CN102228517B - Plantain seed extract and application thereof - Google Patents
Plantain seed extract and application thereof Download PDFInfo
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- CN102228517B CN102228517B CN 201110172511 CN201110172511A CN102228517B CN 102228517 B CN102228517 B CN 102228517B CN 201110172511 CN201110172511 CN 201110172511 CN 201110172511 A CN201110172511 A CN 201110172511A CN 102228517 B CN102228517 B CN 102228517B
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Abstract
The invention discloses a Plantain seed extract and application thereof. The Plantain seed extract is acquired by reflux extraction of Plantain seed with 2-6 volume times of 60-90vol% ethanol aqueous solution 2-6 times. The Plantain seed extract contains acteoside, isoacteoside, geniposidic acid, ferulic acid, and Cheqiansu A and Cheqiansu B. In-vivo experiments show that the Plantain seed extract provided by the invention can promote glucose consumption and has lower toxicity to cells; and simultaneously in-vitro experiments show that the Plantain seed extract can significantly increase serum insulin level and reduce fasting blood-glucose level, presents good medicinal value in treatment of diabetes; therefore, the Plantain seed extract can be used in the preparation of pharmaceutical medicaments for treating diabetes singly or together with a pharmaceutically acceptable composition composed of a drug carrier.
Description
Technical field
The present invention relates to a kind of Semen Plantaginis extract and application thereof, belong to technical field of Chinese medicines.
Background technology
Current, diabetes have become one of three large chronic diseases of serious harm modern humans health.The diabetes main manifestations is for having a good appetite, drinking a lot of water, and it is many to urinate, and the symptom of " three-many-one-little " of losing weight simultaneously along with the prolongation of the time of falling ill, can cause the complication of Various Tissues, organ.The medicine that is used for the treatment of at present diabetes mainly contains sulphanylureas, meglitinide, biguanides, thiazolidine dione compounds and alpha-glucosidase inhibitor etc.These medicines have certain therapeutical effect to diabetes, but also are accompanied by simultaneously many side effect, such as gastral untoward reaction and certain Liver and kidney toxicity etc.Simultaneously because diabetes are results that the metabolism of several systems gets muddled in the body, it is the chronic lifelong participation disease of many reasons, patient and government have paid economical and spiritual great cost in the process of long-term treatment, and the new method of therefore seeking the treatment diabetes in Chinese medicine has larger social benefit.
Semen Plantaginis is conventional Chinese medicine, " Chinese pharmacopoeia (2010 editions) is with the dry mature seed of Plantaginaceae Plantago plant Herba Plantaginis Plantago asiatica L. or the Plantago depressa Willd Plantago depressa Willd. Ji Yuan as Semen Plantaginis, has clearing away heat and promoting diuresis, eliminating dampness by diuresis is treating stranguria, the effect such as eliminate the phlegm that makes eye bright is used for the treatment of acute glomerulonephritis, cough with asthma and icterohepatitis etc. traditionally.But Semen Plantaginis also other Chinese medicines of compatibility is used, and for example the compatibility Semen Plantaginis plays the effect of clearing away heat-damp and promoting diuresis in the compound recipe Decoction of Gentiana for Purging the Liver-fire.The alcohol extract of the mutation hair Plantago depressa Willd of discovered in recent years Plantaginaceae plant Plantago depressa Willd (P.depressa var.montata Kitag.) has the effect that certain blood sugar lowering is transferred blood fat to diabetic mice due to the alloxan, its chemical constituent is studied rear discovery, be mainly ursolic acid, oleanolic acid, apigenin, luteolin, catalpol, martynoside, Homoplantaginin, dinatin etc.Other has researcher to find, the methanolic extract of Herba Plantaginis (P.asiatica L.) also has inhibitory action to the formation of advanced glycation end products, demonstrate its potential treating diabetes effect, and to propose glycosylation mortifier main in this extract be Big Semen Plantaginis glycosides (plantamajoside).Simultaneously, the seed shell of Psyllium (P.ovata) has relieving constipation, cholesterol reducing, improves the blood sugar level of diabetics, and has been developed to nutriment.
