CN105520935A - Application of goniolactones C to suppressing proliferation and migration of vascular smooth muscle cells - Google Patents
Application of goniolactones C to suppressing proliferation and migration of vascular smooth muscle cells Download PDFInfo
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Abstract
The invention relates to application of a treatment effective dose of goniolactones C, a medicine composition with the goniolactones C, a medicine box with the goniolactones C and/or an apparatus such as a strut with the goniolactones C and a method for treating or preventing disease states by means of suppressing proliferation and/or migration of smooth muscle cells. Diseases can be selectively blood vessel wall thickening and narrow vessel lumen type cardiovascular and cerebrovascular diseases due to endothelial injury, proliferation of the vascular smooth muscle cells and deposition of matrixes and include diseases due to atherosclerosis, restenosis after angioplasty, hypertension, pulmonary hypertension and diabetic angiopathies.
Description
Technical field
The invention belongs to field of medicaments, be specifically related to brother receive fragrant lactone C and suppress the novelty teabag of vascular smooth muscle cell proliferation and migration, more specifically, the present invention relates to the method suppressing smooth muscle cell proliferation and migration, the administration treatment effective dose taxi driver brother comprised to needs receives fragrant lactone C, and receive the pharmaceutical composition of fragrant lactone C and medicine box containing brother, or treat by suppressing smooth muscle cell proliferation and/or migration or the method for the state that prevents disease, the administration treatment effective dose taxi driver brother comprised to needs receives fragrant lactone C, described disease is selected from because of endothelial injury, vascular smooth muscle cell curing, vessel wall thickening caused by apposition and luminal stenosis cardiovascular disease, comprise atherosclerosis, postangioplasty restenosis, hypertension, pulmonary hypertension, the disease that diabetic angiopathy causes.
Background technology
It is the plant of annonaceae Goniothalamus that Flos Caryophylli brother receives perfume (or spice) (Goniothalamusgriffithii).This platymiscium mostly is the traditional herbal medicine of Southeast Asia and south China provinces and regions, is commonly used for analgesic and parasite killing.At present, in Flos Caryophylli brother receives fragrant plant, extract various active composition, comprising: annonaceous acetogenins (acetogenins), styrylpyrone (styrylpyrones) and alkaloid (alkaloids) compounds.Flos Caryophylli brother receives fragrant extract has stronger cytotoxicity to kinds of tumor cells.
Brother receives fragrant lactone C (GoniolactonesC), be separated to obtain from 95% ethanol extraction of Goniothalamus plant Goniothalamus cheliensis Hu. (GoniothalamuscheliensisHu) rhizome, its structure belongs to styrylpyrone analog, it is as follows that brother receives the structure of fragrant lactone C (GoniolactonesC):
Styrylpyrone has stronger antifungal activity and tumour cell cycle inhibitory action, and on the cell model of diseases of cardiovascular and cerebrovascular systems, rarely have this compounds to report.
Blood vessel wall is made up of tunica intima, middle film and theca externa, is the physiological structure that the critical functions such as the metabolism of tissue and organ, nutrient supply, blood pressure regulating maintain.Tunica intima (tunicaintima) is the innermost layer of tube wall, is made up of endothelium and subendothelial layer, is one deck the thinnest in three layers; Middle film (tunicamedia), between inner membrance and adventitia, forms primarily of smooth muscle cell.In general, blood vessel wall is impaired, the expression of many former tumor genes and cytokine can be caused to generate by intracellular signal transduction, thus the balance of vascular smooth muscle cell proliferation and apoptosis is broken, cause vascular smooth muscle cell to be moved and abnormality proliferation under inner membrance, finally cause vascellum tunica interna incrassation.
Vascellum tunica interna incrassation is cause vessel wall thickening and luminal stenosis cardiovascular and cerebrovascular disease, as the pathologic basis of the disease that atherosclerosis, postangioplasty restenosis, hypertension, pulmonary hypertension, diabetic angiopathy cause.Therefore, by finding anti-smooth muscle cell migration and breeding the medicine reaching and suppress vascellum tunica interna incrassation, alleviate hypertensive vasculopathy, delay end stagerenaldisease, improve diabetic angiopathy, prevention Ischemic Stroke and coronary heart disease, the important means for the treatment of pulmonary hypertension and atherosclerosis disease, and drug development direction.
