CN109748981A - A kind of alkali carries method and its application of pachymaran - Google Patents
A kind of alkali carries method and its application of pachymaran Download PDFInfo
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Abstract
The invention belongs to polysaccharide preparation and its applications.The application specifically discloses a kind of alkali carries method of pachymaran, comprising steps of (1) poria cocos sclerotium crushes the 3-10 times of dehydrated alcohol that weight is added, refluxing extraction collects alcohol extracting filter residue；(2) the distilled water refluxing extraction of 3-10 times of its weight is added in filter residue, collects water and mentions filter residue；(3) water mentions the NaOH aqueous solution that 10-60 times of 0.75mol/L-2mol/L of its weight is added in filter residue, and alkali carries filtrate is collected in room temperature extraction；(4) HCl is added in alkali carries filtrate and is neutralized to PH 6-7, must precipitate.It is treating and preventing metabolic syndrome, hyperglycemia, hyperlipidemia, nonalcoholic fatty liver, alcoholic fatty liver, weight-reducing and is adjusting the purposes in intestinal flora drug and health food.
Description
Technical field
The present invention relates to technical field of polysaccharide extraction more particularly to the alkali carries technical fields of pachymaran.
Background technique
Metabolic syndrome (MS) is clinical state of a variety of metabolic disorders generations in same individual, these metabolic disorders include
Obesity, insulin resistance, blood fat disorder, impaired glucose tolerance, hypertension etc..The disease incidence of metabolic syndrome is year by year in recent years
It increases, there are about 1/4 populations to suffer from metabolic syndrome for the International Diabetes Federation estimation whole world in 2005, and latest survey is as the result is shown
The disease incidence of China metabolic syndrome patient 13.8% rose to 33.9% by 2000, number of patients up to 4.54 hundred million people,
This is mainly related with the rapid development of urbanization, industrialization, aging of population and economy.Metabolic syndrome be diabetes B and
The high risk factor of the chronic diseases such as cardiovascular and cerebrovascular diseases, and these chronic diseases seriously endanger the life and health of the mankind, to patient
And its family causes great psychological burden and financial burden.Therefore, the high incidence of metabolic syndrome has caused the world to defend
The attention of raw tissue, prevention and treatment are very urgent.
Poria cocos is the dry sclerotia of polyporaceae fungus Poria cocos (Schw.) Wolf, is that China is common
Traditional Chinese medicine, its property is sweet, light, flat, is included into taste kidney channel, there is the effect of clearing damp and promoting diuresis, beneficial spleen calming heart.Cure mainly the deficiency of vital energy it is a tax on one's mind,
The diseases such as oedema, phlegm retention, vomiting, diarrhea, heat gonorrhea, spermatorrhea, palpitation with fear, amnesia.Han dynasty name is recorded in " golden plaque outline is selected to read " cures Zhang Zhong
Scape creates forties prescriptions such as " Poria cocos Sini Tang ", " wuling powder " based on Poria cocos；" Ce Buming hospital opinion " summarizes
Traditional tcm prescription essence more than 200 since Chinese Tang, wherein have a Poria cocos just accounts for 1/5th.There is statistics to show: 158
In a tcm prescription, have Poria cocos just accounts for 80%.Pachymaran and triterpene are its main active constituents.
Summary of the invention
The object of the present invention is to provide a kind of alkali carries method of pachymaran and its in the treatment and prevention of metabolic syndrome
In purposes.
To achieve the goals above, the invention adopts the following technical scheme:
(1) 3-10 times of dehydrated alcohol of its weight of poria cocos sclerotium crushing addition, refluxing extraction 1-4 times, each time 1-4 are small
When, collect filter residue；
(2) the filter residue drying after ethyl alcohol extracts, is added distilled water refluxing extraction 1-4 times by 3-10 times of its weight, every time
Time is 1-4 hours, collects filter residue.
(3) the filter residue drying after water extracts, the NaOH that 0.75mol/L-2mol/L is added by 10-60 times of its weight are water-soluble
Liquid chamber temperature extracts 1-4 hours, collects filtrate.
(4) HCl is added in filtrate and is neutralized to PH 6-7, suction filtration is precipitated, and precipitating is added 3-5 times and distills water washing 3 times
Above to remove water-soluble impurity, resulting precipitating is lyophilized.
