CN108096304A - Purposes of the pill of Eight Treasures in prevention and/or treatment gastric cancer medicament is prepared - Google Patents
Purposes of the pill of Eight Treasures in prevention and/or treatment gastric cancer medicament is prepared Download PDFInfo
- Publication number
- CN108096304A CN108096304A CN201810008984.6A CN201810008984A CN108096304A CN 108096304 A CN108096304 A CN 108096304A CN 201810008984 A CN201810008984 A CN 201810008984A CN 108096304 A CN108096304 A CN 108096304A
- Authority
- CN
- China
- Prior art keywords
- treasures
- pill
- cell
- purposes
- gastric
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/32—Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/37—Digestive system
- A61K35/413—Gall bladder; Bile
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/55—Glands not provided for in groups A61K35/22 - A61K35/545, e.g. thyroids, parathyroids or pineal glands
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/58—Reptiles
- A61K35/583—Snakes; Lizards, e.g. chameleons
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/618—Molluscs, e.g. fresh-water molluscs, oysters, clams, squids, octopus, cuttlefish, snails or slugs
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Epidemiology (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Immunology (AREA)
- Virology (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- Biomedical Technology (AREA)
- Botany (AREA)
- Physiology (AREA)
- Endocrinology (AREA)
- Alternative & Traditional Medicine (AREA)
- Nutrition Science (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Marine Sciences & Fisheries (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Rheumatology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to Chinese medicine purposes fields.More particularly to application of the pill of Eight Treasures in preparing to treat treatment and/or prevention stomach cancer, gastric precancerous lesion drug.Pill of Eight Treasures of the present invention can be used to prepare anti-gastric cancer medicament, have good development prospect, belong to first public new application.The ingredient of the pill of Eight Treasures mainly includes cow-bezoar, snake gall, antelope's horn, pearl powder, Radix Notoginseng, Moschus.
Description
Technical field
The invention belongs to tcm fields, and in particular to pill of Eight Treasures answering in prevention and/or treatment gastric cancer medicament is prepared
With.
Background technology
Stomach cancer is one of common pernicious digestive system tumor, and Fujian is occurred frequently, is ranked the first in whole malignant tumours.It is most of
Late period has been in when patient is diagnosed, has had the characteristics that grade malignancy is high and the death rate is high.At present although curing gastric cancer has length
The progress of foot, system chemotherapy research are in progress, and targeted therapy research is even more in the ascendant, but clinical efficacy is unsatisfactory, and
Because its sky high cost is still difficult to extensive use.Therefore, it is current to find safer effective medicine to prevent stomach cancer
One of research emphasis and hot spot.
The limitation of Modern medical therapy stomach cancer so that there is " multicomponent ", the traditional Chinese medicine of " Mutiple Targets " action character to receive
Concern.
The content of the invention
Chinese patent drug pill of Eight Treasures is formed with the eight taste purification of tcm such as natural ox gallstone, snake gall, antelope's horn, pearl, Moschus, Radix Notoginseng,
There is clearing heat and detoxicating, promoting blood circulation and removing blood stasis, analgesic.This problem is intended inquiring into pill of Eight Treasures from tumor cell proliferation, apoptosis etc.
Influence to outgrowth in stomach cancer cell body, should for pill of Eight Treasures clinical anti-cancer to disclose the effect of pill of Eight Treasures anti-gastric cancer and mechanism
With theoretical foundation is provided, also stomach cancer is treated for theory of traditional Chinese medical science clearing heat and detoxicating, promoting blood circulation and removing blood stasis and theoretical foundation is provided.
It is an object of the invention to provide pill of Eight Treasures in prevention and/or treatment stomach cancer, gastric precancerous lesion drug is prepared
Purposes.
Specifically,
The purposes includes pill of Eight Treasures and is preparing the purposes in preventing gastric cancer medicament；
The purposes includes pill of Eight Treasures and is preparing the purposes in treating gastric cancer medicament；
The purposes includes purposes of the pill of Eight Treasures in the drug for inhibiting stomach cancer cell transfer or diffusion is prepared；
The purposes includes purposes of the pill of Eight Treasures in treatment and/or/prevention gastric precancerous lesion drug is prepared；
The gastric precancerous lesion is including atrophic gastritis etc..
Pill of Eight Treasures main component of the present invention includes cow-bezoar, snake gall, antelope's horn, pearl powder, Radix Notoginseng, Moschus；Preferably
The eight treasures (choice ingredients of certain special dishes) capsule of Xiamen Chinese Medicine Co., Ltd's production, Chinese medicines quasi-word Z10940006.