Research shows, platelet-derived growth factor (PlateletDerivedGrowthFactor, PDGF) is important factor,mitogenic, comprise three kinds of main dimeric structures, i.e. PDGF-AA, PDGF-BB and PDGF-AB, can promote specific cells group division growth.When blood vessel is subject to mechanical damage or other factors stimulate, local PDGF-BB secretion increases.On the latter and smooth muscle cell membrane, β type pdgf receptor (PDGFR) combines, two acceptor molecules are impelled to form dimer, active cell intracellular domain tyrosine residue autophosphorylation, and impel the tyrosine residue phosphorylation of target protein, thus signal is imported in cell, the vital movement of water fall effect regulating cell is amplified through tandem type, comprise the division growth of target cell, therefore blocking PDGF/PDGFR intracellular signaling, is find the important channel with anti-smooth muscle cell proliferation and even anti-angiogenic remodeling activities compound.The present invention receives fragrant lactone C and inhibits vascular remodeling key link by PDGR/PDGFR path having set forth brother first---vascular smooth muscle cell abnormal migration and propagation, and then inhibit blood vessel epimatrix to deposit, therefore have prevention and or treatment vascular remodeling disease lateral reactivity, be that brother receives the novelty teabag of fragrant plant extract GoniolactonesC, there is novelty and novelty.
Summary of the invention
The object of the present invention is to provide brother to receive fragrant lactone C suppresses in the medicine of smooth muscle cell proliferation and/or migration, pharmaceutical composition and pastille material installation application in preparation.
One aspect of the present invention provides brother and receives the application that fragrant lactone C suppresses in preparation in the medicine of smooth muscle cell proliferation and/or migration, pharmaceutical composition and pastille material installation, and the administration comprised to needs is treated effective dose taxi driver brother and received fragrant lactone C.
Further, described smooth muscle cell proliferation and/or migration are the migrations of vascular smooth muscle cell quantity increase and/or the position including but not limited to that the factor such as endothelial injury, exogenous material stimulation causes, and are the pathologic basis causing vessel wall thickening and luminal stenosis cardiovascular and cerebrovascular disease.
Present invention also offers brother and receive the application of fragrant lactone C in preparation prevention or treatment vessel wall thickening and/or the medicine of luminal stenosis cardiovascular and cerebrovascular disease, pharmaceutical composition and pastille material installation.
Further, the disease that described vessel wall thickening and luminal stenosis cardiovascular and cerebrovascular disease are selected from atherosclerosis, postangioplasty restenosis, hypertension, pulmonary hypertension, diabetic angiopathy cause.
Further, described atherosclerosis, be selected from the aortic aneurysm because atherosclerosis of aorta causes, the coronary stricture caused because of coronary atherosclerosis or obstruction (coronary atherosclerotic heart disease), the angiostenosis caused because cranium brain is atherosis, cerebral blood supply insufficiency or local thrombus are formed or plaque rupture, the vascular dementia that fragment comes off in ischemic cerebral apoplexies such as causing cerebral embolism and chronic cerebral ischemia causes, the renal artery thrombosis renal ischemic caused because of atherosclerotic renal artery stenosis the kidney failure finally caused, the acra blood caused because artery of extremity is atherosis also finally develops into intermittent claudication and gangrene for obstacle, and the dyspepsia to cause because of superior mesenteric atherosclerosis, intestinal hypotension, constipation and stomachache.The disease that described diabetic angiopathy causes is selected from diabetic nephropathy, diabetic renal papillary necrosis, diabetic neuropathy, diabetic foot.
The invention still further relates to brother and receive the application that the pharmaceutical preparation of fragrant lactone C suppresses in preparation in the medicine of smooth muscle cell proliferation and/or migration and tube wall thickening and/or luminal stenosis cardiovascular and cerebrovascular disease and pastille material installation.Described pharmaceutical preparation comprises brother and receives fragrant lactone C and pharmaceutically acceptable carrier and/or adjuvant, and described preparation can be the pharmaceutically acceptable pharmaceutical dosage forms such as tablet, capsule, granule, oral liquid, drop pill, micropill, injection.
The described brother of comprising receives the pharmaceutical preparation of fragrant lactone C and method well known in the art can be utilized to prepare.