(5) detect that (detector: evaporative light is dissipated to obtained pachymaran using water2659-2424 high performance liquid chromatograph
It penetrates, chromatographic column: TSK G5000PWXL)。
(6) pachymaran obtained by the above method can be used for preparing the drug or food for treating or preventing metabolic syndrome,
Especially in the drug or food of hypoglycemic weight-reducing.
From the point of view of current results of animal, which can effectively reduce HbAle in blood glucose and serum
Protein content, therefore the pachymaran can improve hyperglycemia and good to the long-term control effect of blood glucose.It simultaneously can be significant
Ground reduces the level of total cholesterol, triglyceride, low-density lipoprotein in blood and liver, has adjustment effect to lipid metaboli.
In addition, the pachymaran can significantly reduce the content of glutamic-pyruvic transaminase and glutamic-oxalacetic transaminease in blood, with liver protecting
Effect.Level of inflammation can be reduced by disclosing the polysaccharide to the test result of the inflammatory factor TNF-α in serum.Therefore, the Poria cocos
Polysaccharide can improve a variety of symptoms of metabolic syndrome, including reduce blood glucose, improve lipid metaboli and liver function, reduce level of inflammation
Deng.
Detailed description of the invention
Fig. 1 pachymaran purity detecting HPLC result figure.
Fig. 2 pachymaran hydrolyzes derivatization HPLC result figure.
Fig. 3 pachymaran uv-spectrogram figure.
Fig. 4 pachymaran infared spectrum figure.
Fig. 5 pachymaran13C map figure.
Fig. 6 adipose tissue sections result figure.
Fig. 7 liver tissue slices result figure.
Fig. 8 gives pachymaran rear intestinal flora figure.
Fig. 9 Enterobacter and abundance reduce figure.
Figure 10 Enterobacter and abundance reduce figure.
Specific embodiment
The method and its application that the present invention is further explained in conjunction with specific embodiments.It will be understood by those skilled in the art that
These contents described below are that for a better understanding of the present invention, the scope of the present invention is not limited to this.
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples commercially obtains unless otherwise specified.
Embodiment 1
The extraction of pachymaran:
(1) poria cocos sclerotium 500g is weighed, is crushed, is added 3.5L dehydrated alcohol refluxing extraction 3 times, each time 2 h, is taken out
Filter residue is collected in filter；
(2) the filter residue drying after ethyl alcohol extracts, takes 200g to be added 1L distilled water refluxing extraction 2 times, and each time is 2 small
When, it filters, collects filter residue.
(3) the filter residue drying after water extracts, the NaOH aqueous solution room temperature extraction 3 for taking 5g that 100mL 0.75mol/L is added are small
When, it filters, collects filtrate.
(4) HCl is added in filtrate and is neutralized to PH 6-7, suction filtration is precipitated, and precipitating is added 3-5 times and distills water washing 3 times
Above to remove water-soluble impurity, resulting precipitating is lyophilized, obtains pachymaran 3.45g altogether.
Embodiment 2
The purity detecting and Structural Identification of pachymaran:
Purity detecting: GPC is carried out to 1 gained pachymaran of embodiment using water2659-2424 high performance liquid chromatograph
Detect (detector: evaporative light scattering detector, chromatographic column: TSK G5000PWXL7.8mm × 30cm, 10 μm, mobile phase: water,
Flow velocity: 0.4mL/min, column temperature: 30 DEG C, drift tube temperature: 80 DEG C, atomizer mode: heating (80% power, 48 DEG C), carrier gas:
N2, sampling volume: 10 μ L).Testing result is that single, symmetrical peak proves that the polysaccharide is homogeneous components as shown in Figure 1.
Molecular weight determines: ten illiciumverum degree laser light scattering combined system of SEC-RI-DAWN EOS is utilized, to the pachymaran
Molecular weight is measured.
Determination condition: chromatographic column: SHODEX SB-807HQ；Column temperature: 25 DEG C；Flow velocity: 0.75mL/min；Sampling volume: 10
μL；Sample solution concentration: 4mg/mL；Testing result is as shown in the table:
Monosaccharide composition:
(1) it takes 1 gained pachymaran of 10mg embodiment in ampoule bottle, adds 2mL 2N HCl, seal, 110 DEG C of hydrolysis
Methanol washing is added repeatedly in 180min after cooling, 1mL distilled water is added and makes it dissolve, spare.
