CN110201017A - Arasaponin combines application of the tanshinone IIA in preparation prevention colorectal cancer drug - Google Patents
Arasaponin combines application of the tanshinone IIA in preparation prevention colorectal cancer drug Download PDFInfo
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Abstract
The invention discloses a kind of application of arasaponin joint tanshinone IIA in preparation prevention colorectal cancer drug, are related to technical field of Chinese medicine, the dosage of the arasaponin and tanshinone IIA are as follows: arasaponin 900mg；Tanshinone IIA 60mg.The present invention prevents the generation of mouse inflammatory colorectal cancer using traditional Chinese medicine monomer arasaponin joint tanshinone IIA, and exploring to COX-2 signal path is its crucial mechanism of action, to push traditional Chinese medicine in the development of tumor prevention cause, preventives treatment of disease for motherland's medicine and strong foundation is provided.
Description
Technical field:
The present invention relates to technical field of Chinese medicine, and in particular to a kind of arasaponin joint tanshinone IIA is pre- in preparation
Application in anti-colorectal cancer drug.
Background technique:
Colorectal cancer is the common malignant tumour in China, and the disease incidence of colorectal cancer rises 3~4% every year in the past 30 years.
World Health Organization's National Cancer research representative office (International Agency for Research on Cancer,
IARC the Globocan 2012) delivered estimates that CRC Standardized incidence rate in China's Mainland is 14.2/10 ten thousand, occupies the 75th, the world, marks
Change case fatality rate 7.4/10 ten thousand, occupies the 78th, the world.However, China's CRC disease incidence and dead number of cases but account for whole world morbidity respectively
With the 18.6% of dead total number of cases and 20.1%, the 1st, residence.It is reported according to National Cancer Center whole nation tumour registration data,
Shelter has the 3rd and the 5th of malignant tumour respectively for China city and rural area CRC disease incidence, and case fatality rate occupies the 4th respectively
With the 5th.
The colorectal cancer of correlative study discovery 85%-90% or more is and the adenoma from Colon and rectum adenoma evolving development
Mainly by scorching disease ﹑ alcohol stimulation caused by, Colon and rectum adenoma is considered being important the precancerous lesion of colorectal cancer, the U.S. it has been recommended that
Part population rule takes the generation of low-dosage aspirin prevention colorectal cancer and cardiovascular disease, but aspirin alimentary canal
Adverse reaction is clear, and is not approved as colorectal cancer Yi ﹑ secondary prevention medication in China.
Based on China, a large amount of crowds take the fact that Radix Notoginseng and Radix Salviae Miltiorrhizae prevention and treatment cardiovascular disease for a long time, and have correlation
Research confirms that arasaponin has the dependent interaction channel of similar aspirin inhibition tumour, and Radix Notoginseng, pellet with tanshinone IIA
Ginseng is clinically common traditional medicine classics medicine pair.Radix Notoginseng has effects that removing blood stasis and hemostasis, promoting blood circulation analgesic therapy；Radix Salviae Miltiorrhizae has promoting blood circulation
Menstruation regulating, stasis-dispelling and pain-killing, cool blood to disappear carbuncle, relieving restlessness are calmed the nerves effect, and the two has blood circulation invigorating efficacies.Chinese medicine thinks, syndrome of blood stasis and swollen
Tumor is closely related.According to research reports, due to such as Rg1 and Rb1 relative molecular mass of the effective component in arasaponin
Larger, such intestinal absorption is poor, and can improve the intestinal absorption rate of Radix Notoginseng after Radix Salviae Miltiorrhizae and Radix Notoginseng compatibility.Radix Notoginseng and Radix Salviae Miltiorrhizae are matched
The influence of 5 pairs of tanshinone IIAs is little, and pharmacokinetics behavior has no significant change.The present invention is demonstrate,proved by animal experiment study
Real, arasaponin and tanshinone IIA use in conjunction have the function of preventing colorectal cancer.
Summary of the invention:
To solve the above-mentioned problems, the present invention provides arasaponin joint tanshinone IIAs prevents Colon and rectum in preparation
Application in cancer drug.