The present invention is the study found that pill of Eight Treasures inside and outside is respectively provided with the effect for significantly inhibiting Growth of Gastric；Stomach can be reduced
Cancer cell vigor；Play an important role of to induce apoptosis in gastric cancer；The cycle progression of stomach cancer cell can be blocked, retardance stomach cancer cell exists
The G0/G1 phases, so as to inhibit cell Proliferation；Inhibited to migration (transfer) ability of stomach cancer cell, pill of Eight Treasures can be used
In the drug for preparing treating cancer.
Research finds that pill of Eight Treasures has certain preventive and therapeutic effect to gastric precancerous lesion, and atrophic gastritis is stomach simultaneously
The important step of carcinogenesis, the present invention demonstrate pill of Eight Treasures to atrophic stomach by establishing atrophic gastritis model
Scorching therapeutic effect can be used so as to illustrate that it can improve gastric precancerous lesion symptom, elimination or mitigation mucosa infection, pill of Eight Treasures
In the active drug for preparing prevention stomach cancer generation.
Description of the drawings
Fig. 1 is block diagram of the pill of Eight Treasures to stomach cancer cell effect of vigor, compared with the control group,*P ＜ 0.05；
Fig. 2 is microphotograph (200 ×) of the pill of Eight Treasures to MGC-803, AGS stomach cancer cell morphology influence；
Fig. 3 is culture medium photo of the pill of Eight Treasures to stomach cancer cell Colony forming capacity；
Fig. 4 is the microphotograph (200 ×) that pill of Eight Treasures influences apoptosis in gastric cancer；
Fig. 5 is influence block diagram of the pill of Eight Treasures to AGS Growth of Gastric quantity, compared with the control group,*P ＜ 0.05；
Fig. 6 is influence block diagram of the pill of Eight Treasures to AGS gastric cancer cell cycles, compared with the control group,*P ＜ 0.05；
Fig. 7 is the microphotograph (100 ×) that pill of Eight Treasures influences stomach cancer cell scratch damage repair ability；
Fig. 8 is the microphotograph (200 ×) that pill of Eight Treasures influences stomach cancer cell transfer ability；
Fig. 9 is influence schematic diagram of the pill of Eight Treasures to stomach cancer cell transplantable tumor tumor volume；
Figure 10 is experiment mice photo in the influence experiment that pill of Eight Treasures grows Gastric Cancer Xenograft of Nude Mice；
Figure 11 is tumour photo in influence experiment of the pill of Eight Treasures to Gastric Cancer Xenograft of Nude Mice tissue.
Specific embodiment
Patent for a better understanding of the present invention, with reference to the embodiment content that the present invention is furture elucidated, but this hair
The content of bright patent is not limited solely to the following examples.
It elaborates with reference to the accompanying drawings and examples to the present invention.
Growth inhibition effect of 1 pill of Eight Treasures of embodiment to stomach cancer cell and the inhibitory action to stomach cancer cell transfer/diffusion
First, material and method
(1) experiment material
1. cell line：Stomach cancer cell line (MGC-803, AGS), the Shanghai Chinese Academy of Sciences.In Fujian University of Traditional Chinese Medicine's medical experiment
It is preserved in heart liquid nitrogen.
2. Naked mouse：SPF grades of BALB/C male nude mouses 40, weight (20 ± 2) g, animal quality certification number：SCXK (Shanghai)
2007-0005, purchased from Shanghai Slac Experimental Animal Co., Ltd..
3. drug：Pill of Eight Treasures (abbreviation BBD), Chinese medicines quasi-word Z10940006, manufacturer：Xiamen pharmaceutical factory of traditional Chinese medicine share is limited
Company, specification 0.3g/.
4. reagent
5. instrument
6. the configuration of common agents
(1) BBD solution：By pill of Eight Treasures powder, 25mg/mL is configured to respectively (for cell with PBS (phosphate buffer)
Experiment) and 40mg/mL (be used for zoopery) solution, ultrasonic dissolution is spare after 4 DEG C of storages with 0.22 μm of filter filtering.
(2) complete cell culture fluid (RPMI 1640)：1640 basic culture solutions of RPMI are configured to containing 10% tire ox blood
(FBS), 100U/mL penicillin and 100 μ g/mL streptomysins clearly.Culture solution is dispensed by every bottle of 100mL, 4 DEG C are kept in dark place.
