CN102670695A - Application of saponins in radix trichosanthin in protecting nerve cells after subarachnoid hemorrhage - Google Patents
Application of saponins in radix trichosanthin in protecting nerve cells after subarachnoid hemorrhage Download PDFInfo
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- CN102670695A CN102670695A CN2012101760440A CN201210176044A CN102670695A CN 102670695 A CN102670695 A CN 102670695A CN 2012101760440 A CN2012101760440 A CN 2012101760440A CN 201210176044 A CN201210176044 A CN 201210176044A CN 102670695 A CN102670695 A CN 102670695A
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Abstract
The invention discloses an application of saponins in radix trichosanthin in protecting nerve cells after subarachnoid hemorrhage. Experiments and results thereof show that when the concentration of saponins in radix trichosanthin ranges from 1 to 10 mg/L, the saponins in radix trichosanthin has the capability of protecting nerve cells from being damaged; and when the concentration of saponins in radix trichosanthin ranges from 15 to 25 mg/L, the saponins in radix trichosanthin has the capability of protecting nerve cells from being damaged and also has the functions of restraining the nerve cells from apoptosis and further preventing the nerve cells from death. With the increasing of the concentration of oral saponins in radix trichosanthin, the saponins in radix trichosanthin has stronger protective effect to the nerve cells, and when the concentration of saponins in radix trichosanthin is 25 mg/L, the score of neurological function has no difference from that of the control group. Therefore, the saponins in radix trichosanthin is an important drug in preventing the nerve cells from apoptosis after subarachnoid hemorrhage and in improving protection to the nerve cells, has great application in protecting the function of the nerve cells, and also has great development prospect in preventing nerve cells from apoptosis, improving the protection capacity for the nerve cells, and treating the subarachnoid hemorrhage.
Description
Technical field
The present invention relates to a kind of purposes of Radix Trichosanthis saponin, particularly relate to a kind of Radix Trichosanthis saponin purposes in the neurocyte behind the protection subarachnoid hemorrhage.
Background technology
Subarachnoid hemorrhage is many to be caused by factors such as long-term hypertension, cerebrovascular malformation, rupture of intracranial aneurysm.Though its sickness rate accounts for about 20～30% of cerebrovascular, its morbidity is rejuvenation trend, and fatality rate and the disability rate first place that occupies cerebrovascular disease.Because normal pressure is compeled cerebral tissue behind the intracranial hemorrhage, and blood circulation is obstructed, patient often shows symptoms such as intracranial hypertension, disturbance of consciousness.The incidence and development mechanism of cerebrovascular disease is complicated, relates to the unusual of multisystem too many levels.And cause the change of cerebral tissue structure behind the neuronal apoptosis, increase the weight of brain injury, finally induce an illness increase the weight of and prognosis of patients relatively poor.Thereby the neuroprotective cell, the apoptosis that suppresses the cerebral tissue neurocyte is one of target spot of subarachnoid hemorrhage treatment.Up to now, when treatment subarachnoid hemorrhage disease, also do not have a kind of specific drug, and clinical effectiveness is not fully up to expectations.And Chinese medicine contains plurality of active ingredients, therefore when treatment, can play a role at multi-level, many target spots, too many levels simultaneously, and toxic and side effects is little.