(2) using water2659-2424 high performance liquid chromatograph to above-mentioned pachymaran hydrolysate and monosaccharide standard
Detected (detector: evaporative light scattering detector, chromatographic column: Inertsil NH25 μm of 4.6 × 250mm, mobile phase:
Water, flow velocity: 0.6mL/min, column temperature: 30 DEG C, drift tube temperature: 80 DEG C, atomizer mode: heating (80% power, 48 DEG C) carries
Gas: N2, sampling volume: 10 μ L).Testing result hydrolysate as shown in Figure 2 and D-Glucose standard items are completely overlapped, illustrate this
Poria cocos homogeneity polysaccharide is made of D-Glucose.
It is ultraviolet, infrared and13C spectrum information is as in Figure 3-5.
Ultraviolet spectra: 1 gained pachymaran of embodiment is configured to the solution of 2mg/ml with 0.1mol/L sodium hydroxide,
The scanning of 190-400nm wave-length coverage illustrates as a result as shown in figure 3,260nm and 280nm do not absorb without albumen, nucleic acid
Equal impurity.
Infrared spectroscopy: 1 gained pachymaran 5mg of embodiment is weighed, is mixed with dry KBr powder, 5 is ground in mortar
The tabletting on tablet press machine after minute, in 4000~500cm of measurement on infrared spectrometer-1The IR at place.As a result as shown in figure 4,
3395cm-1: hydroxyl；2914cm-1: C-H stretching vibration；890cm-1: β-glycosidic bond.
13C spectrum: 1 gained pachymaran 5mg of Example is dissolved in DMSO-d6Measure its C spectrum, as a result as shown in figure 5, with
The C spectrum of monosaccharide is consistent, thus it is speculated that it is the homopolysaccharide containing a kind of monosaccharide.103.1ppm: for the different head C-1 of β-glucosides.
Embodiment 3
The extraction of pachymaran:
(1) poria cocos sclerotium 500g is weighed, is crushed, is added 3.5L dehydrated alcohol refluxing extraction 3 times, each time 2 h, is taken out
Filter residue is collected in filter；
(2) the filter residue drying after ethyl alcohol extracts, takes 200g to be added 1L distilled water refluxing extraction 2 times, and each time is 2 small
When, it filters, collects filter residue.
(3) the filter residue drying after water extracts, the NaOH aqueous solution room temperature extraction 3 for taking 5g that 100mL 0.5mol/L is added are small
When, it filters, collects filtrate.
(4) HCl is added in filtrate and is neutralized to PH 6-7, suction filtration is precipitated, and precipitating is added 3-5 times and distills water washing 3 times
Above to remove water-soluble impurity, resulting precipitating is lyophilized, obtains pachymaran 0.65g altogether.
Embodiment 4
The extraction of pachymaran:
(1) poria cocos sclerotium 500g is weighed, is crushed, is added 3.5L dehydrated alcohol refluxing extraction 3 times, each time 2 h, is taken out
Filter residue is collected in filter；
(2) the filter residue drying after ethyl alcohol extracts, takes 200g to be added 1L distilled water refluxing extraction 2 times, and each time is 2 small
When, it filters, collects filter residue.
(3) the filter residue drying after water extracts, the NaOH aqueous solution room temperature extraction 3 for taking 5g that 100mL 0.75mol/L is added are small
When, it filters, collects filtrate.
(4) HCl is added in filtrate and is neutralized to PH 6-7, suction filtration is precipitated, and precipitating is added 3-5 times and distills water washing 3 times
Above to remove water-soluble impurity, resulting precipitating is lyophilized, obtains pachymaran 3.33g altogether.
Embodiment 5
The extraction of pachymaran:
(1) poria cocos sclerotium 500g is weighed, is crushed, is added 1.5L dehydrated alcohol refluxing extraction 4 times, 4 hours each time, is taken out
Filter residue is collected in filter；
(2) the filter residue drying after ethyl alcohol extracts, takes 200g to be added 2L distilled water refluxing extraction 4 times, and each time is 4 small
When, it filters, collects filter residue.
(3) the filter residue drying after water extracts, the NaOH aqueous solution room temperature extraction 4 for taking 5g that 300mL 0.8mol/L is added are small
When, it filters, collects filtrate.