The dosage of the compounds of this invention pharmaceutical composition is according to preventing or the character of disease therapy and the order of severity, and the individual instances of patient or animal, route of administration and dosage form etc. can have large-scale change.
Beneficial effect of the present invention is: the present invention by PDGF-BB as derivant, observe brother receive smooth muscle cell proliferation that fragrant lactone C induces PDGF-BB, migration, DNA synthesis and cell generation cycle effect and extracellular matrix synthesis and release effects, further illustrate brother and receive smooth muscle cell proliferation that fragrant lactone can suppress PDGF-BB induce, migration and extracellular matrix and synthesize.By to pdgf receptor kinase Activity determination, find that brother receives fragrant lactone C and can directly act on pdgf receptor, suppress its kinase activation, thus the smooth muscle cell proliferation of minimizing PDGF-BB induction, migration and substrate synthesis, the novelty teabag being applicable to suppress vascular smooth muscle cell proliferation and migration, for preventing and/or treating vessel wall thickening because smooth muscle cell proliferation and migration cause and/or luminal stenosis cardiovascular and cerebrovascular disease provides a kind of efficient, safe and economic solution.
Accompanying drawing illustrates:
Fig. 1: brother receives fragrant lactone C and suppresses the VSMC multiplication effect of Serum-induced;
Fig. 2: brother receives the inhibitory action of fragrant lactone C to the VSMC multiplication effect of different growth factor-induced;
Fig. 3: brother receives the inhibitory action that fragrant lactone C synthesizes rat smooth muscle cells DNA;
Fig. 4: brother receive fragrant lactone C suppress PDGF-BB induction VSMC proliferating cycle;
Fig. 5: brother receive fragrant lactone C suppress PDGF-BB induction migration of vascular smooth muscle cells; Fig. 6: brother receive fragrant lactone C suppress PDGF-BB induction adhesion molecule release;
Fig. 7: brother receives fragrant lactone C to the effect of PDGFR kinase activity.
Detailed description of the invention
The following examples are used for further illustrating the present invention, but this and do not mean that any restriction to protection scope of the present invention.Wherein, GoniolactoneC for brother receives fragrant lactone C.
Embodiment 1. brother receives fragrant lactone C and suppresses the proliferation of vascular smooth muscle cells of Serum-induced
Experimental technique: after male SD rat sacrificed by decapitation, takes out thoracic aorta blood vessel rapidly, is separated its fat deposit, is placed in Digestive system (0.06% pancreatin+0.06%I Collagenase Type), in incubator, digests 20-30min under aseptic condition; Take out, remove adventitia completely, with its inner membrance of tweezers tractive angiolysis, smooth muscle layers of making even, shreds, and is placed in culture dish; Cultivate with DMEM/F12 culture medium+20%FBS; 0.125% trypsinization is used when going down to posterity; Culture medium is changed: DMEM+10%FBS after reaching the second filial generation; Unless stated otherwise, in the present invention, indication smooth muscle is P3-P5 generation.Rat smooth muscle cells (VascularSmoothMuscleCell, VSMC) is with 10
4the density kind in every hole, in 96 orifice plates, merges to 50%, removes serum and continue to hatch 24h, experimentally design subsequently, and add 0,2.5,5,10,20 and 40 μM of taxi driver brother respectively and receive after fragrant lactone C preincubate 24h, continuing to add serum to final concentration is 10%, puts into incubator.Take out after 24h, adopt crystal violet staining assay, observe the absorbance of each group cell at OD590 place.Logarithm probability diagram method calculates brother and receives the half-inhibition concentration (IC50) of fragrant lactone C.Be grouped as follows: Normal group: non-increase serum and brother receive the smooth muscle cell of fragrant lactone C; Model group: serum-concentration is the smooth muscle cell of 10%; Each dosage group: receive after fragrant lactone C preincubate 2h with brother, adding serum-concentration is 10%.
Experimental result: brother receive fragrant lactone C be dose-dependent inhibition Serum-induced VSMC propagation (each dosage group is compared with model group, p value is all less than 0.05), it is 86% that 40 μMs of taxi driver brothers receive the suppression ratio of fragrant lactone C, and brother receives fragrant lactone C, and to breed half-inhibition concentration to the VSMC of Serum-induced be 3.94 μMs/L.See accompanying drawing 1 and table 1.
