CN102793804A - Viola yedoensis makino petroleum ether extract and medicinal application thereof - Google Patents
Viola yedoensis makino petroleum ether extract and medicinal application thereof Download PDFInfo
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- CN102793804A CN102793804A CN2011101405016A CN201110140501A CN102793804A CN 102793804 A CN102793804 A CN 102793804A CN 2011101405016 A CN2011101405016 A CN 2011101405016A CN 201110140501 A CN201110140501 A CN 201110140501A CN 102793804 A CN102793804 A CN 102793804A
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Abstract
The invention belongs to the field of Chinese medicines and particularly relates to a viola yedoensis makino petroleum ether extract and application of the viola yedoensis makino petroleum ether extract to preparation of medicines for preventing acute lung injury. Viola yedoensis makino is separated and extracted by using petroleum ether, and the average yield is 3.07 percent. The whole experiments prove that the viola yedoensis makino petroleum ether extract has an obvious therapeutical effect on the accurate lung injury of mice. The capillary permeability is reduced, pulmonary edema is relieved, and local inflammatory response of lung is inhibited by the anti-inflammatory mechanism of the viola yedoensis makino petroleum ether extract, so that the acute lung injury can be relieved. The viola yedoensis makino petroleum ether extract can be used for preparing medicines for preventing acute lung injury caused by bacterial infection, non-bacterial infection and other stimulating factors.
Description
Technical field
The invention belongs to the field of Chinese medicines, relate to Chinese medicine Herba Violae ligroin extraction and the purposes in preparation treatment acute lung injury medicine thereof.
Background technology
(acute lung injury ALI) is acute, dyspnea, refractory hypoxemia and the pulmonary edema that carrying out property increases the weight of that is caused by paathogenic factor inside and outside the various lungs beyond the property of heart source in acute lung injury; Clinical manifestation is that a kind of the damage with pulmonary vascular endothelial cell and alveolar epithelial cells is the common nonspecific immune reaction disease of pathological characters; Be that systemic inflammatorome causes the performance (Zhang Zhuo of systemic function obstacle syndrome in pulmonary; Wan Jingyuan; Luo Fuling. etc. Chinese herbal medicine, 2008,39:1196-1199).The factor that ALI will be brought out in this area is divided into bacteroidal and non-bacterial.Severe infections, septicemia, wound, poisoning, a large amount of blood transfusion, extracorporeal circulation and disseminated inravascular coagulation (DIC) etc. all can cause the generation of ALI, and be wherein common with severe infections, the research report; The ALI case fatality rate in serious stage is up to 40% (McIntyre RC; Pulido EJ, Bensard DD, et al. Crit Care Med; 2000,28:3314-3331).Endotoxin (LPS) is one of classical bacillary inducement of bringing out the ALI model; LPS is easy to discerned by immune defense system after getting in the body; Cause lung tissue's damage through some cell mediums that transmit inflammatory reaction, be in particular in that the vascular permeability and pulmonary of initiations such as overexpression of infiltration, the various kinds of cell factor and the adhesion molecule of inflammatory cells such as mononuclearcell increases and pulmonary edema.But the variation basic confirmation medicine through observing above index is to the therapeutical effect of acute lung injury.
Herba Violae is the dry herb of Violaceae Chinese violet Viola yedoensis Makino; For conventional Chinese medicine (Chinese Pharmacopoeia committee. Pharmacopoeia of People's Republic of China, version was one one in 2005. Beijing: Chemical Industry Press, 2005); It is cold in nature; Mildly bitter flavor has effects such as heat-clearing and toxic substances removing, removing heat from blood detumescence, is used to treat symptoms such as carbuncle furuncle, acute mastitis acute appendicitis, erysipelas swell and ache, venom.Modern pharmacology research shows, Herba Violae has effects such as antiinflammatory, antibacterial, anti HIV-1 virus and adjusting immunity.