Radix Trichosanthis is the dry root of cucurbitaceous plant Fructus Trichosanthis, mainly from Fructus Trichosanthis (
Trichosanthes kirilowiiMax) and trichosanthes rosthornii Harms
Trichosanthes rosthorniiHarm), and Fructus Trichosanthis and trichosanthes rosthornii Harms all " Chinese pharmacopoeia is recorded by 2010 editions.But be not limited to southern Fructus Trichosanthis (
T.da miaoshanensisC. Y. Cheng et Yueh), big sub-Fructus Trichosanthis (
T.truncataC. B. Clarke), Fructus Trichosanthis Cucumeroidis (
T. cucumeroides(Ser.) Maxim), Fructus Trichosanthis anguinae (
TrichosanthesAnguinaL), trichosanthes japonica Regel (
Trichosanthes kirilowiiMaxim Var japonica (Miq.) kitam), trichosanthes bracteataVoigy (
Trichosanthes tricuspidataLour.), different strain Fructus Trichosanthis (
Trichosanthes dioicaRoxb.), the phoenix melon (
Trichosanthes integrifolia(Roxb.) kur) etc.Contain multiple materials such as trichosanthin, THFPS, Radix Trichosanthis saponin in the Radix Trichosanthis.To Radix Trichosanthis research comparatively deep be trichosanthin; And to the research of Radix Trichosanthis saponin aspect; Rarely seen to its extraction process, antioxidation in vitro (Chen Ying etc. the extraction of Radix Trichosanthis total saponins and remove the research of DPPH free radical effect. the medicine biotechnology; 2010,17 (5): 397-399; Chen Ying etc. the content analysis of total triterpene saponins in the Radix Trichosanthis. Chinese wild plant resource; 2010; 29 (4): 61-63. Chen Ying etc. the extraction process of the preferred Radix Trichosanthis total saponins of orthogonal test. the time precious traditional Chinese medical science traditional Chinese medicines; 2011,189 (7): the research 1668-1669) and in cerebral ischemia (Chen Ying etc. Radix Trichosanthis saponin and the purposes in preparation treatment ischemic cerebrovascular medicine thereof. application number 201010537400.8).And the pharmaceutical research that is carried out in the process of Radix Trichosanthis saponin neurocyte behind the protection subarachnoid hemorrhage is looked into newly through authoritative institution's retrieval, does not appear in the newspapers as yet both at home and abroad at present.
Summary of the invention
The new purposes that the purpose of this invention is to provide a kind of Radix Trichosanthis saponin, i.e. Radix Trichosanthis saponin purposes in the neurocyte behind the protection subarachnoid hemorrhage.
The present invention provides a kind of Radix Trichosanthis saponin purposes in the neurocyte behind the protection subarachnoid hemorrhage.
According to above-mentioned Radix Trichosanthis saponin purposes in the neurocyte behind the protection subarachnoid hemorrhage, the purposes in the anti-apoptosis of Radix Trichosanthis saponin neurocyte after inducing subarachnoid hemorrhage; During purposes in the anti-apoptosis of said Radix Trichosanthis saponin neurocyte after inducing subarachnoid hemorrhage, the concentration of its Radix Trichosanthis saponin is 15～25 mg/L.
According to above-mentioned Radix Trichosanthis saponin purposes in the neurocyte behind the protection subarachnoid hemorrhage, the purposes of Radix Trichosanthis saponin in the neuroprotective cell; During the purposes of said Radix Trichosanthis saponin in the neuroprotective cell, the concentration of its Radix Trichosanthis saponin is 1～10 mg/L.
The Radix Trichosanthis saponin that the present invention relates to subarachnoid hemorrhage after purposes in the neurocyte protection.Wherein, the neuroprotective cytoplasma membrane is avoided the effect that damages behind subarachnoid hemorrhage, and the concentration that activates the Radix Trichosanthis saponin that the signaling molecule that shields in the neurocyte expresses is 1～10 mg/L.
In addition, the Radix Trichosanthis saponin is induced the purposes in the anti-apoptosis of neurocyte behind the subarachnoid hemorrhage; Wherein, avoid neuronal apoptosis, the concentration that suppresses the Radix Trichosanthis saponin of neurocyte dna fragmentationization is 15～25mg/L.
are in order to understand essence of the present invention better; The purposes of pollen saponin tomorrow during neuronal apoptosis, neuroprotective cytoplasma membrane are avoided damaging behind the antagonism subarachnoid hemorrhage, i.e. Radix Trichosanthis saponin purposes in the neurocyte behind the protection subarachnoid hemorrhage with the pharmacological experiment of Radix Trichosanthis saponin and result below.