Table 1. brother receives fragrant lactone C and suppresses the VSMC multiplication effect of Serum-induced
* represent and have statistical significance compared with model group, p<0.05
Experimental result shows: brother receives the vascular smooth muscle cell proliferation of fragrant lactone to Serum-induced, inhibited.
Embodiment 2. brother receive fragrant lactone C suppress PDGF-BB induction proliferation of vascular smooth muscle cells
Experimental technique: cell culture processes is with embodiment 1.Rat smooth muscle cells is with 10
4the density kind in every hole, in 96 orifice plates, merges to 50%, removes serum and continue to hatch 24h, experimentally design subsequently, add 0,1.25,2.5,5,10 and 20 μMs of taxi driver brothers receive fragrant lactone C preincubate 24h, add the somatomedin (bFGF100ng/ml of variable concentrations more respectively, EGF10 μ g/ml and PDGF-BB40ng/ml) continue to hatch, abandon cell conditioned medium after 24h, adopt crystal violet staining assay, observe the OD value of each group, and calculate the suppression ratio of each dosage group.Suppression ratio=1-[(model group OD value-blank value)-(each dosage group OD value-blank value)]/[(model group OD value-blank value)-(Normal group OD value-blank value)]; Logarithm probability diagram method calculates brother and receives the half-inhibition concentration (IC50) of fragrant lactone C.
Experimental result: brother receives fragrant lactone C and becomes the smooth muscle cell proliferation of dose-dependent inhibition PDGF-BB induction, and its half-inhibition concentration is 1.82 μMs/L, brother receives smooth muscle cell proliferation effect that fragrant lactone C induces bFGF and EGF without obvious inhibitory action.For the smooth muscle cell proliferation effect of these two kinds of growth factor-induced, brother's concentration received needed for fragrant lactone C50% suppression ratio is 10 times of PDGF-BB.Experimental result is shown in Fig. 2 and table 2.
Table 2. brother receives the inhibitory action of fragrant lactone C to different growth factor-induced VSMC multiplication effect
Experimental result shows: brother receives fragrant lactone C and suppresses smooth muscle cell proliferation effect, depend on somatomedin classification, namely, brother receive fragrant lactone C prioritizing selection suppress PDGF-BB induction smooth muscle cell proliferation, and to bFGF and EGF induction smooth muscle cell proliferation effect without obvious inhibitory action.
Embodiment 3. brother receive fragrant lactone C suppress PDGF-BB induction rat smooth muscle cells DNA synthesize
Experimental technique: cell culture processes is with embodiment 1.Rat smooth muscle cells is with 10
4the density kind in every hole is in 96 orifice plates, merge to 50%, remove serum to continue to hatch 24h, experimentally design subsequently, add 0, 0.25, 0.5, 1, 2 and 4 μMs of taxi driver brothers receive fragrant lactone C preincubate, adding PDGF-BB after 24h to final concentration is 40ng/ml, continue to hatch 24h, first 5 hours are terminated to experiment, each hole adds 10nMBrdu (aseptic process) respectively, continue to hatch to 24h, abandon cell conditioned medium, adopt PBS rinsing cell, Triton-100 penetration cell, paraformaldehyde is fixed, BSA closes and spends the night, little mouse-anti Brdu primary antibodie hatches 4h, PBST rinsing, FITC-labelling rabbit anti-mouse IgG bis-resists steps such as hatching, the last Brdu that observes under fluorescence inverted microscope absorbs positive rate (i.e. DNA synthetic ratio).Be grouped as follows: Normal group: do not add PDGF-BB and brother and receive the smooth muscle cell of fragrant lactone C; Model group: PDGF-BB final concentration is the smooth muscle cell of 40ng/mL; Each dosage group: receive after fragrant lactone C preincubate 24h with brother, adding PDGF-BB final concentration is 40ng/mL.DNA synthetic ratio=Brdu positive cell number (green fluorescence)/nucleus sum (blue-fluorescence) × 100%; Suppression ratio=1-(model group DNA synthetic ratio-each dosage group DNA synthetic ratio)/(model group DNA synthetic ratio-Normal group DNA synthetic ratio)
Experimental result: brother receive fragrant lactone C be dose-dependent inhibition PDGF-BB induction smooth muscle cell DNA synthesize.It is 31.34% (p<0.01) that 0.25 μM of brother receives fragrant lactone C suppression ratio, it is 61.82% (p<0.01) that 0.5 μM of brother receives fragrant lactone C suppression ratio, it is 89.77% (p<0.01) that 1 μM of brother receives fragrant lactone C suppression ratio, and it is 99.1% (p<0.01) that 2 μMs of brothers receive fragrant lactone C suppression ratio.The results detailed in Fig. 3 and table 3.