Summary of the invention
The purpose of this invention is to provide a kind of Semen Plantaginis extract and application thereof.
Described Semen Plantaginis extract is to be that 60～90% ethanol water reflux, extract, obtains for 2～6 times by Semen Plantaginis through the volume fraction of 2～6 times of volumes, contains acteoside, Isoacteoside, Geniposidic acid, ferulic acid, Plantagoamidinic acid A and Plantagoamidinic acid B.
Further, described Semen Plantaginis extract is to be that 70～80% ethanol water reflux, extract, obtains for 3～5 times by Semen Plantaginis through the volume fraction of 3～5 times of volumes.
Further, described Semen Plantaginis extract is to be that 75% ethanol water reflux, extract, obtains for 4 times by Semen Plantaginis through the volume fraction of 4 times of volumes.
Described Semen Plantaginis is recommended as the dry mature seed of Plantaginaceae plant Herba Plantaginis (Plantago asiatica L.) and/or Plantago depressa Willd (Plantago depressa Willd.).
Described Semen Plantaginis recommends to be ground into the granule that diameter is less than or equal to 6mm.
The application of described Semen Plantaginis extract in preparation treatment preparations for diabetes.
The application of the compositions that described Semen Plantaginis extract and pharmaceutically acceptable pharmaceutical carrier form in preparation treatment preparations for diabetes.
Described diabetes are I type and/or type ii diabetes.
Described pharmaceutical dosage forms can be any pharmaceutically useful dosage form, and these dosage forms comprise: tablet, sugar coated tablet, film coated tablet, enteric coated tablet, capsule, hard capsule, soft capsule, oral liquid, suck agent, granule, electuary, pill, powder, unguentum, sublimed preparation, suspensoid, powder, solution, injection, suppository, ointment, plaster, cream, spray, drop, patch; The preferred oral dosage form, as: capsule, tablet, oral liquid, granule, pill, powder, sublimed preparation, unguentum etc.Described peroral dosage form can contain excipient commonly used, such as binding agent, filler, diluent, tablet agent, lubricant, disintegrating agent, coloring agent, flavoring agent and wetting agent, can carry out coating to tablet in case of necessity.Suitable filler comprises cellulose, mannitol, lactose and other similar filler; Suitable disintegrating agent comprises starch, polyvinylpyrrolidone and starch derivatives, for example sodium starch glycollate; Suitable lubricant comprises, for example magnesium stearate.The suitable acceptable wetting agent of medicine comprises sodium lauryl sulphate.
The present invention utilizes the HepG2 cell to carry out the extracorporeal glucose consumption test, the type i diabetes mice that utilizes simultaneously alloxan to cause, the type i diabetes mice that streptozotocin causes, streptozotocin is increased the type ii diabetes rat that the fat feedstuff causes and is carried out effect experiment in the body, the result shows: can promote the consumption of glucose during Semen Plantaginis extract provided by the invention is tested in vivo, and the toxicity to cell is low, simultaneously in vitro tests, can obviously improve serum insulin levels, reduce fasting blood glucose level, demonstrate the medical value of good treatment diabetes (I type and II type).
Description of drawings
Fig. 1 is the finger printing that Semen Plantaginis extract that embodiment 1 obtains obtains under positive ion mode.
Fig. 2 is the finger printing that Semen Plantaginis extract that embodiment 1 obtains obtains under negative ion mode.
Fig. 3 is the mass spectrum that Semen Plantaginis extract that embodiment 1 obtains obtains under positive ion mode.
Fig. 4 is the mass spectrum that Semen Plantaginis extract that embodiment 1 obtains obtains under negative ion mode.
The specific embodiment
Below in conjunction with embodiment the present invention is done further detailed, complete explanation.
Embodiment 1
Remove sandstone and impurity, Semen Plantaginis is pulverized as the granule of 4mm, sieve, with 75% alcohol reflux of 4 times of volumes 4 times, reclaim solvent behind the extracting liquid filtering to dried, use the normal saline suspendible before the use.