The mice ear that has the ethyl acetate extract xylol of bibliographical information Herba Violae water decoction and ethanol extraction to bring out has the obvious suppression effect; Escherichia coli, Salmonella, gold-coloured staphylococci, staphylococcus epidermidis also all there is stronger inhibitory action (Chen Hulan; The soup abundant; Chen Xing. Chengdu University of Traditional Chinese Medicine's journal, 2008,31 (2): 52-53).There are some researches show that through internal and external test, the Herba Violae water decoction can be reduced the effect of experiment mice phagocytosis of macrophages and TNF secretion-α; Show that the Herba Violae water decoction can be through regulation and control mouse macrophage activity and then downward modulation immune function of mice (Li Haitao; Zhao Hong, Gu Dingwei, etc. Shandong journal of Chinese medicine; 2004,23 (10): 617-619; Li Haitao, Yang Lin, Zhang Guangling, etc. Huabei Coal Medical Sciences College's journal, 2004,6 (5): 553-554).But above-mentioned research does not all relate to the therapeutical effect of Herba Violae ligroin extraction to acute lung injury.
Summary of the invention
The purpose of this invention is to provide Chinese medicine Herba Violae ligroin extraction and the purposes in preparation treatment acute lung injury medicine thereof.
It is Herba Violae that Herba Violae ligroin extraction of the present invention derives from Chinese crude drug.Herba Violae is the dry herb of Violaceae Chinese violet Viola yedoensis Makino; For conventional Chinese medicine (Chinese Pharmacopoeia committee. Pharmacopoeia of People's Republic of China; Version was one one in 2005. Beijing: Chemical Industry Press, 2005) and, it is cold in nature; Mildly bitter flavor has effects such as heat-clearing and toxic substances removing, removing heat from blood detumescence.
Herba Violae ligroin extraction of the present invention is through composition measurement, and it comprises composition: dihydroactinidiolide, 3,7,11,15-tetramethyl-2-hexadecene-1-alcohol; 6,10,14-trimethyl-2-pentadecanone, positive 16 carbonic acid, palmitoleic acid ethyl ester; Ethyl palmitate, phytol, 9Z, 12Z, 15Z-jeceric acid; 9,12-octadecadienoic ethyl ester, 9Z, 12Z, 15Z-jeceric acid ethyl ester and Ethyl Stearate.
Herba Violae ligroin extraction of the present invention prepares through following method,
Exsiccant Herba Violae medical material (1000 g) is pulverized, and adds 95% ethanol (2500 ml) heating and refluxing extraction (80 ° of C), 3 times; Each 1-2 h merges 95% ethanol extract, extremely does in 80 ° of C decompression recycling ethanols; Add 500 ml aqueous suspensions dissolving,, extract 3-5 time with petroleum ether (60-90 ° of C); Merge petroleum ether extraction liquid, concentrate remove desolvate ligroin extraction (30.85 g), yield 3.07%.
Herba Violae ligroin extraction of the present invention has carried out animal experiment, respectively with various dose 2 mg/kg, and 4 mg/kg, 8 mg/kg administrations, the result confirms that described Herba Violae ligroin extraction has the experimental acute lung injury effect of stronger treatment.
1) the Herba Violae ligroin extraction is used to treat the acute lung injury test:
(1) experiment mice lung-douching fluid mononuclearcell counting
Lung-douching fluid is centrifugal, and 200 μ l normal saline (NS) are collected lower floor's cell deposition, beats even.After the dilution of 0.05% gentian violet solution, optical microscope is counting mononuclearcell number down.The result shows, compares with model group, and the Herba Violae ligroin extraction of various dose all can significantly reduce the increasing of mice mononuclearcell number (P < 0.001).
(2) experiment mice lung-douching fluid RBC
Lung-douching fluid is centrifugal, and 200 μ l NS collect lower floor's cell deposition, beats even.After the dilution of RBC liquid, optical microscope is the counting RBC number down.The result shows, compares with model group, and the Herba Violae ligroin extraction of various dose all can significantly reduce the increasing of mouse red blood cell number (P < 0.01).