Table 3. brother receive fragrant lactone C suppress PDGF-BB induction DNA synthesis
* represent and have statistical significance compared with model group, p<0.05
* representative has statistical significance, p<0.01 with model group ratio
BrdU picked-up is the classical way judging that cell DNA synthesizes.Brdu can insert in the DNA double chain copied, and stably takes in daughter cell with cell division.Cell is after fixing and degenerative treatments, and available immunological method detects the content of BrdU in DNA, thus judges the multiplication capacity of cell.As simultaneously in conjunction with other cell marker (as nucleus dyestuffs such as DAPI, PI), double staining, can judge the kind of proliferative cell, growth rate, significant to research cyto-dynamics.Based on above-mentioned result of study, we have employed Brdu picked-up experiment further, receive fragrant lactone C whether have anti-PDGF-BB and induce VSMC proliferation function deeply to illustrate further brother.Result of study display brother receives fragrant lactone and suppresses the effect of smooth muscle cell proliferation of PDGF-BB induction relevant to cell cycle.
Embodiment 4. brother receive fragrant lactone C suppress PDGF-BB induction the rat smooth muscle cells cycle
Experimental technique: cell culture processes is with embodiment 1.Smooth muscle cell is with 5 × 10
6density plant in 100mm culture dish, to 50% merge, with plasma-free DMEM medium continue cultivate 48h.Award variable concentrations brother to receive and award 40ng/mlPDGF-BB after fragrant lactone C preincubate 24h and stimulate, continue to hatch 48h; Collecting cell supernatant, uses 0.25% trypsin digestion cell, collecting cell suspension, centrifugal 1000r/min × 5min, abandons supernatant.Fix 18h with 70% cold ethanol of 4 DEG C of pre-coolings, adjust afterwards cell concentration be 106/ml, PI dye 37 DEG C hatch 30min after carry out flow cytometer showed, the percentage rate of the shared total cell of each phase cell in application ModFitLT3.0 software analysis cell cycle; Calculate Proliferating antigen Ki67 (PI), PI=(S+G2/M)/(G0/G1+S+G2/M).Be grouped as follows: Normal group: do not add PDGF-BB and brother and receive the smooth muscle cell of fragrant lactone C; Model group: PDGF-BB final concentration is the smooth muscle cell of 40ng/mL; Each dosage group: receive after fragrant lactone C preincubate 24h with brother, adding PDGF-BB final concentration is 40ng/mL.
Low cytometric analysis is the different principle of the different times DNA content residing for cell, distinguishes the cell being in different phase.Cell cycle was made up of G0/G1 phase, S phase, G2/M phase, and the S phase is DNA synthesis stage, and S phase cellularity ratio increases, and reflection cell proliferation enlivens.Proliferating antigen Ki67 refers to and is in the ratio that S phase and G2/M phase cell sum account for total cell number, and it reflects the growth rate of this group of cells.Proliferating antigen Ki67 refers to and is in the ratio that S phase and G2/M phase cell sum account for total cell number, and it reflects the growth rate of this group of cells.Proliferation index evaluates ability of cell proliferation " goldstandard ".Our result display: compared with normal group cell, the smooth muscle cell G0/G1 phase percentage of cells of PDGF-BB induction obviously declines, and be 52.33%, normal group is 78.33%; Contrary, PDGF-BB group G2/M phase and S phase cell content, apparently higher than normal group, are respectively 14.15% and 33.47%, and Normal group G2/M phase and S phase cell content are respectively 7.09% and 14.57%; Compared with PDGF-BB group, it is that dose dependent raises G0/G1 phase cell content and reduces G2/M and S phase cell content simultaneously that brother receives fragrant lactone C, brother receives fragrant lactone C0.25 μM, 0.5 μM, 1 μM, 2 μMs and 4 μMs of group G0/G1 phase cell contents, be respectively 59.33%, 61.37%, 63.23%, 66.07% and 70.03%; G2/M phase cell content is respectively 9.06%, 5.53%, 8.65%, 8.04% and 11.33%; S phase cell content is respectively 31.63%, 33.13%, 27.4%, 25.9% and 18.67%.Normal group and PDGF-BB group proliferation index are respectively 21.67% and 47.67%; Brother receives fragrant lactone C0.25 μM, 0.5 μM, 1 μM, 2 μMs with 4 μMs of groups compared with PDGF-BB group, proliferation index obviously declines, and is respectively 40.67%, 38.63%, 36.77%, 33.93% and 29.97% (p<0.01).Result of study refers to Fig. 4 and table 4.