(4) HCl is added in filtrate and is neutralized to PH 6-7, suction filtration is precipitated, and precipitating is added 3-5 times and distills water washing 3 times
Above to remove water-soluble impurity, the freeze-drying of resulting precipitating is obtained into pachymaran 3.2g altogether.
Embodiment 6
The extraction of pachymaran:
(1) poria cocos sclerotium 500g is weighed, is crushed, is added 5L dehydrated alcohol refluxing extraction 3 times, 3 hours each time, is taken out
Filter residue is collected in filter；
(2) the filter residue drying after ethyl alcohol extracts, takes 200g to be added 600mL distilled water refluxing extraction 3 times, and each time is 3
Hour, it filters, collects filter residue.
(3) the filter residue drying after water extracts, the NaOH aqueous solution room temperature for taking 5g that 200mL 2mol/L is added extract 2 hours,
It filters, collects filtrate.
(4) HCl is added in filtrate and is neutralized to PH 6-7, suction filtration is precipitated, and precipitating is added 3-5 times and distills water washing 3 times
Above to remove water-soluble impurity, the freeze-drying of resulting precipitating is obtained into pachymaran 3.76g altogether.
Embodiment 7
Pharmacological evaluation is carried out with the pachymaran extract that above-described embodiment 1 is prepared, interpretation of result proves the present invention
Pachymaran extract made from method, which can be applied to preparation, has prevention and treatment metabolic syndrome, hyperglycemia, hyperlipidemia
And the health care product or drug of weight-reducing.
(1) influence of the 1 gained pachymaran of embodiment to ob/ob mouse blood sugar and weight:
Ob/ob mouse (7weeks, ♂, SPF grades) measures fasting blood-glucose and weight, according to blood glucose and weight average grouping point
At 4 groups, every group 10, respectively model group and administration group, ICR mouse (7weeks, ♂, SPF grades) 10 are control group.Grouping
Situation is as follows: (1) blank control group: ICR mouse (gives the distilled water with administration group same dose)；(2) model group: ob/ob
Mouse (gives the distilled water with administration group same dose)；(3) pachymaran high dose group: ob/ob mouse (1g/kg)；(4) Fu
Siberian cocklebur polysaccharide low dose group: ob/ob mouse (500mg/kg)；(5) inulin (positive drug) group: ob/ob mouse (5g/kg).Continuously give
Medicine 5 weeks.
It animal fasting 12 hours after being administered the 35th day, puts to death, heart extracting blood takes liver, kidney and other organs on ice.
Testing index:
1) influence of the administration to blood glucose and weight: the administration same day (0 day), 7 days, 14 days, 21 days, 28 days tail points take blood to survey blood
Sugar value, and weigh and food-intake (being shown in Table 1-3).
2) glucose tolerance test (OGTT): the 30th day survey glucose tolerance of administration: take blood within animal fasting 12 hours (when 0)
Administration, is administered and oral glucose (2.0g/kg), 30,60,120 minutes tail points of difference take blood to survey blood glucose value, draw blood glucose at any time
Between change curve, area under calculated curve.(being shown in Table 4).
3) insulin tolerance tests (ITT): blood (when 0) is taken within 2 hours after administration the 27th day, animal fasting and administration, subcutaneously
Insulin injection (0.4IU/kg B.W., 0.1ml/20g B.W.), takes blood in 40 minutes and 90 minutes after insulin injection
Blood glucose is surveyed, blood glucose is drawn and changes over time curve, area under calculated curve.(table 5)
4) after the last administration, anesthesia is put to death, and takes blood, 4 degrees Celsius of 3000rpm centrifugations, and measurement serum glycated hemoglobin contains
Amount and serum insulin content (being shown in Table 6).
5) after the last administration, anesthesia is put to death, and takes blood, and 4 degrees Celsius of 3000rpm centrifugations measure serum total cholesterol (TC), low
Density lipoprotein (LDL-C), triglycerides (TG), glutamic-pyruvic transaminase (ALT), glutamic-oxalacetic transaminease (AST) (being shown in Table 7).
6) 4) after the last administration, anesthesia is put to death, and takes blood, and 4 degrees Celsius of 3000rpm are centrifuged, measurement Serum TNF-α content (see
Table 8).
