing a chromatographic column piston, and adding a little clear water to infiltrate the cotton block.
And f, continuously stirring the resin filler by using a glass rod, discharging bubbles, slowly pouring the resin into the empty column at a constant speed along the glass rod, and slowly withdrawing the glass rod after pouring so as to avoid generating bubbles.
And g, knocking the column by using a rubber tube, driving away bubbles in the silica gel of the chromatographic column, opening the piston, discharging clear water, allowing the resin to naturally settle, closing the piston when the clear water quickly flows out, and finishing column installation.
The glass rod was drained, 3 volumes of PBS buffer were slowly added to the column, and the flow rate was controlled at 1mL/min in the equilibrated column.
Dissolving the precipitate of radix Paeoniae and Glycyrrhrizae radix decoction with PBS buffer solution, loading onto chromatographic column, and collecting the components flowing out during the loading process to obtain polysaccharide component enrichment.
(4) The effluent is separately lyophilized for further use.
Example 2: detection of polysaccharide component in peony and licorice decoction
(1) Drawing of a Standard Curve
Accurately weighing 2.5mg of anhydrous glucose, placing in a 25mL volumetric flask, and fixing the volume with distilled water to obtain 0.1mg/mL reference substance solution. Accurately sucking 0,0.1,0.2,0.4,0.6,0.8 and 1mL of reference substance solution into a test tube, adding distilled water to complement to 1mL, respectively adding 1mL of 4% phenol solution, uniformly mixing, rapidly adding 7mL of sulfuric acid, shaking uniformly, carrying out water bath at 40 ℃ for 30min, carrying out ice water bath for 5min, measuring the absorbance of the reference substances with different concentrations at the wavelength of 490nm by using an ultraviolet spectrophotometry, and drawing a standard curve by taking the absorbance as an ordinate and the glucose concentration as an abscissa.
(2) Content determination of polysaccharide component in Paeonia lactiflora and Glycyrrhiza decoction
Taking a solution without a reference substance as a blank, respectively taking a radix paeoniae alba decoction and polysaccharide concentrate, diluting to a proper concentration, taking 1mL of a sample, adding 1mL of a 4% phenol solution, uniformly mixing, rapidly adding 7mL of sulfuric acid, shaking uniformly, carrying out water bath at 40 ℃ for 30min, carrying out ice water bath for 5min, measuring absorbance at a wavelength of 490nm by using an ultraviolet spectrophotometry, and calculating the polysaccharide content of each sample according to a standard curve.
(3) Analysis of polysaccharide component in decoction
The composition of rhamnose, galactose, glucose and fructose in the polysaccharide component of the decoction was analyzed using ion chromatography electrochemical methods. Precisely sucking radix Paeoniae and Glycyrrhrizae radix decoction, adding 2mol/L trifluoroacetic acid solution, shaking, sealing, hydrolyzing at 80deg.C for 4 hr, cooling to room temperature, diluting with distilled water, removing hydrophobic compound with RP column (40 μm), analyzing with anion exchange chromatographic column, and eluting with eluent: gradient elution with NaOH/NaOAC, flow rate: 0.45mL/min, column temperature: 30 ℃, sample injection volume: 10 μl, detector: electrochemical detector, reference electrode: pH-Ag/AgCl (PN: 061879, SN: 825604), working electrode: gold electrode (PN: 061875, SN: 19133).
(4) Detection result of polysaccharide component in peony and licorice decoction
The polysaccharide content of the decoction was determined using phenol-sulfuric acid method. The absorbance value is taken as an ordinate, the concentration of anhydrous glucose is taken as an abscissa, and a standard curve is drawn, as shown in FIG. 1, and a linear regression equation of Y= 10.656X-0.0065 (R 2 =0.9997), it is shown that anhydrous glucose has a good linear relationship in the concentration range of 0-0.08mg/mL, and can be used for determining the total polysaccharide content in a sample.
As a result, the total polysaccharide content in the decoction of 1g crude drug/mL of Paeonia lactiflora and Glycyrrhiza uralensis is 175.11mg/mL, which indicates that the decoction contains a large amount of polysaccharide components. And the polysaccharide content of the freeze-dried product of the paeonia lactiflora and licorice decoction polysaccharide obtained by means of separation, purification, freeze-drying and the like is 905.47mg/g. The method provided by the invention can effectively enrich and purify the Paeonia lactiflora and Glycyrrhiza uralensis decoction polysaccharide.
The contents of rhamnose, galactose, glucose and fructose in the Paeonia lactiflora decoction and Paeonia lactiflora decoction polysaccharide are analyzed and determined by using an ion chromatography electrochemical detection mode, and the results are shown in Table 1 and Table 2, wherein the highest glucose content in 1g crude drug/mL decoction is 97.51mg/mL, and the next fructose (24.37 mg/mL). The polysaccharide lyophilized powder of Paeonia lactiflora and Glycyrrhiza uralensis has the highest glucose content (335.279 μg/mg), followed by fructose (68.333 μg/mg).