The present invention provides arasaponin joint tanshinone IIAs in preparation prevention of inflammation correlation colorectal cancer drug
Application.
The dosage of the arasaponin and tanshinone IIA are as follows: arasaponin 900mg；Tanshinone IIA 60mg.
The present invention also provides a kind of pharmaceutical compositions, and using arasaponin and tanshinone IIA as active constituent, medicine is added
The medicament that acceptable auxiliary material is prepared on.
Numerous studies prove that arasaponin and ginseng II A of ketone not only play the role of protecting cardiovascular and cerebrovascular, more have multipath anti-
Cancer effect.
Arasaponin anticancer mechanism mainly has:
1, to the direct inhibition of tumour cell.1. having by the expression for lowering tumour cell cycle and the GAP-associated protein GAP that rises in value
The increment and growth of the inhibition colon cancer tumours of effect, can inhibit the activity of SW480 human colon cancer cell, in arresting cell cycle
S phase and G2/M phase, the synthesis of blocking dna.2. inhibiting DNA of tumor cell synthesis duplication, the study found that arasaponin energy will be thin
Intracellular growth was blocked in the G0/G1 phase, is inhibited the increment of human liver cancer cells Hep G2, is induced cell apoptosis.3. arasaponin energy
Inhibit cervical cancer Hela cells proliferation, by blocking cervical cancer Hela cells rapamycin target protein (mTOR) signal path,
MRNA translation efficiency is reduced, to inhibit protein translation initiation complex.4. the arasaponin of high concentration combines triphen oxygen
Amine can enhance tamoxifen to the antitumor action of breast cancer cell.
2, promote apoptosis of tumor cells.Arasaponin can make stomach cancer cell line MKN-28 Apoptosis and make cell block
In the G1 phase.Arasaponin can be substantially reduced Lung Adenocarcinoma A 549 Cell survival rate, show effect and time and drug concentration according to
Lai Xing.Notoginsenoside R can inhibit leukemia HL-60 cell Proliferation, while promote Apoptosis, with drug concentration increase
Effect enhancing.
3, tumor cell differentiation is induced.The study found that ginsenoside Rb2 has Hepatocellular carcinoma cell line in Radix Notoginseng
Induction of differentiation can significantly reduce alpha-fetoprotein secretion, increase albumin secretory volume.
4, anti-tumor metastasis: 1. pigment epidermal derived factors (pigment epithelium-derived factor,
It PEDF) is a kind of angiogenesis inhibitors, research shows that sanchinoside E1 is the selective agonist of estrogen receptor, ginseng
Saponin Rb1 can directly act on mankind's PEDF gene promoter, dramatically increase transcription, protein expression and the secretion of PEDF.Table
The blood vessel formation against function of bright sanchinoside E1 is derived from the specificity of PEDF, and ginsenoside RB1 is adjusted by erss
PEDF inhibits the tubular structure of endothelial cell to be formed.2. inhibiting the platelet aggregation of tumor cell induction and anti-tumor metastasis.
For blood under high viscosity and high coagulant state, the effect of tumour cell and blood platelet becomes very active, tumour cell by release tissue factor,
The activation of the induced platelets such as fibrin ferment and matrix metalloproteinase, while tumour cell stimulating platelet particle discharges Adenosine diphosphate
Glycosides promotes platelet activation, and tumour cell also passes through activation thromboxane A2 and promotes blood platelet in tumor cell surface aggregation, deformation
And degranulation, tumor cell surface are covered by blood platelet, are not easy to be identified by host immune monitoring system, are protected tumour cell not
It is destroyed by host immune cell, avoids being killed and being removed.Correlative study finds that the patient after rectal cancer surgery works as blood platelet
It counts and is lower than 350 × 109When/L, 5 years survival rates are up to 80%, and are more than the patient of this critical value, triennial deposit rate down to
37.5%, prompt thrombocythemia and poor prognosis in rectal cancer patient closely related.Studies have shown that arasaponin no matter
The platelet aggregation that tumor cell induction can be significantly inhibited in vitro in vivo has potential antitumor hematogenous metastasis ability.