The preparation of Radix Trichosanthis saponin: extract the Radix Trichosanthis saponin with conventional method, its concrete operation method is:
(1), take by weighing a certain amount of Radix Trichosanthis, Radix Trichosanthis is pulverized, pulverize the back and cross 20～60 mesh sieves; Obtain Snakegourd Root; In Snakegourd Root, add entry or certain density ethanol (alcoholic acid concentration expressed in percentage by weight is generally 50～95%), water or alcoholic acid addition are 5～20 times of Snakegourd Root weight, add to adopt the ultrasonic extraction device behind entry or the ethanol (frequency of said ultrasonic extraction device is 30 kHz; Power is 400～450W) to carry out supersound extraction; Each extraction time is 10～20min, extracts 3 times, and 3 extracting solution are merged;
(2), step (1) being merged the extracting solution obtain adopts abstraction instrument (adopting Ninson ultrasonic extraction appearance) to extract that (extractant that adopts during extraction is any in n-butyl alcohol, chloroform and the ethyl acetate; The extractant that adopts with merge the extracting solution that obtains between the two the volume ratio of addition be 1～50:1), leave and take the organic solvent phase after the extraction, first (model of macroporous adsorbent resin is D101, D101B, D201, D301 through macroporous adsorbent resin with gained solution; D3520, AB-8, DM130, X-5, XDA-1, XDA-1B; XDA-7, H-20, H-30, H-40, H-60, HPD-100; HPD-300, HPD-600, NK-2, NKA-2, any among NK-9 and the WLD; Said macroporous adsorbent resin and the solution volume ratio between the two is 4～10:1) to carry out separation and purification, then adopts silica gel column chromatography to be further purified that (when said employing silica gel column chromatography was further purified, its silica gel order number was 60～300 orders; Adopt " chloroform-methanol-water " three's mixed liquor to carry out eluting as solvent when said employing silica gel column chromatography is further purified, elution flow rate is 8～12 ml/min; Mixed volume ratio between said " chloroform-methanol-water " three is 50～85 ︰, 14～40 ︰ 1～10); Water elution to the clarification of behind the purification its sample being adopted 8 times of column volumes earlier is (when adopting the water elution of 8 times of column volumes; Its elution flow rate is 1～2 ml/min), (during with the ethanol elution of 6 times of column volumes, its elution flow rate is 2～3ml/min to the ethanol elution of 6 times of column volumes of reuse; Alcoholic acid volumetric concentration is 50～60%), collect ethanol elution;
(3), the ethanol elution of step (2) being collected; (pressure is 0.6Mpa, and baking temperature is 50 ℃, and the rotary speed of Rotary Evaporators is 60rmp to adopt Rotary Evaporators to carry out drying; Rotary evaporation drying time is 3 hours); Obtain paste Radix Trichosanthis saponin after the drying, (vacuum is 0.085Mpa, and baking temperature is 55 ℃ at last paste Radix Trichosanthis saponin to be carried out vacuum drying; Be 3 hours drying time), pulverize, obtain the faint yellow Radix Trichosanthis saponin of powdery subsequent use (purity of gained Radix Trichosanthis saponin is greater than 90%).Adjusting to suitable concn during employing gets final product.
The subarachnoid hemorrhage Preparation of model:Mice be divided at random matched group (
n=6) and experimental group (
n=66).Matched group does not all give the treatment of Radix Trichosanthis saponin before modeling with after the modeling, experimental group is irritated stomach with the Radix Trichosanthis saponin of various dose.At first with the dissolving of frozen hemolysate return to 37 ℃ subsequent use.Fasting before the kunming mice art is freely drunk water.After the anesthesia of 10% chloral hydrate lumbar injection (50ml/kg), calvarium portion skin is cut in the sterilization of skin of head conventional topical, exposes skull and removes the cranium film.7
#Injection needle is in the hole of boring the about 1 mm size of a diameter apart from bregma " ten " word seam 3.5 mm places, right side.50 μ l microsyringes (Shanghai light positive Medical Instruments company limited) are drawn subsequent use hemolysate 50 μ l, behind the vertical inserting needle 5mm of pin hole, slowly are injected into cerebral tissue.Annotating the blood time is controlled in 10 ± 2 min, after annotating blood and finishing, and original position let the acupuncture needle remain at a certain point 10min, No. 4 suturing with thread management skins.Mice revives naturally, free diet (seeing accompanying drawing 1 for details).