7) fatty (Fig. 6) and liver (Fig. 7) after the last administration, are taken, and fixed in formalin, histotomy observation
(Fig. 6-7).
The experimental results showed that compound shown in formula S9 and S24 has remarkable effect to the blood glucose of hyperglycemia mouse, it is negative for sugar
Lotus mouse also has significant decrease to act on (table 14 and table 15).
Influence of 1. pachymaran of table to ob/ob diabetic obese mice food-intake
| Grouping | Total food-intake (g) | Spleen/weight ratio | Kidney/weight ratio |
| (1) | 108.10 | 0.59**** | 1.32**** |
| (2) | 121.172 | 0.22 | 0.66 |
| (3) | 123.59 | 0.32** | 0.72* |
| (4) | 120.72 | 0.33* | 0.70 |
| (5) | 121.69 | 0.27* | 0.63 |
Compared with model group, P < 0.01 * P < 0.05, * *
Influence of 2. pachymaran of table to ob/ob diabetic obese mice weight
Compared with model group, P < 0.01 * P < 0.05, * *
Influence (fasting blood glucose) of 3. pachymaran of table to ob/ob diabetic obese mice blood glucose
Compared with model group, P < 0.01 * P < 0.05, * *
Influence of 4. pachymaran of table to ob/ob diabetic obese mice glucose tolerance
Compared with model group, P < 0.01 * P < 0.05, * *
Influence of 5. pachymaran of table to ob/ob diabetic obese mice insulin tolerance
Compared with model group, P < 0.01 * P < 0.05, * *
Influence of 6. pachymaran of table to ob/ob diabetic obese mice insulin sensitivity index and glycosylated hemoglobin
| Grouping | Insulin sensitivity index | HbA1c/Hb |
| (1) | 0.19** | 0.98* |
| (2) | 0.13 | 1.12 |
| (3) | 0.21** | 0.92** |
| (4) | 0.19* | 0.94* |
| (5) | 0.19** | 0.91** |
Compared with model group, P < 0.01 * P < 0.05, * *
Blood lipid and liver function influence of 7. pachymaran of table to ob/ob diabetic mice
Compared with model group, P < 0.01 * P < 0.05, * *
Influence of 8. pachymaran of table to TNF-α content in ob/ob diabetic obese mice serum
| Grouping | TNF-α content (ng/L) in serum |
| (1) | 401.35* |
| (2) | 530.86 |
| (3) | 443.34* |
| (4) | 475.07 |
| (5) | 470.13 |
From experimental result as can be seen that compared with model group, TNF-α content significantly drops in pachymaran high dose group serum
It is low.
Compared with control group and model group, pachymaran can significantly improve lipid of mice situation, to mice serum ALT,
AST also has good reverse effect, prompts to can protect hepatic injury caused by hyperlipidemia, in addition, pachymaran can significantly drop
The blood glucose of low ob/ob mouse increases insulin sensitivity index, and reduces the content of glycosylated hemoglobin, shows it to ob/ob
The long-term control effect of mouse blood sugar is good, in addition, having a large amount of sizes not in histotomy discovery model group mouse liver cell
Deng fat vacuole, formed fatty liver.And the fat vacuole in pachymaran group mouse liver cell becomes smaller, and prompts pachymaran can
Effectively to treat or alleviate due to diabetes complicated nonalcoholic fatty liver symptom.
(2) influence of the 1 gained pachymaran of embodiment to ob/ob mouse intestinal flora:
1) measurement of intestinal contents 16sDNA
Specification of the extraction of microbial DNA referring to intestinal contents DNA extraction kit.Then Thermo is utilized
The ultraviolet micro-spectrophotometer of NanoDrop 2000 and 1% agargel electrophoresis carry out total DNA quality inspection.Then with dilution after
Genomic DNA is template, carries out PCR using KAPA HiFi Hotstart ReadyMix PCR kit high fidelity enzyme, it is ensured that expand
The Accuracy and high efficiency of increasing.PCR product is detected with 2% agarose gel electrophoresis, and with AxyPrep DNA gel reclaim reagent
Box gel extraction PCR product.After recycling, the ultraviolet micro-spectrophotometer of Thermo NanoDrop 2000 and 2% agar are utilized
Sugared gel electrophoresis carries out library quality inspection.After library quality inspection is qualified, it is quantitative that library is carried out using Qubit, and according to each sample
Data volume requirement, carries out the mixing of corresponding proportion.Using Illumina HiSeq PE250 to the V3- of intestinal flora 16s rDNA
The area V4 carries out classification sequencing.Bioinformatic analysis finally is carried out to sequencing result.