It is an amount of to take by weighing the Semen Plantaginis extract sample, (V: V) aqueous solution of formic acid is that the solution that mixes at 90: 10 is dissolved in the volumetric flask with methanol by volume with 0.1%, be diluted to scale, shake up, make every 1mL solution and contain the 1g sample, and utilize Ultra Performance Liquid Chromatography-GC-MS (UPLC-MS) to analyze, analysis condition is as follows:
Chromatographic condition: Waters, Acquity UPLC BEH C
18(50 * 2.1mm) chromatographic columns; (V: the V) aqueous solution of formic acid, Mobile phase B are methanol to mobile phase A mutually in order to contain 0.1% mutually; Flow velocity is 0.3min; Column temperature is 40 ℃; Sample size is 2 μ L; Detecting pattern is the monitoring of negative ions full scan.
The mass spectrum condition: adopt electric spray ion source, when cation full scan detecting pattern carried out data acquisition, ion source temperature was 120 ℃; The desolventizing temperature is 300 ℃; Nitrogen is desolventizing and taper hole gas, and flow velocity is respectively 500L/h and 50L/h; Capillary voltage is 2.2 kilovolts; Taper hole voltage is 23 volts.When anion full scan detecting pattern carried out data acquisition, ion source temperature was 120 ℃; The desolventizing temperature is 300 ℃; Nitrogen is desolventizing and taper hole gas, and flow velocity is respectively 500L/h and 50L/h; Capillary voltage is 2.5 kilovolts; Taper hole voltage is 32 volts.
Fig. 1 is the finger printing that the Semen Plantaginis extract that obtains obtains under positive ion mode, wherein main chromatographic peak is accredited as ferulic acid, Plantagoamidinic acid B, Plantagoamidinic acid A and acteoside, Fig. 2 is the finger printing that the Semen Plantaginis extract that obtains obtains under negative ion mode, and wherein main chromatographic peak is accredited as Geniposidic acid, Plantagoamidinic acid A, acteoside and Isoacteoside; Fig. 3 is the mass spectrum that the Semen Plantaginis extract that obtains obtains under positive ion mode, and Fig. 4 is the mass spectrum that the Semen Plantaginis extract that obtains obtains under negative ion mode; In conjunction with Fig. 1 to Fig. 4 as seen: contain acteoside, Isoacteoside, Geniposidic acid, ferulic acid, Plantagoamidinic acid A and Plantagoamidinic acid B in the Semen Plantaginis extract that obtains.
Embodiment 2
The HepG2 cell is inoculated in 96 orifice plates, treat that cell grows to 60% and merges, change the culture fluid of the variable concentrations Semen Plantaginis extract that contains 0.2% bovine serum albumin (BSA), behind the incubation 24h, detect the glucose residual in the training liquid, be calculated as follows the glucose consumption rate of each porocyte of 24h.
Glucose consumption rate=(experimental port 0h measured value-experimental port 24h measured value)/(control wells 0h measured value-control wells 24h measured value) * 100%.
Table 1 Semen Plantaginis extract is to the facilitation of HepG2 cell glucose metabolism
Annotate: the positive medicine of metformin.
*P＜0.05,
*P＜0.01,
* *P＜0.001 is compared with control wells.
By as seen from Table 1: Semen Plantaginis extract demonstrates the facilitation to the glucose metabolism of HepG2 cell.
Embodiment 3
When the off-test of HepG2 grape cell sugar consumption, culture fluid to be measured is shifted out, change the culture fluid that contains 0.5g/LMTT and continue to hatch 4h, then with culture fluid to the greatest extent, blot, every hole adds the 200l dimethyl sulfoxine again, survey absorbance in the 570nm place on the rearmounted microplate reader of mixing, be calculated as follows the HepG2 cell survival rate: cell survival rate=experimental port measured value/control wells measured value * 100%.
Table 2 Semen Plantaginis extract is to the toxicity of HepG2 cell
Annotate: the positive medicine of metformin.
* *P＜0.001 is compared with control wells.
By as seen from Table 2: with respect to the positive control drug metformin, Semen Plantaginis extract is less to the toxicity of HepG2 cell, demonstrates better safety.