Adopt the method for Celluar and Molecular Biology, test as follows: research Radix Trichosanthis saponin is to the neurocyte protection of the inductive subarachnoid hemorrhage mice of hemolysate and the influence of anti-apoptosis:
1, detect neuronal apoptosis with the TUNEL method: after matched group and the dewaxing of different experiments group brain tissue slice xylene, gradient ethanol rehydration.Following at 21 ℃～37 ℃ with protease K digesting 15～30 min.PBS adds 37 ℃ of lucifuges of 50 μ l TUNEL reactant mixtures and hatches 60min after cleaning 2 times.After PBS cleans 3 times, with fluorescence microscope behind the anti-fluorescent quenching mounting fluid-tight sheet.The result finds: Radix Trichosanthis saponin 10～25 mg/L concentration were handled the subarachnoid hemorrhage mice in the time of 5 days, suppressed neuronal apoptosis; The Radix Trichosanthis saponin is compared apoptosis with contrast when 1～10mg/L concentration neurocyte quantity does not have obvious change (seeing accompanying drawing 2 for details).
2, detect LDH with the 2,4 dinitrophenyl hydrazine method and express, to judge the cytoplasma membrane protection situation of Radix Trichosanthis saponin: build up the test kit description operation by Nanjing to neurocyte.Get 0.2% different experiments group and the 35 μ l of control group mice brain tissue homogenate and detect the LDH vigor.Establish standard pipe (titer 0.02ml), the blank pipe of standard (distilled water 0.15 ml) respectively, measure pipe (sample 0.035 ml) and measure blank pipe (sample 0.035 ml and distilled water 0.115 ml), add substrate buffer 0.25 ml.Measure Guan Zhongzai and add nadide 0.05 ml, back 37 ℃ of water-bath 15 min.Add behind the 2,4 dinitrophenyl hydrazine of 0.25 ml 37 ℃ of water-bath 15 min again.Add 0.4 mol/L sodium hydroxide, 2.5 ml room temperatures at last and place that the 440nm photometry absorbs behind 3 min.Unit of activity is U/L.LDH activity in the tissue=(OD measures pipe-OD control tube)/(the blank pipe of OD standard pipe-OD) * C normal concentration/C sample to be tested protein concentration.The Radix Trichosanthis saponin sees table 1 for details to the protection of neurocyte:
Table 1 Radix Trichosanthis saponin is to the protection of neurocyte
Annotate: the treatment of Radix Trichosanthis saponin is 5 days in the table 1,
n=6; X ± s;
* P<0.05,
* P<0.01 compare with matched group.
3, detect neurocyte NF-κ B with the SABC method and express, promote the effect of nerve growth to judge the Radix Trichosanthis saponin: after the tissue slice gradient rehydration, normal goats serum sealing 15min behind 95 ℃ of antigen retrieval 10～30min.Add one anti-37 ℃ hatch behind 2～3h with anti-DAB colour developing behind the 30min, the resinene mounting (seeing accompanying drawing 3 for details) of hatching of biotin labeling two.
4, delayed ischemic neurological deficits index determining: carry out function of nervous system's scoring by the Zausinger six-distribution method, mice is divided into 6 grade scorings.0 minute: can not walking freely; 1 minute: under the state of freely walking about to pathological changes to sideway swivel; 2 minutes: catch the Mus tail, mice to pathological changes to sideway swivel; 3 minutes: the lateral pressure resistance for executing to the pathological changes offside descended; 4 minutes: can not stretch pathological changes offside fore paw, even whole body is to the offside flexing; 5 minutes: the impassivity functional impairment.Radix Trichosanthis saponin treatment back subarachnoid hemorrhage mice function of nervous system appraisal result sees table 2 for details.
Table 2 delayed ischemic neurological deficits index determining
Indicate: the treatment of Radix Trichosanthis saponin is 5 days in the table 2,
n=6; X ± s;
* P<0.05,
* P<0.01 compare with matched group.