Change of 9. pachymaran of table to ob/ob diabetic obese mice intestinal flora
| Belong to | Model group (Mod) | Pachymaran high dose group (PCP-H) |
| Bacteroides (Bacteroides) | 10.92 | 14.90 |
| Bacillus (Barnesiella) | 16.39 | 11.27 |
| Genus lactubacillus (Lactobacillus) | 10.19 | 6.57 |
| Prey irrigates Pseudomonas (Alloprevotella) | 2.06 | 5.84* |
| Secondary bacteroid (Parabacteroides) | 1.68 | 3.81* |
| Fusobacterium (Clostridium IV) | 1.33 | 3.29 |
| Lachnospira (Lachnospiracea) | 0.13 | 2.88* |
| Sugared bacterium category (Saccharibacteria) | 1.58 | 0.71* |
| Enterobacter (Enterorhabdus) | 1.30 | 0.81 |
| Proteus (Proteus) | 1.21 | 0.01 |
| Ruminococcus (Ruminococcus) | 0.56 | 1.02* |
Compared with model group, P < 0.05 *
PCoA analysis the result shows that, give pachymaran rear intestinal flora has apparent difference (Fig. 8) compared with model group.
Sequencing result shows that the bacterium for sharing 7 doors exists, wherein Bacteroidetes (Bacteroidetes) and Firmicutes
It (Firmicutes) is absolute predominance bacterium in intestinal flora, Firmicutes is rich under report is fat in document and diabetic disease states
Degree increases, and the abundance of Bacteroidetes reduces；Model group mouse Bacteroidetes (Bacteroidetes) and Firmicutes
(Firmicutes) abundance is respectively 58.06% and 28.17%, pachymaran group mouse Bacteroidetes
(Bacteroidetes) and the abundance of Firmicutes (Firmicutes) is respectively 64.34% and 23.38%.
In the level of category, model group mouse gives Lachnospira (Lachnospiracea) after pachymaran, Ruminococcus
Belong to (Ruminococcus), Bacteroides (Bacteroides), Prey irrigates Pseudomonas (Alloprevotella), secondary bacteroid
(Parabacteroides) increase with fusobacterium (Clostridium IV) abundance；Bacillus (Barnesiella), cream
Sour Pseudomonas (Lactobacillus), sugared bacterium category (Saccharibacteria), Enterobacter (Enterorhabdus) and
(Proteus) abundance reduces (Fig. 9-10 and table 9).The Lachnospira wherein dramatically increased after giving Poria cocos
(Lachnospiracea), it is short chain rouge that Ruminococcus (Ruminococcus) and Prey, which irrigate Pseudomonas (Alloprevotella),
Fat acid producing strains, and the Barnesiella and Enterobacter (Enterorhabdus) substantially reduced and fat, insulin resistance
It is formed related.
2) detection of short chain fatty acids (SCFAs)
Stool in mice 30mg after drawing medicine 28 days grinds, sets in 1.5mL centrifuge tube, and 1mL methanol solution, ultrasound is added
It 15 minutes, is centrifuged 5min (5000r/min), supernatant solution is taken to carry out GC-MS analysis.
GC-MS chromatographic condition:
Chromatographic column: Rtx-Wax-60m
Temperature programming: 60 DEG C, 0min is kept；100 DEG C are risen to 5 DEG C/min, keeps 1min；150 are risen to 5 DEG C/min
DEG C, keep 5min；225 DEG C are risen to 30 DEG C/min, keeps 20min.
Carrier gas: He；Flow velocity: 1mL/min；Sampling volume: 2 μ L；It does not shunt.
Mass Spectrometry Conditions: the source EI: mass scan range: 50-800Da, sweep time 10/s；Injection port, transmission line and ion
Source temperature is respectively as follows: 250 DEG C, 250 DEG C and 220 DEG C.