Embodiment 4
Select healthy male mice, (22 ± 2) g is divided into normal group and modeling group at random, modeling group tail vein injection alloxan 48mg/kg.After 3 days, mice fasting 12h, freely drink water, fasting blood glucose level with the determination of glucose oxidase mice, the screening blood glucose value enters experiment greater than the mice of 11.1mmol/L, and be divided into model group and Semen Plantaginis extract group, wherein Semen Plantaginis extract group administration every day Semen Plantaginis extract 300mg/kg.In two weeks of continuous oral gastric infusion, after administration finishes, measure the mice fasting blood sugar.Experimental data is carried out variance analysis, and the result represents with x ± s.
The blood sugar reducing function of the type i diabetes mice that table 3 Semen Plantaginis extract causes alloxan
| Normal group | Model group | The Semen Plantaginis extract group | |
| Fasting blood sugar (mmol/L) | 4.94±0.85 | 19.40±5.12 ### | 10.30±5.10 ** |
Annotate:
###P＜0.001 is compared with normal group;
*P＜0.01 is compared with model group.
By as seen from Table 3: compare with normal group, model group mice fasting blood sugar significantly raises, and type i diabetes model modeling success is described.Simultaneously, compare with model group, the fasting blood sugar of Semen Plantaginis extract group significantly reduces, and illustrates that Semen Plantaginis extract has the effect of significant reduction fasting glucose.
Embodiment 5
Select healthy male mice, (22 ± 2) g is divided into normal group and modeling group at random, and modeling group lumbar injection streptozotocin 150mg/kg cuts tail and surveys random blood sugar behind the 48h, and the mice that the screening blood glucose value surpasses 10.1mmol/L enters experiment.Be Normal group and normal administration group with normal components, the modeling component is model group and model administration group.Normal administration group and model administration group administration every day Semen Plantaginis extract 300mg/kg.In 4 weeks of continuous oral gastric infusion, after administration finishes, measure mice serum insulin and fasting blood sugar.Experimental data is carried out variance analysis, and the result represents with x ± s.
The blood sugar reducing function of the type i diabetes mice that table 4 Semen Plantaginis extract causes streptozotocin
| Normal group | Normal administration group | Model group | Model administration group | |
| Serum insulin value (IU/mL) | 24.56±4.21 | 31.97±7.03 # | 13.50±2.22 ### | 17.21±3.29 * |
| Fasting blood sugar (mmol/L) | 5.31±0.27 | 5.06±0.75 | 29.94±5.18 ### | 23.32±6.62 * |
Annotate:
#P＜0.05,
###P＜0.001 is compared with normal group;
*P＜0.05 is compared with model group.
By as seen from Table 4: compare with Normal group, model group mice fasting blood sugar significantly raises, and type i diabetes model modeling success is described.Simultaneously, compare with model group, Semen Plantaginis extract has significant rising serum insulin and reduces the effect of fasting glucose.Compare with Normal group, normal administration group can make serum insulin raise, and blood glucose is slightly reduced, and the avirulence phenomenon occurs simultaneously, has shown the safety of Semen Plantaginis extract.
Embodiment 6
Select healthy male wistar rat, (200 ± 20) g, be divided at random normal group and high fat group (88% normal diet+10% Adeps Sus domestica+2% cholesterol), feed behind the 3w, high fat group lumbar injection streptozotocin 35mg/kg cuts tail and surveys random blood sugar after five days, the rat that the screening blood glucose value surpasses 16.7mmol/L enters experiment, and be divided into model group and Semen Plantaginis extract group, wherein Semen Plantaginis extract group administration every day Semen Plantaginis extract 150mg/kg.In 4 weeks of continuous oral gastric infusion, after administration finishes, measure rat blood serum insulin and fasting blood sugar.Experimental data is carried out variance analysis, and the result represents with x ± s.
Table 5 Semen Plantaginis extract is to the blood sugar reducing function of type ii diabetes rat
| Normal group | Model group | The Semen Plantaginis extract group | |
| Serum insulin value (IU/mL) | 44.48±12.51 | 17.85±3.78 ### | 25.31±5.52 ** |
| Fasting blood sugar (mmol/L) | 10.12±0.67 | 26.21±1.61 ### | 25.17±5.55 |
Annotate:
###P＜0.001 is compared with normal group;
*P＜0.01 is compared with model group.