Compared with aspirin, arasaponin inhibits the effect of platelet adhesion reaction more significant, and mechanism is to blood platelet COX-1/TXA2
The facilitation of the inhibition of access and Human Umbilical Vein Endothelial Cells COX-2/PGI2 access inhibits the adherency of blood platelet.
5, arasaponin is by inhibiting nuclear factor (NF)-kB signal pathway to play anti-inflammatory effect.(NF)-kB is extensive
Pathophysiological process in play regulating and controlling effect, be adjustable a variety of inflammatory reactions and immunogene expression, including phenomena of apoptosis.
Researches show that: arasaponin can lower the expression of NF-Kb, and then inhibit the secretion of inflammatory cytokine TNF, IL-1 β.
Tanshinone IIA anticancer mechanism mainly has:
1, tanshinone IIA prevents the accumulation of β-catenin in cell by the expression of COX-2 in inhibition people's colon-cancer cell,
To block Wnt/ β-catenin signal path, vascular endothelial growth factor (vascular endothelia growth is lowered
Factor, VEGF) expression, to reduce colorectal cancer angiogenesis.
2, by inhibiting AKT/m-TOR signal path, lowering Bcl-2 and raising the expression of Bax, tumour cell is promoted to wither
It dies.
3, promotion apoptosis of tumor cells is played by inducing the expression of tumor suppressor gene and influencing the transduction of signal pathway
Effect.
Chinese medicine prevents and treats disease by multicomponent, multiple target point, multipath, whole regulation body network, based on Ah
The anticancer mechanism of a woods is taken charge of, the invention demonstrates that arasaponin+tanshinone IIA high dose group can prevent inflammation by COX-2 approach
The generation of disease correlation colorectal cancer.
The beneficial effects of the present invention are: the present invention prevents mouse using traditional Chinese medicine monomer arasaponin joint tanshinone IIA
The generation of inflammatory colorectal cancer, and exploring to COX-2 signal path is its crucial mechanism of action, to push traditional Chinese medicine swollen
Tumor prevents the development of cause, preventives treatment of disease for motherland's medicine and provides strong foundation.
Detailed description of the invention:
Fig. 1 is mouse survival situation statistical chart；
Fig. 2 is modeling group knurl figure；
Fig. 3 is three red high group knurl figures；
Fig. 4 is normal gland figure；
Fig. 5 is intraepithelial neoplasia (cin) figure；
Fig. 6 is well-differentiated adenocarcinoma figure；
Fig. 7 is middle differentiation gland cancer figure；
Fig. 8 is arasaponin and/or tanshinone IIA tumour a situation arises statistical chart；
Fig. 9 is model group COX-2 strongly expressed figure；
Figure 10 is three red high dose group COX-2 weak expression figures.
Specific embodiment:
In order to be easy to understand the technical means, the creative features, the aims and the efficiencies achieved by the present invention, tie below
Specific embodiment is closed, the present invention is further explained.
By 88 Balb/c male mices, 11 groups, every group 8 are randomly divided by weight；Respectively AOM+DSS model comparison
Group, arasaponin+tanshinone IIA low dose group, arasaponin+tanshinone IIA middle dose group, arasaponin+Radix Salviae Miltiorrhizae
II A high dose group of ketone, arasaponin low dose group, arasaponin middle dose group, arasaponin high dose group, tanshinone
II A low dose group, tanshinone IIA middle dose group, tanshinone IIA high dose group, blank control group.Preceding 10 groups of intraperitoneal injections
AOM12.5mg/kg 1 time, equivalent 0.9%Nacl aqueous solution is injected intraperitoneally in blank control group, and first 10 groups start continuously certainly on the 5th day
By drinking 2.5%DSS aqueous solution 5 days, 3 Mondays circulation three is recycled totally.Blank control group normal water, every group feed it is high in fat
Diet.