5, experimental data statistical procedures: experimental data is represented with mean+SD, warp
tCheck:
P<0.05 expression has notable difference,
P<0.01 utmost point significant difference is arranged.
By above-mentioned experiment and result thereof, can draw as drawing a conclusion:
Radix Trichosanthis saponin has the ability that the neuroprotective cell is avoided damaging when 1～10 mg/L concentration; Have the effect that the neuroprotective cell is avoided damaging and when 15～25 mg/L concentration, remove, can also suppress neuronal apoptosis, and then avoid nerve cell death.Along with the rising of oral Radix Trichosanthis saponin concentration, it is strong more to protecting neuron from acute, function of nervous system's scoring and matched group zero difference when 25mg/L.Therefore; The Radix Trichosanthis saponin is an antagonism neuronal apoptosis behind the subarachnoid hemorrhage; And raising is to the material impact medicine of neurocyte protection; To very big purposes arranged aspect the neuroprotective cell function, through avoiding neuronal apoptosis and improve aspect the protective capability treatment subarachnoid hemorrhage of neurocyte to have very big development prospect.
Four, description of drawings:
Fig. 1 is the cardinal principle BIAO and BEN of preparation mice subarachnoid hemorrhage model;
Among Fig. 1: A is the normal mouse cerebral tissue, and B is subarachnoid hemorrhage mouse brain tissue.
Fig. 2 be under the fluorescence microscope shown in neurocyte TUNEL result;
Among Fig. 2: A is a matched group, and B is the neuronal apoptosis after the treatment of 5mg/L amount Radix Trichosanthis saponin, and C is the neuronal apoptosis after the treatment of 20mg/L amount Radix Trichosanthis saponin.
Fig. 3 is the expression of the NF-of neurocyte shown in SABC κ B;
Among Fig. 3: A is a matched group, and B is a 5mg/L amount Radix Trichosanthis saponin treatment group, and C is a 20mg/L amount Radix Trichosanthis saponin treatment group.
Fig. 4 is the expression of the p53 of neurocyte shown in the SABC;
Among Fig. 4: A is a matched group, and B is a 5mg/L amount Radix Trichosanthis saponin treatment group, and C is a 20mg/L amount Radix Trichosanthis saponin treatment group.
Fig. 5 is the expression of the caspase-3 of neurocyte shown in the SABC;
Among Fig. 5: A is a matched group, and B is a 5mg/L amount Radix Trichosanthis saponin treatment group, and C is a 20mg/L amount Radix Trichosanthis saponin treatment group.
Fig. 6 is the expression of the JNK of neurocyte shown in the SABC;
Among Fig. 6: A is a matched group, and B is a 5mg/L amount Radix Trichosanthis saponin treatment group, and C is a 20mg/L amount Radix Trichosanthis saponin treatment group.
Five, the specific embodiment:
Below in conjunction with embodiment the present invention is described further, but does not limit content of the present invention.