(3) experiment mice lung weight in wet base/dry weight is than measuring
Inferior lobe of right lung places filter paper, treats to weigh after filter paper blots surface moisture, as weight in wet base; Be placed on 37 oC incubators oven dry (> 72 h afterwards), weigh up to twice, weight no longer changes, claim dry weight; Calculate lung tissue weight in wet base/dry weight than (being W/D).The result shows, compares with model group, and the Herba Violae ligroin extraction of various dose all can significantly suppress the increasing of mouse lung weight in wet base/dry weight ratio (P < 0.01).
4) experiment mice lung-douching fluid total protein is measured
The Coomassie brilliant blue method detects protein content.Bovine serum albumin is as standard protein solution, the production standard curve.The mouse lung irrigating solution is centrifugal, gets supernatant, dilutes with normal saline.After adding testing sample in the coomassie dye liquor, 540 nm places read trap (A) value in automatic ELIASA (Labsystems Dragon), calculate protein level with standard curve.The result shows, compares with model group, and the Herba Violae ligroin extraction of various dose all can significantly suppress the rising (P < 0.001) of mouse lung lung-douching fluid total protein level.
5) experiment mice lung-douching fluid MPO measures
The mouse lung irrigating solution is centrifugal, gets supernatant, detects by MPO test kit description.The result shows, compares with model group, and various dose Herba Violae ligroin extraction all can significantly reduce the increasing of mouse lung irrigating solution MPO (P < 0.01).
6) mensuration of TNF-α, IL-1 β and IL-6 level in the experiment mice lung-douching fluid
The mouse lung irrigating solution is centrifugal, gets supernatant, detects by cytokine test kit description.The result shows, compares with model group, and 8 mg/kg and 4 mg/kg Herba Violae ligroin extractions all can significantly reduce the rising (P < 0.01) of mouse lung irrigating solution TNF-α, IL-1 β and IL-6 level, 2 mg/>kg Herba Violae stones
Oil ether also has obvious inhibitory action (P < 0.01) to mouse lung irrigating solution IL-1 β.
Experimental result of the present invention shows; Described Herba Violae ligroin extraction has therapeutical effect to acute lung injury, described Herba Violae ligroin extraction can significantly reduce lung tissue edema (weight in wet base/dry weight ratio), obviously suppress that albumen oozes out in the lung-douching fluid, reduce lung leukocyte and erythrocyte aggregation quantity, reduce myeloperoxidase (MPO) (MPO) active, suppress tumor necrosis factor (TNF-α), il-1 β (IL-1 β) and the rising of interleukin-6 (IL-6) level.Described Herba Violae ligroin extraction possibly pass through its anti-inflammatory mechanisms; Reduce vascular permeability and pulmonary, alleviate pulmonary edema and ooze out, suppress pulmonary's local inflammation reaction; Reduce the acute lung injury that multiple stimulating factor causes, further can be made into and prevent and treat serious symptom severe acute respiratory syndrome medicine, the medicine of control bacterial pneumonia medicine or control non-bacterial pneumonia.
Table 1 is the Herba Violae ligroin extraction to each index determining result of chmice acute injury of lung model (X ± SD).
Table 1
Wherein, * and ALI model group compare P < 0.05; * and ALI model group be P < 0.01 relatively; * * and ALI model group be P < 0.001 relatively.
Description of drawings
Fig. 1. ALI mouse lung irrigating solution mononuclearcell count results.
Fig. 2. ALI mouse lung irrigating solution RBC result.
Fig. 3. ALI mouse lung weight in wet base/dry weight is than measuring the result.
Fig. 4. ALI mouse lung total protein is measured the result.
Fig. 5. ALI mouse lung irrigating solution MPO measures the result.
Fig. 6. ALI mouse lung irrigating solution TNF-α measures the result.
Fig. 7. ALI mouse lung irrigating solution IL-1 beta determination result.
Fig. 8. ALI mouse lung irrigating solution IL-6 measures the result.
Fig. 9. control group mice lung pathology slice map.
Figure 10. model group mouse lung pathological section figure.