It converts and measures by body surface area ratio, the clinical equivalent pharmaceutical quantities tanshinone IIA for calculating 20g or so mouse is
0.06mg, arasaponin 0.91mg, using this dosage as the low dosage of intragastric administration on mice, 2 times of amounts are middle dosage, and 4 times of amounts are
High dose, intragastric administration on mice volume are generally 0.2ml/10g, therefore each stomach-filling 0.4ml of 20g mouse, gavage daily 1 time, therefore are administered
Concentration is low dosage (Radix Salviae Miltiorrhizae 0.15mg/ml, Radix Notoginseng 2.28mg/ml), middle dosage (Radix Salviae Miltiorrhizae 0.30mg/ml, Radix Notoginseng 4.55mg/
), ml high dose (Radix Salviae Miltiorrhizae 0.60mg/ml, Radix Notoginseng 9.10mg/ml).
When 2nd period started DSS water circulation first day, starts simultaneously at medicine group and give arasaponin+pellet respectively daily
Join the basic, normal, high dosage of II A of ketone, the basic, normal, high dosage of arasaponin, the basic, normal, high dosage stomach-filling of tanshinone IIA, model group with
Blank control group gives the 0.9% parallel stomach-filling of physiological saline, until terminating for the 17th week.
Inflammatory correlation colorectal cancer mouse model is successfully produced using high fat diet+AOM and DSS, while applying high agent
It measures arasaponin joint high dose tanshinone IIA and feeds mouse to the neck that after the 17th week, breaks and put to death mouse, cut off mouse peritoneal,
Colon and rectum is separated with small intestine from cap end, until whole section of Colon and rectum is removed at anus, and longitudinally splitted along the intestinal tube longitudinal axis,
It cleans, examines record and occur and occur number, size and location whether there is or not tumour, calculate Tumor incidence, rear routine group
Dehydration is knitted, paraffin embedding is sliced, and HE dyeing, pathology diagosis is made a definite diagnosis.Using Transitional cell carcinomas (COX-2) antibody test mouse intestines
Road knurl, immunohistochemistry scoring: occurring brown yellow granule with cell cytosol and/or karyon, person is the positive.
Positive cell range is divided into 1-3 grades: no positive cell be 0 point, 1-10% remember 1 point, 11-50% remember 2 points, 50% with
Upper 3 points of note；Staining power is divided into 3 grades: 0 point of feminine gender note, and weakly positive remembers 1 point, and moderate positive remembers 2 points, and strong positive remembers 3 points, the two
The sum of be pathological tissues COX-2 score.As a result as follows: 1. survival rates
Arasaponin high dose group, tanshinone IIA Zhong ﹑ high dose group, arasaponin+tanshinone IIA Zhong ﹑ high dose
Group significantly improves the survival rate of inflammation associated colorectal cancer mouse, and difference is statistically significant (P < 0.05).With blank group ratio
Compared with arasaponin middle dose group and modeling group survival rate are lower, and difference is statistically significant (P < 0.05).
1.1 compared with modeling group
Red high group, group in pellet, group, the Radix Notoginseng high group death rate are below model group in three high group red, three pellets, difference has statistics
It learns meaning (P<0.05), remaining experimental group and model group comparing difference are not statistically significant (P>0.05), are shown in Table 1.
Statistical check (example, %) of 1 each group of table compared with the model group death rate
1.2 compared with blank group
Group, the model group death rate are above blank group in Radix Notoginseng, and difference is statistically significant (P < 0.05), remaining group and sky
White group is compared, and no significant difference (P > 0.05) is shown in Table 2.
Statistical check (example, %) of 2 each group of table compared with the model group death rate
In 1.3 groups relatively
Mouse death rate difference is not statistically significant (P > 0.05) between same dose group different disposal, is shown in Table 3.
The statistical check (example, %) that the death rate compares between different disposal in 3 same dose group of table
1.4 existence statistical charts
Different group mouse survival situation statistical charts are as shown in Figure 1, modeling group survival rate is minimum.
A situation arises compares for 2 mouse tumors
After experiment in the 17th week, mouse is dissected, the visible knurl of mouse intestinal is shown in Fig. 2 and 3.
Knurl send pathological examination, as the result is shown (HE dyes 10 × 10 times): mouse intestinal normal gland is shown in Fig. 4；Epithelium
Interior tumor becomes, and sees Fig. 5；Well-differentiated adenocarcinoma is shown in Fig. 6；Middle differentiation gland cancer, is shown in Fig. 7.