Embodiment 1:
The separating and extracting process of Radix Trichosanthis saponin: extract according to conventional method for distilling, the concrete operations step is:
(1), take by weighing a certain amount of Radix Trichosanthis, Radix Trichosanthis is pulverized, pulverize the back and cross 20～60 mesh sieves; Obtain Snakegourd Root; In Snakegourd Root, add entry or certain density ethanol (alcoholic acid concentration expressed in percentage by weight is generally 50～95%), water or alcoholic acid addition are 5～20 times of Snakegourd Root weight, add to adopt the ultrasonic extraction device behind entry or the ethanol (frequency of said ultrasonic extraction device is 30 kHz; Power is 400～450W) to carry out supersound extraction; Each extraction time is 10～20min, extracts 3 times, and 3 extracting solution are merged;
(2), step (1) being merged the extracting solution obtain adopts abstraction instrument (adopting Ninson ultrasonic extraction appearance) to extract that (extractant that adopts during extraction is any in n-butyl alcohol, chloroform and the ethyl acetate; The extractant that adopts with merge the extracting solution that obtains between the two the volume ratio of addition be 1～50:1), leave and take the organic solvent phase after the extraction, first (model of macroporous adsorbent resin is D101, D101B, D201, D301 through macroporous adsorbent resin with gained solution; D3520, AB-8, DM130, X-5, XDA-1, XDA-1B; XDA-7, H-20, H-30, H-40, H-60, HPD-100; HPD-300, HPD-600, NK-2, NKA-2, any among NK-9 and the WLD; Said macroporous adsorbent resin and the solution volume ratio between the two is 4～10:1) to carry out separation and purification, then adopts silica gel column chromatography to be further purified that (when said employing silica gel column chromatography was further purified, its silica gel order number was 60～300 orders; Adopt " chloroform-methanol-water " three's mixed liquor to carry out eluting as solvent when said employing silica gel column chromatography is further purified, elution flow rate is 8～12 ml/min; Mixed volume ratio between said " chloroform-methanol-water " three is 50～85 ︰, 14～40 ︰ 1～10); Water elution to the clarification of behind the purification its sample being adopted 8 times of column volumes earlier is (when adopting the water elution of 8 times of column volumes; Its elution flow rate is 1～2 ml/min), (during with the ethanol elution of 6 times of column volumes, its elution flow rate is 2～3ml/min to the ethanol elution of 6 times of column volumes of reuse; Alcoholic acid volumetric concentration is 50～60%), collect ethanol elution;
(3), the ethanol elution of step (2) being collected; (pressure is 0.6Mpa, and baking temperature is 50 ℃, and the rotary speed of Rotary Evaporators is 60rmp to adopt Rotary Evaporators to carry out drying; Rotary evaporation drying time is 3 hours); Obtain paste Radix Trichosanthis saponin after the drying, (vacuum is 0.085Mpa, and baking temperature is 55 ℃ at last paste Radix Trichosanthis saponin to be carried out vacuum drying; Be 3 hours drying time), pulverize, obtain the faint yellow Radix Trichosanthis saponin of powdery subsequent use (purity of gained Radix Trichosanthis saponin is greater than 90%).Adjusting to suitable concn during employing gets final product.
The faint yellow Radix Trichosanthis saponin of above-mentioned gained is made into 1mg/L, 2.5mg/L, 5mg/L, 7.5mg/L; 10mg/L, 12.5mg/L, 15mg/L, 17.5mg/L; 20mg/L, 22.5mg/L, the 25mg/L isoconcentration, continuous irrigation stomach treatment subarachnoid hemorrhage mice was drawn materials after 5 days.Can see through experimental result: when the drug level of 1～10 mg/L was handled the subarachnoid hemorrhage mice, the expression of neurocyte NF-κ B raise, and the expression of LDH reduces, thereby the neuroprotective cell is avoided damage; And the increase neuronal apoptosis number with drug level constantly reduces during 15～25mg/L concentration, and 10% neuronal apoptosis is only arranged during 25 mg/L concentration, and the apoptotic cell of matched group this moment is 28%; And then neurocyte played a protective role.
Embodiment 2:
The separating and extracting process of Radix Trichosanthis saponin is identical with embodiment 1.
The effect that the Radix Trichosanthis saponin is expressed subarachnoid hemorrhage mice p53:
The faint yellow Radix Trichosanthis saponin of above-mentioned gained is made into 1mg/L, 2.5mg/L, 5mg/L, 7.5mg/L; 10mg/L, 12.5mg/L, 15mg/L, 17.5mg/L; 20mg/L, 22.5mg/L, the 25mg/L isoconcentration, continuous irrigation stomach treatment subarachnoid hemorrhage mice was drawn materials after 5 days.(Santa cruz sc-6243) detects its expression through the SABC method with the p53 specific antibody.Can see through experimental result: with the expression decreased (seeing accompanying drawing 4 for details) of the increase p53 of Radix Trichosanthis saponin concentration.Quantitative analysis shows that the expression of matched group p53 is 0.052763; The expression of low dose group p53 about 0.039358, is compared there was no significant difference with matched group basically, and being expressed in about 0.021859 of p53 during high dose group compared significant difference with matched group.Radix Trichosanthis saponin high dose group is through the apoptosis of the expression antagonism neurocyte of inhibition p53.