Table 4. brother receive fragrant lactone C suppress PDGF-BB induction VSMC proliferating cycle
* represent and have statistical significance compared with model group, p<0.05
* representative has statistical significance, p<0.01 with model group ratio
Experimental result shows, and brother receives the smooth muscle cell proliferation that fragrant lactone C can obviously suppress PDGF-BB to induce, and Proliferating antigen Ki67 obviously declines, brother receive fragrant lactone C can obviously by cell cycle arrest in the G0/G1 phase.
Embodiment 5. brother receive fragrant lactone C suppress PDGF-BB induction migration of vascular smooth muscle cells
Experimental technique: cell culture processes is with embodiment 1.Smooth muscle cell is with 5 × 10
4the density kind in every hole, in 24 well culture plates, merges to 80%, removes serum and spend the night.The aseptic rifle head of secondary daily 200uL draws "+" word in the middle of the hole, washes away the cell hiked up, take pictures under inverted microscope with normal saline, and awards variable concentrations brother and receive fragrant lactone C, and awarding 40ng/mlPDGF-BB after 2h stimulates; Take pictures under inverted microscope after 12 hours.Observe living cells to the situation of acellular zone migration, judge the transfer ability of cell.Migration area=observe initial acellular region area (the pixel)-acellular region area of observation terminal (pixel).Be grouped as follows: Normal group: do not add PDGF-BB and brother and receive the smooth muscle cell of fragrant lactone C; Model group: PDGF-BB final concentration is the smooth muscle cell of 40ng/mL; Each dosage group: receive after fragrant lactone C preincubate 24h with brother, adding PDGF-BB final concentration is 40ng/mL.
Experimental result: PDGF-BB induces 12 hours, significantly improve smooth muscle cell migration ability, showing as and observe initial acellular region area and observe the changing value of the acellular region area of terminal apparently higher than Normal group, is 3.12 times (p<0.01) of matched group, compared with PDGF-BB group, brother receives the smooth muscle cell migration that fragrant lactone C can obviously suppress PDGF-BB to induce, showing as and observing initial acellular region area with the changing value of the acellular region area of observation terminal is that dose dependent declines, brother receives fragrant lactone C0.25 μM, 0.5 μM, 1 μM, organize 2.37 times (p=0.46) that migration area is respectively Normal group for 2 μMs and 4 μMs, 2.48 times (p=0.3), 1.68 times (p<0.05), 1.48 times (p<0.05) and 1.34 times (p<0.05).Result of study refers to Fig. 5 and table 5.
Table 5. brother receive fragrant lactone C suppress PDGF-BB induction VSMC migration
* represent and have statistical significance compared with model group, p<0.05
* representative has statistical significance, p<0.01 with model group ratio
Experimental result shows, and in 1-4 μM of dosage range, brother receives the smooth muscle cell migration that fragrant lactone C obviously suppresses PDGF-BB induce, and in lower dosage range, brother receives fragrant lactone C the trend suppressing it to move.
Embodiment 6. brother receives fragrant lactone C and suppresses the synthesis of rat smooth muscle cells extracellular matrix
Experimental technique: rat smooth muscle cells is with 10
4the density kind in every hole merges in 96 orifice plates to 50%, remove serum to continue to hatch 24h, experimentally design subsequently, add 0,0.25,0.5,1,2 and 4 μM of taxi driver brother receives fragrant lactone C preincubate, adding PDGF-BB after 2h to final concentration is 40ng/ml, continue to hatch 24h, collecting cell supernatant, adopt ELISA method to detect the concentration of ICAM-1 and VCAM-1 in supernatant.Crystal violet staining assay, detects OD value, and each group of ICAM-1 and VCAM-1 concentration OD value is corrected.Be grouped as follows: Normal group: do not add PDGF-BB and brother and receive the smooth muscle cell of fragrant lactone C; Model group: PDGF-BB final concentration is the smooth muscle cell of 40ng/mL; Each dosage group: receive after fragrant lactone C preincubate 2h with brother, adding PDGF-BB final concentration is 40ng/mL.