Testing result is as shown in the table:
Influence of 10. pachymaran of table to SCFAs in ob/ob diabetic obese mice excrement
| Acetic acid | Propionic acid | Butyric acid | |
| Model group (Mod) | 1 | 1 | 1 |
| Pachymaran high dose group (PCP-H) | 0.91 | 0.98 | 2.11* |
It is reference with model group, model group SCFAs is set as " 1 ", compared with model group, P < 0.05 *
From experimental result as can be seen that after giving pachymaran, the content of butyric acid is dramatically increased.
In conclusion giving the dynamic equilibrium that pachymaran can significantly adjust intestinal flora, increase probiotics in enteron aisle
Abundance, reduce the abundance of harmful bacteria, increase the generation of butyric acid in enteron aisle.(multiple studies have shown that in SCFAs and glucose body
It is maintained close ties between balance.By some beneficial metabolic effects of propionate and butyric acid Salt treatment by from gut epithelium from the beginning
The glucose of synthesis mediates, sense in portal vein and increased by intestines-cranial nerve loop signal insulin sensitivity and
The such as glucose tolerance)
Influence of 8 embodiment of embodiment, the 1 gained pachymaran to DIO mouse blood sugar and weight:
C57BL/6J (4weeks, ♂, SPF grades) mouse, high lipid food feed 4 weeks, are screened, entered according to body weights
Mouse is selected to be randomly divided into model group and administration group, C57BL/6J mouse (normal diet) is as control.Weight is risen after 4 weeks
20% or more mouse, which is considered as, to be modeled successfully, and fasting blood-glucose is measured, and according to blood glucose value and combines body weights average packet, and every group
10.Grouping situation is as follows: (1) C57BL/6J that blank control group chow diet is fed (gives and administration group same dose
Distilled water)；(2) model group gives the distilled water with administration group same dose；(3) pachymaran high dose group 1g/kg；(4) Fu
Siberian cocklebur polysaccharide low dose group 500mg/kg；(5) inulin (positive drug) organizes 5g/kg.Successive administration 5 weeks.
To weight, the influence of food-intake and blood glucose: administration the 0th day, 7 days, 14 days, 21 days, 28 days, 35 days measurement weight,
Tail point takes the free diet blood glucose of hematometry and fasting blood glucose (after fasting in 4 hours).(table 9-11)
Glucose tolerance test (OGTT): administration the 30th day survey glucose tolerance, take within animal fasting 12 hours blood (when 0) to
Medicine and oral glucose (2.0g/kg), 30,60,120 minutes tail points take blood to survey blood glucose value respectively, draw blood glucose and change over time
Curve, area under calculated curve.(table 12)
Insulin tolerance tests (ITT): taking blood (when 0) for 2 hours after administration the 27th day, animal fasting and administration, subcutaneous to infuse
Insulin (0.4IU/kg B.W., 0.1ml/20g B.W.) is penetrated, takes blood to survey within 40 minutes and 90 minutes after insulin injection
Blood glucose draws blood glucose and changes over time curve, area under calculated curve.(table 13)
Influence of the 9. Poria cocos homogeneous components polysaccharide of table to DIO mouse food-intake
| Grouping | Total food-intake (g) |
| (1) | 106.23 |
| (2) | 124.56 |
| (3) | 123.47 |
| (4) | 125.90 |
| (5) | 122.69 |
Compared with model group, P < 0.01 * P < 0.05, * *
Influence of 10. pachymaran of table to DIO mouse weight
Compared with model group, P < 0.01 * P < 0.05, * *
Influence (fasting blood glucose) of 11. pachymaran of table to DIO mouse blood sugar
Compared with model group, P < 0.01 * P < 0.05, * *
Influence of 12. pachymaran of table to DIO mouse glucose tolerance
Compared with model group, P < 0.01 * P < 0.05, * *
Influence of 13. pachymaran of table to DIO mouse islets element tolerance
Compared with model group, P < 0.01 * P < 0.05, * *
From experimental result as can be seen that compared with model group, the blood glucose and body of DIO mouse is can be significantly reduced in pachymaran
Weight growth rate.It is close with the result of positive drug, but the dosage of above-mentioned pachymaran is only the 1/5 of positive drug inulin dose.