Embodiment 3:
The separating and extracting process of Radix Trichosanthis saponin is identical with embodiment 1.
The effect that the Radix Trichosanthis saponin is expressed subarachnoid hemorrhage mice caspase-3:
The faint yellow Radix Trichosanthis saponin of above-mentioned gained is made into 1mg/L, 2.5mg/L, 5mg/L, 7.5mg/L; 10mg/L, 12.5mg/L, 15mg/L, 17.5mg/L; 20mg/L, 22.5mg/L, the 25mg/L isoconcentration, continuous irrigation stomach treatment subarachnoid hemorrhage mice was drawn materials after 5 days.(Santa cruz sc-7148) detects its expression through the SABC method with the caspase-3 specific antibody.Can see through experimental result: with the expression decreased (Fig. 5) of the increase caspase-3 of Radix Trichosanthis saponin concentration.Quantitative analysis shows that the expression of matched group caspase-3 is 0.0761; The expression of low dose group caspase-3 maintains about 0.0602 basically; Compare significant difference with matched group; And the expression of high dose group caspase-3 maintains about 0.0361 basically, compares significant difference with matched group.Radix Trichosanthis saponin high dose group is through the apoptosis of the expression antagonism neurocyte of inhibition caspase-3.
Embodiment 4:
The separating and extracting process of Radix Trichosanthis saponin is identical with embodiment 1.
The effect that the Radix Trichosanthis saponin is expressed subarachnoid hemorrhage mice JNK
The faint yellow Radix Trichosanthis saponin of above-mentioned gained is made into 1mg/L, 2.5mg/L, 5mg/L, 7.5mg/L; 10mg/L, 12.5mg/L, 15mg/L, 17.5mg/L; 20mg/L, 22.5mg/L, the 25mg/L isoconcentration, continuous irrigation stomach treatment subarachnoid hemorrhage mice was drawn materials after 5 days.(Santa cruz sc-572) detects its expression through the SABC method with the JNK specific antibody.Can see through experimental result: with the expression decreased (seeing accompanying drawing 6 for details) of the increase JNK of Radix Trichosanthis saponin concentration.Quantitative analysis shows that the expression of matched group JNK is 0.03156, and its expression of low dose group maintains about 0.0284 basically, and the expression of opposite high dose group JNK maintains about 0.4018 basically, compares significant difference with matched group.The Radix Trichosanthis saponin shields to neurocyte through suppressing the JNK activity behind the subarachnoid hemorrhage.
Claims (5)
1. the Radix Trichosanthis saponin purposes in the neurocyte behind the protection subarachnoid hemorrhage.
2. Radix Trichosanthis saponin according to claim 1 is the purposes in the neurocyte behind the protection subarachnoid hemorrhage, it is characterized in that: the purposes in the anti-apoptosis of Radix Trichosanthis saponin neurocyte after inducing subarachnoid hemorrhage.
3. Radix Trichosanthis saponin according to claim 2 is the purposes in the neurocyte behind the protection subarachnoid hemorrhage; It is characterized in that: during purposes in the anti-apoptosis of said Radix Trichosanthis saponin neurocyte after inducing subarachnoid hemorrhage, the concentration of its Radix Trichosanthis saponin is 15～25 mg/L.
4. Radix Trichosanthis saponin according to claim 1 is the purposes in the neurocyte behind the protection subarachnoid hemorrhage, it is characterized in that: the purposes of Radix Trichosanthis saponin in the neuroprotective cell.
5. Radix Trichosanthis saponin according to claim 4 is the purposes in the neurocyte behind the protection subarachnoid hemorrhage, it is characterized in that: during the purposes of said Radix Trichosanthis saponin in the neuroprotective cell, the concentration of its Radix Trichosanthis saponin is 1～10 mg/L.
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| CN101983637A (en) * | 2010-11-10 | 2011-03-09 | 河南科技学院 | Radix trichosanthis saponin and application of radix trichosanthis saponin in preparing medicine for treating ischemic cerebrovascular diseases |
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