Experimental result: PDGF-BB acts on 24 hours, significantly improve the concentration (p value is all less than 0.01) of extracellular matrix ICAM-1 and VCAM-1 in culture medium supernatant, prompting PDGF-BB is induction of the synthesis of smooth muscle cell extracellular matrix and release, and brother receives fragrant lactone C in 0.25-4 μM of dosage range, ICAM-1 and the VCAM-1 synthesis of obvious suppression PDGF-BB induction and release, compared with PDGF-BB induction group, brother receive fragrant lactone C0.25 μM group have reduce ICAM-1 and VCAM-1 release trend (p is respectively 0.07 and 0.57); 0.5 μM of group significantly reduces ICAM-1 and VCAM-1 release (p value is all less than 0.05); 1 μM of group has the trend (p is respectively 0.08 and 0.17) reducing ICAM-1 and VCAM-1 release; 2 μMs of groups significantly reduce ICAM-1 release (p<0.05) the trend (p=0.07) reducing VCAM-1 release simultaneously; 4 μMs of groups significantly reduce ICAM-1 and VCAM-1 release (p value is all less than 0.05).Result of study refers to Fig. 6 and table 6.
Table 6. brother receive fragrant lactone C suppress PDGF-BB induction adhesion molecule release
In form, data represent the ratio of each group of OD value and Normal group,
* represent and have statistical significance compared with model group, p<0.05
* representative has statistical significance, p<0.01 with model group ratio
Experimental result is pointed out, brother receive smooth muscle cell extracellular matrix that fragrant lactone C induce for PDGF-BB synthesis with discharge inhibited.
Embodiment 7. brother receives fragrant lactone C and suppresses PDGFR kinase activity.
Experimental technique: by variable concentrations (0,0.25,0.5,1,2 and 4uM) brother receives after fragrant lactone C at room temperature hatches 1 hour with restructuring PDGFR β and ATPmix, adds ADP-Glo reaction substrate, continue under room temperature to hatch 40 minutes, add kinase assay reactant liquor subsequently and hatch 30min.Chemoluminescence method is adopted to detect the luminous value of each group.
Experimental result: it is the kinase activity of dose-dependent inhibition PDGFR β that brother receives fragrant lactone C, the receptor phosphorylation level showing as PDGFR β declines, in dose-dependent inhibition kinase activation within the scope of 0.25-4 μM, suppression ratio is respectively 26.14%, 37.85%, 56.20%, 63.90%, 99.8%; Half-inhibition concentration (IC50) is 0.7626 μM.Result of study refers to Fig. 7 and table 7.
Table 7. brother receives fragrant lactone C to the inhibitory action of PDGFR kinase activity
Pdgf receptor has tyrosine kinase activity, after the effect being subject to its part PDGF-BB, the tyrosine kinase of receptor plays its zymogenesis, rapid autophosphorylation, and by this signal transmission to downstream signal transduction path, thus play its mitogen effect.ADP-Glo
tMthe principle of KinaseAssay be utilize pdgf receptor autophosphorylation consume ATP generate ADP, the latter regenerates ATP under certain condition, and by Ultra ?Glo
tMluciferase detects chemiluminescence, and luminous value is higher, and illustrate that newly-generated ATP is more, namely pdgf receptor phosphorylation level is higher, and pdgf receptor kinase activity is higher.The result of the present embodiment illustrates that brother receives fragrant lactone C and can directly act on pdgf receptor, suppresses its kinase activity, blocks its downstream signal transduction path, thus suppress the biological effect of PDGF-BB induction.