As the result is shown: dosage Zu ﹑ arasaponin is low in the high dosage Zu ﹑ tanshinone IIA of arasaponin+tanshinone IIA
Compared with model group, tumour substantially reduces dosage group, and difference is statistically significant (P < 0.05), is shown in Table 4-6, and table 7 shows blank
Group is compared with model group, and tumour substantially reduces, P < 0.05, and difference is statistically significant, it is believed that modeling is successful.
2.1 arasaponins+tanshinone IIA high dose group is compared with model group
Arasaponin+tanshinone IIA high dose group is compared with model group, and tumour substantially reduces, and difference has statistics
It learns meaning (P < 0.05), is shown in Table 4.It is believed that tumour is fewer than model group after arasaponin+tanshinone IIA high dose group is intervened.
4 arasaponins of table+tanshinone IIA high dose group and modeling group Tumor incidence situation (example, %)
Statistic=12.444, P=0.0004, P (exact)=0.0014
2.2 tanshinone IIA middle dose groups are compared with model group
Compared with model group, tumour substantially reduces (P < 0.05) tanshinone IIA middle dose group, and difference has statistics meaning
Justice is shown in Table 5, it is believed that tumour is fewer than model group after tanshinone IIA middle dose group is intervened.
5 tanshinone IIA middle dose group of table and modeling group Tumor incidence situation (example, %)
Statistic=7.2727, P=0.0070, P (exact)=0.0256
2.3 arasaponin low dose groups are compared with model group
Arasaponin low dose group is compared with model group, and tumour substantially reduces (P < 0.05), and difference has statistics
Meaning is shown in Table 6, it is believed that tumour is fewer than model group after arasaponin low dose group is intervened.
6 arasaponin low dose group of table and modeling group Tumor incidence situation (example, %)
Statistic=7.2727, P=0.0070, P (exact)=0.0256
2.4 blank groups are compared with model group
Compared with model group, tumour substantially reduces blank group, and P < 0.05, difference is statistically significant, is shown in Table 7, it is believed that
Modeling is successful.
7 blank group of table and modeling group tumor incidence situation (example, %)
Statistic=16.000, P=0.0001, P (exact)=0.0002
In 2.5 remaining each group group and the not statistically significant of comparison among groups
2.6 tumours a situation arises statistical chart, as shown in Figure 8:
3 immunohistochemical methods detect mouse knurl COX-2 horizontal expression
Mouse knurl COX-2 horizontal expression is detected using ImmunohistochemistryMethods Methods, as the result is shown: each dosage group ﹑ of tanshinone IIA
Each dosage group COX-2 level of arasaponin and modeling group comparing difference are not statistically significant (P > 0.05), compared with blank group,
Difference is statistically significant (P < 0.05).Arasaponin+tanshinone IIA high dose group COX-2 level is lower than modeling group and three
Seven total saposins+tanshinone IIA low dose group, arasaponin+tanshinone IIA middle dose group COX-2 level is lower than modeling group, poor
Different statistically significant (P < 0.05).Arasaponin+tanshinone IIA high dose group COX-2 level is lower than the high agent of tanshinone IIA
Amount group, difference are statistically significant (P < 0.05).COX-2 expression is shown in Fig. 9 and Figure 10 (HE dyes 10 × 10 times).
3.1 tanshinone IIA group results
Each dosage group of tanshinone IIA and blank group COX-2 level difference are statistically significant (P < 0.05), compare knot two-by-two
Fruit shows that group, low group red, modeling group COX-2 level are above blank group in high group red, pellet, and difference is statistically significant, remaining
Comparison among groups no significant difference, is shown in Table 8.With modeling group indifference, illustrate that Tanshinone ⅡA II cannot inhibit COX-2 horizontal.