TABLE 1 analysis of polysaccharide fraction in crude drug/mL decoction of 1g
| Composition of the components | Content (mg/mL) |
| Rhamnose (rhamnose) | 0.54 |
| Galactose | 0.86 |
| Glucose | 91.51 |
| Fructose | 24.37 |
TABLE 2 analysis of the Components in Paeonia lactiflora and Glycyrrhiza decoction polysaccharides
| Composition of the components | Content (μg/mg) |
| Rhamnose (rhamnose) | 1.103 |
| Galactose | 2.988 |
| Glucose | 335.279 |
| Fructose | 68.333 |
Example 3: effect of Paeonia lactiflora and Glycyrrhiza decoction polysaccharide extract on acute pain
(1) The experimental method comprises the following steps:
the torsion method experiments are referred to acetic acid torsion experiments in Experimental pharmacology methodologies (Main edition of pharmacology methodologies (fourth edition): wei Wei, wu Ximei, li Yuan, press: date of publication by people health press: 2010). 24 Kunming mice are randomly divided into 3 groups, 8 mice in each group are respectively a model group, a Paeonia lactiflora and Glycyrrhiza glabra polysaccharide group and a positive drug group. Mice in the model group were intraperitoneally injected with physiological saline at a dose of 0.1ml/10 g. The freeze-dried powder of Paeonia lactiflora and Glycyrrhiza glabra polysaccharide prepared in example 1 was dissolved in physiological saline to prepare a solution with a concentration of 22.4mg/ml, and the solution was administered by intraperitoneal injection to mice of Paeonia lactiflora and Glycyrrhiza glabra polysaccharide group at a dose of 0.1ml/10 g. Acetaminophen injection (Le Furui) was diluted with physiological saline to prepare a solution having a concentration of 0.5mg/ml, and the solution was administered once by intraperitoneal injection to mice in the positive group at a dose of 0.1ml/10 g.
After mice of the Paeonia lactiflora decoction polysaccharide group and the positive drug group are administrated for 60min, an acute pain model is established by injecting 0.6% acetic acid solution into the abdominal cavity of each mouse of the model group, the Paeonia lactiflora decoction polysaccharide group and the positive drug group at a dose of 0.1ml/10g, recording the time of first torsion (the condition that the mice have abdomen concave, trunk and hind limb stretch and buttock rise is torsion reaction) and the number of times of torsion reaction within 15min after acetic acid injection, and calculating the torsion inhibition rate.
Torque inhibition (%) = (average number of torsions in model group-average number of torsions in administration group)/average number of torsions in model group x 100%.
(2) Torsion experiment results:
the torsion test results are shown in fig. 2 and table 3, and compared with the model group, the torsion times of mice with the Paeonia lactiflora and Glycyrrhiza glabra decoction polysaccharide group are obviously reduced (p is less than 0.01).
Table 3: influence of Paeonia lactiflora and Glycyrrhiza decoction polysaccharide on the number of times of acetic acid twisting experiment (x- + -s)
| Group of | Number of examples | Number of times of twisting body | Torsion body inhibition ratio (%) |
| Model group | 8 | 38.4±16.99 | |
| Positive medicine group | 8 | 12.5±6.99*** | 67.44 |
| Paeonia lactiflora and licorice decoction polysaccharide group | 8 | 15.25±9.146** | 60.29 |
(P < 0.01 for the representative and model group and P < 0.001 for the representative and model group)
As can be seen from fig. 2 and table 3, the Paeonia lactiflora and Glycyrrhiza uralensis decoction polysaccharides can effectively reduce the number of times of twisting, relieve acute pain, and the rate of inhibition of twisting of polysaccharide groups is close to that of positive drugs, which indicates that Paeonia lactiflora and Glycyrrhiza uralensis decoction polysaccharides have similar analgesic effect as acetaminophen, but acetaminophen has certain side effect as chemical drugs, paeonia lactiflora and Glycyrrhiza uralensis decoction polysaccharides are extracted from traditional Chinese medicine decoction, and have better biocompatibility.
Example 4: experimental method for influence of Paeonia lactiflora and Glycyrrhiza uralensis decoction polysaccharide extract on inflammatory factor level of acute pain model (1)
The number of Kunming mice is 32, and the mice are randomly divided into 4 groups, 8 mice in each group are respectively a normal group, a model group, a Paeonia lactiflora and Glycyrrhiza glabra polysaccharide group and a positive drug group. Mice were fasted for 12h before the experiment, and were not watered.