9 embodiment of embodiment, 1 gained pachymaran influences alcoholic liver injury mouse:
C57BL/6J (9weeks, ♂, SPF grades) mouse is randomly divided into three groups, and every group 12, grouping situation is as follows: (1) just
Normal control group Lieber-DeCarli control liquid feed feeds (giving the distilled water with administration group same dose)；(2) model
Group Lieber-DeCarli alcohol liquid forage feed (giving the distilled water with administration group same dose)；(3) pachymaran is low
Dosage group Lieber-DeCarli alcohol liquid forage feed (1g/kg pachymaran).
A weight is weighed every three days, records changes of weight.After being administered the 5th week (the 35th day), on that night that modeling terminates
(last time feeding material) gives the 1/3 of feeding volume according to preceding alcohol liquid Feed consumption for 24 hours, point 3 nursings every time,
It was fed 1 time every 8 hours.
(1) anesthesia is put to death, and takes blood, and AST and ALT (table 14) in blood is surveyed in 4 degrees Celsius of 3000rpm centrifugations.
(2) liver organization is taken, 10% liver tissue homogenate is prepared.Survey the content (table of SOD, malonaldehyde, TG and TC in liver
15-16)。
14. pachymaran of table is to the raised inhibiting effect of alcohol-induced AST, ALT
| Grouping | AST content (IU/L) in serum | Serum alt content (IU/L) |
| (1) | 18.48** | 10.17* |
| (2) | 22.42 | 17.94 |
| (3) | 18.41** | 9.84* |
Compared with model group, P < 0.01 * P < 0.05, * *
15. pachymaran of table is to the raised inhibiting effect of alcohol-induced TG, TC
| Grouping | TG content (mg/g) in liver | TC content (mg/g) in liver |
| (1) | 22.52* | 0.42* |
| (2) | 30.67 | 0.67 |
| (3) | 18.62** | 0.38* |
Compared with model group, P < 0.01 * P < 0.05, * *
Influence of 16. pachymaran of table to alcohol-induced oxidative stress
| Grouping | SOD content (U/mgprot) in liver | Mda content (mmol/mgprot) in liver |
| (1) | 320.77*** | 3.86** |
| (2) | 282.88 | 4.65 |
| (3) | 317.79*** | 3.37* |
Compared with model group, P < 0.001 * P < 0.05, * * P < 0.01, * * *
From experimental result as can be seen that compared with model group, pachymaran can significantly inhibit alcohol-induced mouse blood
The increase of AST in liquid, ALT, and improve alcohol-induced Anomalous lipid metablism, it is solid to reduce triglyceride (TG) and total gallbladder in liver
The content of alcohol (TC), while improving alcohol-induced liver oxidative stress, increase superoxide dismutase from liver (SOD) and contains
Amount, reduces the content of malonaldehyde.In conclusion pachymaran prevention and the treatment alcohol-induced hepatic injury of mouse.
Claims (9)
1. a kind of alkali carries method of pachymaran, includes the following steps:
(1) poria cocos sclerotium crushes the 3-10 times of dehydrated alcohol that weight is added, and refluxing extraction collects alcohol extracting filter residue；
(2) the distilled water refluxing extraction of 3-10 times of its weight is added in filter residue, collects water and mentions filter residue；
(3) water mentions the NaOH aqueous solution that 10-60 times of 0.75mol/L-2mol/L of its weight is added in filter residue, and room temperature extraction is collected
Alkali carries filtrate；
(4) HCl is added in alkali carries filtrate and is neutralized to PH 6-7, must precipitate.
2. purposes of the pachymaran in the drug, health care product, food that preparation treats or prevents metabolic syndrome.
3. pachymaran treats or prevents the purposes in the drug, health care product, food for reducing blood glucose in preparation.
4. purposes of the pachymaran in the drug, health care product, food that preparation treats or prevents hyperlipidemia.
5. purposes of the pachymaran in the drug, health care product, food that preparation treats or prevents nonalcoholic fatty liver.
6. pachymaran treats or prevents the raised drug of TNF-α, health care product, food caused by diabetes or obesity in preparation
In purposes.
7. purposes of the pachymaran in the drug, health care product, food that preparation treats or prevents obesity.
8. purposes of the pachymaran in the drug, health care product, food that preparation adjusts intestinal flora.
9. purposes of the pachymaran in the drug of preparation prevention or treatment alcoholic fatty liver, health care product, food.
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