Each dosage group COX-2 statistical check of 8 tanshinone IIA of table
3.2 arasaponin group results
Each dosage group of arasaponin and blank group COX-2 level difference are statistically significant (P < 0.05), compare two-by-two
The results show that arasaponin high dose group, arasaponin middle dose group, arasaponin low dose group, modeling group COX-2
Level is above blank group, and no significant difference between remaining each group is shown in Table 9.With modeling group indifference, illustrate the total soap of Radix Notoginseng
Glycosides cannot inhibit COX-2.
Each dosage COX-2 statistical check of 9 arasaponin of table
3.3 arasaponins+tanshinone IIA group result
Each dosage group of arasaponin+tanshinone IIA and blank group COX-2 level difference it is statistically significant (P <
0.05), comparison result is shown two-by-two, and the low middle high dose group of arasaponin+tanshinone IIA, modeling group COX-2 level are above
Blank group, arasaponin+tanshinone IIA high dose group COX-2 level are lower than arasaponin+tanshinone IIA low dose group
And modeling group, arasaponin+tanshinone IIA middle dose group COX-2 level are lower than modeling group, remaining each group difference is without statistics
Meaning is learned, is shown in Table 10.Illustrate that high dose group can inhibit COX-2 level in arasaponin+tanshinone IIA.
Each dosage group COX-2 statistical check of 10 arasaponins of table+tanshinone IIA
3.4 low dose group comparison results
Three low dose group COX-2 level differences are not statistically significant (P > 0.05), are shown in Table 11.
Each low dose group COX-2 statistical check of table 11
3.5 middle dose group comparison results
Three middle dose group COX-2 level differences are not statistically significant (P > 0.05), are shown in Table 12.
Each middle dose group COX-2 statistical check of table 12
3.6 high dose group comparison results
Three high dose group COX-2 level differences are statistically significant (P < 0.05), and comparison result shows that Radix Notoginseng is total two-by-two
Saponin(e+tanshinone IIA high dose group COX-2 level is lower than tanshinone IIA high dose group, remaining comparison among groups no statistical difference
Meaning (P > 0.05).It is shown in Table 13.Illustrate that arasaponin+tanshinone IIA high dose group is inhibiting COX-2 level with the obvious advantage.
Each high dose group COX-2 statistical check of table 13
In conclusion arasaponin joint tanshinone IIA high dose group and tanshinone IIA middle dose group can prevent inflammation
The generation of disease correlation colorectal cancer mouse tumor；And arasaponin joint tanshinone IIA high dose group can pass through inhibition
COX-2 approach carrys out the generation of prevention of inflammation correlation colorectal cancer mouse tumor.
The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention.The technology of the industry
Personnel are it should be appreciated that the present invention is not limited to the above embodiments, and the above embodiments and description only describe this
The principle of invention, without departing from the spirit and scope of the present invention, various changes and improvements may be made to the invention, these changes
Change and improvement all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appended claims and its
Equivalent thereof.
Claims (4)
1. arasaponin combines application of the tanshinone IIA in preparation prevention colorectal cancer drug.
2. arasaponin combines application of the tanshinone IIA in preparation prevention of inflammation correlation colorectal cancer drug.
3. application according to claim 1 or 2, it is characterised in that: the dosage of the arasaponin and tanshinone IIA
Are as follows: arasaponin 900mg；Tanshinone IIA 60mg.
4. application according to claim 1 or 2, it is characterised in that: be with arasaponin and tanshinone IIA for activity at
Point, the medicament that pharmaceutically acceptable auxiliary material is prepared is added.
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| CN112972513A (en) * | 2021-03-05 | 2021-06-18 | 中国药科大学 | Application of panax notoginseng saponins in preparation of medicine for treating colitis-related colon cancer |
| CN113768971A (en) * | 2021-09-27 | 2021-12-10 | 澳门大学 | Application of total tanshinone and total notoginsenoside in preparing medicine for preventing and treating colitis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112972513A (en) * | 2021-03-05 | 2021-06-18 | 中国药科大学 | Application of panax notoginseng saponins in preparation of medicine for treating colitis-related colon cancer |
| CN113768971A (en) * | 2021-09-27 | 2021-12-10 | 澳门大学 | Application of total tanshinone and total notoginsenoside in preparing medicine for preventing and treating colitis |
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Application publication date: 20190906 |