In sum, the present invention is first using serum as derivant, confirm that brother receives the smooth muscle cell proliferation of fragrant lactone C to Serum-induced and has stronger inhibitory action, by three kinds of somatomedin bFGF common in serum, EGF and PDGF-BB observes brother as derivant and receives the depression effect of fragrant lactone C to the smooth muscle cell proliferation effect of this three kinds of derivants induction, finds that brother receives the smooth muscle cell proliferation that fragrant lactone C Selective depression PDGF-BB induces; The present invention so by PDGF-BB as derivant, observe brother receive smooth muscle cell proliferation that fragrant lactone C induces PDGF-BB, migration, DNA synthesis and cell generation cycle effect and extracellular matrix synthesis and release effects, further illustrate brother and receive smooth muscle cell proliferation that fragrant lactone can suppress PDGF-BB induce, migration and extracellular matrix and synthesize.By to pdgf receptor kinase Activity determination, find that brother receives fragrant lactone C and can directly act on pdgf receptor, suppress its kinase activation, thus the smooth muscle cell proliferation of minimizing PDGF-BB induction, migration and substrate synthesis, be the novelty teabag being applicable to suppress vascular smooth muscle cell proliferation and migration.
Claims (6)
1. brother receives fragrant lactone C suppresses in the medicine of smooth muscle cell proliferation and/or migration, pharmaceutical composition and pastille material installation application in preparation as shown in Equation 1
2. application according to claim 1, is characterized in that: described smooth muscle cell proliferation and/or migration be comprise endothelial injury, vascular smooth muscle cell quantity that exogenous mechanical stimulus factor causes increases and/or the migration of position.
3. brother receives the application of fragrant lactone C in preparation prevention or treatment vessel wall thickening and/or the medicine of luminal stenosis cardiovascular and cerebrovascular disease, pharmaceutical composition and pastille material installation as shown in Equation 1, it is characterized in that, described vessel wall thickening and/or luminal stenosis cardiovascular and cerebrovascular disease are with smooth muscle cell proliferation and/or move as pathologic basis.
4. application according to claim 3, it is characterized in that, described vessel wall thickening and/or luminal stenosis cardiovascular and cerebrovascular disease are selected from atherosclerosis, disease that postangioplasty restenosis, hypertension, pulmonary hypertension, diabetic angiopathy cause.
5. application according to claim 4, it is characterized in that, described atherosclerosis is selected from the aortic aneurysm because atherosclerosis of aorta causes, because of the coronary atherosclerotic heart that coronary atherosclerosis causes, the angiostenosis caused because cranium brain is atherosis, cerebral blood supply insufficiency or local thrombus are formed or plaque rupture, the vascular dementia that fragment comes off in the ischemic cerebral apoplexy that causes and chronic cerebral ischemia causes, the renal artery thrombosis renal ischemic caused because of atherosclerotic renal artery stenosis the kidney failure finally caused, the acra blood caused because artery of extremity is atherosis also finally develops into intermittent claudication and gangrene for obstacle, and the dyspepsia to cause because of superior mesenteric atherosclerosis, intestinal hypotension, constipation and stomachache.
6. application according to claim 4, is characterized in that, the disease that described diabetic angiopathy causes is selected from diabetic nephropathy, diabetic renal papillary necrosis, diabetic neuropathy, diabetic foot.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410520677.8A CN105520935B (en) | 2014-09-30 | 2014-09-30 | Brother receives fragrant lactone C and inhibits the purposes of vascular smooth muscle cell proliferation and migration |
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| CN201410520677.8A CN105520935B (en) | 2014-09-30 | 2014-09-30 | Brother receives fragrant lactone C and inhibits the purposes of vascular smooth muscle cell proliferation and migration |
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| CN105520935A true CN105520935A (en) | 2016-04-27 |
| CN105520935B CN105520935B (en) | 2019-05-31 |
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| CN111184032A (en) * | 2020-01-10 | 2020-05-22 | 云南农业大学 | Goniothalamus affinis nematode killing extract as well as preparation method and application thereof |
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Non-Patent Citations (2)
| Title |
|---|
| K.M. CHAN ET AL: "Goniothalamin induces apoptosis in vascular smooth muscle cells", 《CHEMICO-BIOLOGICAL INTERACTIONS》 * |
| SI WANG ET AL: "Goniolactones A-F, Six New Styrylpyrone Derivatives from the Roots of Goniothalamus cheliensis", 《J. NAT. PROD.》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111184032A (en) * | 2020-01-10 | 2020-05-22 | 云南农业大学 | Goniothalamus affinis nematode killing extract as well as preparation method and application thereof |
| CN111184032B (en) * | 2020-01-10 | 2021-05-07 | 云南农业大学 | Goniothalamus affinis nematode killing extract as well as preparation method and application thereof |
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