Figure 11. Herba Violae ligroin extraction 8 mg/kg administration group mouse lung pathological section figure.
Figure 12. Herba Violae ligroin extraction 4 mg/kg administration group mouse lung pathological section figure.
Figure 13. Herba Violae ligroin extraction 2 mg/kg administration group mouse lung pathological section figure.
Figure 14. dexamethasone positive controls 4 mg/kg mouse lung pathological section figure.
The specific embodiment
Exsiccant Herba Violae medical material (1000 g), twist into pieces, add 95% ethanol (2500 ml) heating and refluxing extraction (80 ° of C) 3 times, each 2 h; Merge 95% ethanol extract, to doing, add 500 ml aqueous suspensions dissolving in 80 ° of C decompression recycling ethanols; With petroleum ether (60-90 ° of C) extraction, each 500 ml extract 5 times; Merge petroleum ether extraction liquid, concentrate remove desolvate ligroin extraction (30.85 g), yield is 3.07%.
Table 2 is the yield of Herba Violae ligroin extraction.
Table 2
| |
1 | 2 | Meansigma methods |
| Product output (g) | 33.49 | 28.21 | 30.85 |
| Product yield % | 3.33 | 2.81 | 3.07 |
Adopt Agilent 7890A-5973N type gas chromatography mass spectrometry analyser, the composition in the Herba Violae ligroin extraction that makes is measured, the GC condition is: DB-5 MS quartz capillary chromatographic column (60 m * 250 μ m * 0.25 μ m); Carrier gas is high-purity helium, and flow velocity is 0.8 ml/min, split ratio 20:1; Injector temperature 250 oC, temperature programming, initial temperature is 40 oC; 10 oC/min rise to 260 oC, keep 15 min, sample size 0.4 μ l.The MS condition is: the EI ion source, and electron energy 70 ev, ion source temperature 230 oC, level Four bar temperature 150 oC, sweep limits 35-450 amu is divided into from obtaining 16 peaks; Identified wherein 11, accounted for 95.06% of total content, adopted area normalization method to measure its relative amount, the result shows, the Herba Violae ligroin extraction that the present invention makes; It comprises composition: dihydroactinidiolide, 3,7,11,15-tetramethyl-2-hexadecene-1-alcohol; 6,10,14-trimethyl-2-pentadecanone, positive 16 carbonic acid, palmitoleic acid ethyl ester; Ethyl palmitate, phytol, 9Z, 12Z, 15Z-jeceric acid; 9,12-octadecadienoic ethyl ester, 9Z, 12Z, 15Z-jeceric acid ethyl ester and Ethyl Stearate (as shown in table 3).
The component analysis of table 3 Herba Violae ligroin extraction
48 of BALB/c mouses (19-21 g); Be divided into 6 groups (A, B, C, D, E, F groups) at random by body weight: the A group is model control group; The B group is the ALI model group, and C, D, E group are respectively 8,4 and 2 mg/kg for Herba Violae ligroin extraction various dose group; The F group is the positive group of dexamethasone (4 mg/kg), 8 every group.Except that the A group, other treated animal tracheal instillation LPS 3 mg/kg, time 5-6 min instils.No. 5.0 silk suture mices of three edged needle cervical region skin incision.C, D, E group give 2 h gastric infusions behind the LPS, and dosage is respectively 8,4 and 2 mg/kg; All the other operations of A group are identical with the B group, be with physiologic saline for substitute LPS, as model control group.F group 2 h after giving LPS, tail vein injection gives dexamethasone as positive controls, and dosage is 4 mg/kg.2 h gave 1% CMC (sodium carboxymethyl cellulose), gastric infusion after A, B group gave LPS.Mice is plucked eyeball and gets blood after giving LPS 24 h after the anesthesia.Careful ligation right lung, superior lobe of right lung places 10% formalin; Middle lobe of right lung-80 oC preserves; Inferior lobe of right lung is cut, and the normal saline rinsing is weighed behind the filter paper suck dry moisture.Place the baking oven oven dry no longer to change afterwards, calculate wet dry weight ratio until weight.Left side lung is used the normal saline lavation, and the gained lung-douching fluid is centrifugal, collects supernatant, packing, and-80 oC preserve.Be used to detect indexs such as total protein, myeloperoxidase (MPO) (MPO) and inflammatory factor.