By as seen from Table 5: compare with normal group, model group rat fasting blood-glucose value significantly raises, and type ii diabetes model modeling success is described.Simultaneously, compare with model group, Semen Plantaginis extract has significant rising serum insulin and reduces the effect of fasting glucose.
Claims (7)
1. the application of Semen Plantaginis extract in preparation treatment preparations for diabetes, described Semen Plantaginis extract is to be that 60 ~ 90% ethanol water reflux, extract, obtains for 2 ~ 6 times by Semen Plantaginis through the volume fraction of 2 ~ 6 times of volumes, contains acteoside, Isoacteoside, Geniposidic acid, ferulic acid, Plantagoamidinic acid A and Plantagoamidinic acid B.
2. the application of the compositions of a Semen Plantaginis extract and pharmaceutically acceptable pharmaceutical carrier composition in preparation treatment preparations for diabetes, described Semen Plantaginis extract is to be that 60 ~ 90% ethanol water reflux, extract, obtains for 2 ~ 6 times by Semen Plantaginis through the volume fraction of 2 ~ 6 times of volumes, contains acteoside, Isoacteoside, Geniposidic acid, ferulic acid, Plantagoamidinic acid A and Plantagoamidinic acid B.
3. application according to claim 1 and 2 is characterized in that: described Semen Plantaginis extract is to be that 70 ~ 80% ethanol water reflux, extract, obtains for 3 ~ 5 times by Semen Plantaginis through the volume fraction of 3 ~ 5 times of volumes.
4. application according to claim 1 and 2 is characterized in that: described Semen Plantaginis extract is to be that 75% ethanol water reflux, extract, obtains for 4 times by Semen Plantaginis through the volume fraction of 4 times of volumes.
5. application according to claim 1 and 2 is characterized in that: described Semen Plantaginis is the dry mature seed of Plantaginaceae plant Herba Plantaginis and/or Plantago depressa Willd.
6. application according to claim 1 and 2 is characterized in that: described Semen Plantaginis powder is broken into the granule that diameter is less than or equal to 6mm.
7. application according to claim 1 and 2 is characterized in that: described diabetes are I type and/or type ii diabetes.
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| CN102151281B (en) * | 2011-01-24 | 2014-03-26 | 中国药科大学 | Flavonoid compound for preventing and treating diabetes and medicament application thereof |
| CN105796674B (en) * | 2014-12-31 | 2020-08-07 | 上海中医药大学 | Traditional Chinese medicine composition for preventing and treating metabolic syndrome |
| CN106317140B (en) * | 2015-06-29 | 2020-12-04 | 上海中医药大学 | Novel crystal form of geniposide and preparation method and application thereof |
| CN106176777A (en) * | 2016-07-14 | 2016-12-07 | 天津中医药大学 | The new application of rhodioside and be used for treating atherosclerotic compositions |
| CN113384622A (en) * | 2021-07-27 | 2021-09-14 | 上海中医药大学 | Application of semen plantaginis extract in preparing medicine for treating myocardial hypertrophy |
| CN113461749B (en) * | 2021-08-13 | 2023-03-14 | 许昌学院 | Method for extracting geniposide from plantain seeds |
| CN115856108B (en) * | 2022-08-31 | 2024-05-03 | 广东一方制药有限公司 | Construction method and identification method of fingerprint of plantain herb traditional Chinese medicine formula particles |
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| CN102050787A (en) * | 2009-11-09 | 2011-05-11 | 上海中医药大学 | Guanidine derivatives and preparation, medicinal composition and application in preparation of medicaments for treating metabolic syndromes |
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| CN102050787A (en) * | 2009-11-09 | 2011-05-11 | 上海中医药大学 | Guanidine derivatives and preparation, medicinal composition and application in preparation of medicaments for treating metabolic syndromes |
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| 吴斐华等.毛平车前降血糖调血脂作用的研究.《中国中药杂志》.2005,第30卷(第15期),1179-1183. * |
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