Normal and model groups of mice were intraperitoneally injected with physiological saline at a dose of 0.1ml/10 g. The freeze-dried powder of the Paeonia lactiflora and Glycyrrhiza glabra polysaccharide prepared in example 1 is dissolved in physiological saline to prepare a solution with the concentration of 11.8mg/ml, and the medicine is injected into the abdominal cavity of mice of the Paeonia lactiflora and Glycyrrhiza glabra polysaccharide group according to the dosage of 0.1ml/10 g. Acetaminophen injection (Le Furui) was diluted with physiological saline to prepare a solution having a concentration of 0.5mg/ml, and the drug was intraperitoneally injected into mice in the positive group at a dose of 0.1ml/10 g. The medicine is administered once.
After mice in the Paeonia lactiflora and Glycyrrhiza glabra polysaccharide group and the positive drug group are administrated for 60min, an acute pain model is established by injecting 0.6% acetic acid solution into the abdominal cavity of each mouse in the model group, paeonia lactiflora and Glycyrrhiza glabra polysaccharide group and the positive drug group at a dose of 0.1ml/10g, and physiological saline is injected into the abdominal cavity of the normal group of mice.
After 60min of pain model establishment, 10% chloral hydrate 0.04ml/10g was used for intraperitoneal injection to anesthetize mice, eyeballs were taken for blood, blood samples were allowed to stand at room temperature for 30min, centrifugation was performed at 3000r/min for 15min at 4 ℃, the supernatant was taken in an EP tube, and the obtained serum samples were stored in a biological sample library at-80 ℃.
The levels of inflammatory factors PGE2, NO, IL-10 in serum were detected separately using a prostaglandin E2 (PGE 2) test kit (Nanjing institute of biological engineering), a Nitric Oxide (NO) assay kit (microplate method) (Nanjing institute of biological engineering) and an interleukin-10 (IL-10) test kit (Nanjing institute of biological engineering). The results are shown in fig. 3, 4 and 5.
As can be seen from fig. 3, 4 and 5, the administration of the peony and licorice polysaccharide to mice can effectively reduce the levels of the pro-inflammatory factors PGE2 and NO (p < 0.01) in serum, and can also increase the levels of the anti-inflammatory factors IL-10. Wherein, the Paeonia lactiflora and Glycyrrhiza uralensis decoction polysaccharide has better capability of reducing the level of proinflammatory factors than that of the positive medicine paracetamol.
Claims (6)
1. Use of Paeonia lactiflora and Glycyrrhiza uralensis decoction polysaccharide extract in the preparation of a medicament for the treatment, prevention, alleviation and/or alleviation of pain-related diseases and/or inflammation-related diseases;
wherein the Paeonia lactiflora and Glycyrrhiza uralensis decoction polysaccharide extract is obtained by the following method:
the weight ratio is (1-3): 1, soaking radix Paeoniae and Glycyrrhrizae radix decoction pieces in water, decocting, filtering, and concentrating to obtain radix Paeoniae and Glycyrrhrizae radix decoction;
separating and purifying the paeonia lactiflora and licorice decoction by using ethanol, and then centrifuging, dissolving the obtained precipitate by using PBS buffer solution to obtain polysaccharide primary extract;
purifying the polysaccharide primary extract by adopting an ion exchange chromatography separation method to obtain purified polysaccharide;
drying the purified polysaccharide.
2. The use according to claim 1, wherein the pain-related disorder is gastrocnemius spasm, intercostal neuralgia, gastrospasm, gastralgia, abdominal pain, sciatica, gynecological inflammatory abdominal pain or dysmenorrhea.
3. The use according to claim 1, wherein the inflammation-related disorder is duodenal ulcer, atrophic gastritis, gastrointestinal neurosis or acute mastitis.
4. The pharmaceutical composition is characterized by comprising an effective amount of Paeonia lactiflora and licorice decoction polysaccharide extract and pharmaceutically acceptable auxiliary materials;
wherein the Paeonia lactiflora and Glycyrrhiza uralensis decoction polysaccharide extract is obtained by the following method:
the weight ratio is (1-3): 1, soaking radix Paeoniae and Glycyrrhrizae radix decoction pieces in water, decocting, filtering, and concentrating to obtain radix Paeoniae and Glycyrrhrizae radix decoction;
separating and purifying the paeonia lactiflora and licorice decoction by using ethanol, and then centrifuging, dissolving the obtained precipitate by using PBS buffer solution to obtain polysaccharide primary extract;
purifying the polysaccharide primary extract by adopting an ion exchange chromatography separation method to obtain purified polysaccharide;
drying the purified polysaccharide.
5. The pharmaceutical composition of claim 4, wherein the pharmaceutically acceptable adjuvant is any one or more of a filler, a disintegrant, a lubricant, a suspending agent, a binder, a sweetener, a flavoring agent, a preservative, and a matrix.
6. The pharmaceutical composition of claim 4, wherein the pharmaceutical composition is a tablet, hard capsule, soft capsule, granule, pill, powder, oral liquid, inhalant or injection.
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