Testing result shows; The Herba Violae ligroin extraction can significantly reduce lung tissue weight in wet base/dry weight ratio, reduce neutrophilic granulocyte and erythrocyte aggregation quantity in the lung-douching fluid, obviously suppresses in the lung-douching fluid albumen and oozes out, reduces that MPO is active, the rising of inhibition tumor necrosis factor (TNF-α), il-1 β (IL-1 β) and interleukin-6 (IL-6) level.The Herba Violae ligroin extraction possibly reduce vascular permeability and pulmonary through its anti-inflammatory mechanisms, alleviates pulmonary edema, improve the pneumonia reaction.
Pathological examination results shows; Compare with model group; The alveolar wall of Herba Violae ligroin extraction various dose group thickens; Inflammatory phenomena such as inflammatory cell infiltration alleviate, no severe haemorrhage phenomenon, and damage has the tangible therapeutical effect that improves to prompting Herba Violae ligroin extraction to the chmice acute lung pathology.
Claims (5)
1. the Herba Violae ligroin extraction is characterized in that, it comprises composition: dihydroactinidiolide, 3,7,11; 15-tetramethyl-2-hexadecene-1-alcohol, 6,10,14-trimethyl-2-pentadecanone, positive 16 carbonic acid, palmitoleic acid ethyl ester; Ethyl palmitate, phytol, 9Z, 12Z, 15Z-jeceric acid; 9,12-octadecadienoic ethyl ester, 9Z, 12Z, 15Z-jeceric acid ethyl ester and Ethyl Stearate; Through following method preparation,
Exsiccant Herba Violae medical material is pulverized, and adds 95% alcohol heating reflux and extracts 80 ° of C; 3 times, each 1-2 h merges 95% ethanol extract, extremely does in 80 ° of C decompression recycling ethanols; Add the aqueous suspension dissolving, use petroleum ether, 60-90 ° of C; Extract 3-5 time, merge petroleum ether extraction liquid, concentrate remove desolvate ligroin extraction.
2. the described Herba Violae ligroin extraction of claim 1 is treated the purposes in the acute lung injury medicine in preparation.
3. the described Herba Violae ligroin extraction of claim 1 is prevented and treated the purposes in the serious symptom severe acute respiratory syndrome medicine in preparation.
4. the described Herba Violae ligroin extraction of claim 1 is prevented and treated the purposes in the bacterial pneumonia medicine in preparation.
5. the described Herba Violae ligroin extraction of claim 1 is prevented and treated the purposes in the non-bacterial pneumonia medicine in preparation.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103977152A (en) * | 2013-02-08 | 2014-08-13 | 复旦大学 | Use of herba violae extracts in preparation of drugs for treatment of fever |
| CN105878366A (en) * | 2015-08-27 | 2016-08-24 | 中国药科大学 | Extract capable of reducing blood lipid, reducing blood glucose and protecting liver |
| CN112773852A (en) * | 2021-03-16 | 2021-05-11 | 宁夏医科大学 | Preparation process of Chinese herbal medicine composition with cell protection effect |
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| CN101648935A (en) * | 2009-09-02 | 2010-02-17 | 董爱文 | Method for extracting apigenin from Chinese violet |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103977152A (en) * | 2013-02-08 | 2014-08-13 | 复旦大学 | Use of herba violae extracts in preparation of drugs for treatment of fever |
| CN105878366A (en) * | 2015-08-27 | 2016-08-24 | 中国药科大学 | Extract capable of reducing blood lipid, reducing blood glucose and protecting liver |
| CN112773852A (en) * | 2021-03-16 | 2021-05-11 | 宁夏医科大学 | Preparation process of Chinese herbal medicine composition with cell protection effect |
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Application publication date: 20121128 |