(3) MTT liquid：Aseptically, 250mg MTT are weighed, are put into the centrifuge tube of 50mL, add 50mL 0.01M
PBS is made into the MTT solution of 5mg/mL, is shaken with vortex to after being completely dissolved, and is kept in dark place in 4 DEG C.
(4) cells frozen storing liquid：It is made into 7 parts of basic culture solutions, 2 parts of hyclones and l parts of DMSO, it is pre- under the conditions of 4 DEG C
It is cold.
(2) experimental method
1. cell experiment
1.1 cell culture
By stomach cancer cell line MGC-803, AGS be placed in containing 10% heat-inactivated fetal bovine serum (FBS), 100U/mL penicillin and
In the RPMI 1640 culture mediums of 100 μ g/mL streptomysins, in 37 DEG C, 5%CO2It is cultivated in the cell incubator of saturated humidity.Carefully
With 0.25% trypsase/EDTA digestion by 1: 3 secondary culture (note when born of the same parents' monolayer adherence grows to 80%~90% fusion：
It is frozen in time in cell cultivation process, convenient for conservation；When each cell reached for 7 generation, recovery cell again).
1.2 mtt assay detect cell viability
Mtt assay detects influence of the pill of Eight Treasures to stomach cancer cell vigor.Take the logarithm growth period MGC-803, AGS stomach cancer it is thin
Born of the same parents are digested with 0.25% pancreatin and collect cell.10%FBS, 100U/mL penicillin and 100 μ g/mL (are contained with DMEM culture solutions
Streptomysin) cell suspension is made, cell count is carried out, adjustment cell density is 1 × 105A/mL is inoculated in 96 well culture plates
In, per 100 μ l of hole, put after being cultivated in 37 DEG C of cell incubators for 24 hours (cell confluency degree reaches 50-60%), it is respectively plus different dense
Pill of Eight Treasures solution (final concentration is respectively 0,250,500,750 μ g/mL) is spent, totally 4 groups, every group sets 8 multiple holes.Pill of Eight Treasures intervention is trained
After supporting for 24 hours, the 100 μ l of MTT solution of 0.5mg/mL are added in per hole, 37 DEG C are incubated 4h, then inhale the liquid abandoned in each hole, add in
100 μ l/ holes of DMSO vibrate mixing, are placed at room temperature for 10min, crystallization are made fully to dissolve simultaneously mixing, in full-automatic microplate reader 570nm
Each group absorbance (i.e. A values) is measured, and cell viability is calculated by following equation：Cell viability (%)=(experimental group A values/right
According to a group A values) × 100%.
1.3 morphological observation
Pill of Eight Treasures intervention culture rear MGC-803 and ags cell metamorphosis for 24 hours are observed under inverted microscope.It takes and is in
The stomach cancer cell of exponential phase, by 1 × 105A/mL is inoculated in 6 well culture plates, per hole 2mL, is placed in 37 DEG C of titanium dioxide
Cultivated in carbon incubator, when cell confluency degree reaches 50~60%, add in various concentration pill of Eight Treasures solution (0,0.25,0.5,
0.75mg/mL) intervene for 24 hours, with inverted microscope observation cellular morphology variation and cell growth status, then with 500 μ L/ hole PBS
Clean cell 2-3 times, with inverted microscope 200 × under observed, taken pictures.
1.4 cell colony Forming abilities are analyzed
Using influence of the Colony forming experimental study pill of Eight Treasures to stomach cancer cell Colony forming ability.MGC-803 and AGS are thin
Born of the same parents are inoculated in 6 orifice plates, and 5 × 105Overnight incubation in 37 DEG C of cell incubators is put in a/hole, treats that cell confluency degree reaches 50~60%
When, add in various concentration pill of Eight Treasures solution (0,0.25,0.5,0.75mg/mL) intervene for 24 hours after, digest, neutralize again, centrifugation,
It is resuspended, counts, be inoculated with 1000 cells respectively in six orifice plates, change liquid within 2~3 days, after continuously cultivating 7~10 days, work as culture plate
When occurring the visible clone of naked eyes in hole, stop culture, supernatant, PBS cleaning 2~3 times are abandoned in suction.4% paraformaldehyde is added in fix
L0min, PBS cleaning is used 2~3 times after abandoning 4% paraformaldehyde, is dyed 20min with violet staining liquid, is sucked dyeing liquor, use PBS
Cleaning 3 times, observation Colony forming situation of taking pictures.
1.5Hoechst dyeing observation Apoptosis
The influence for detecting pill of Eight Treasures and being died to MGC-803 and ags cell tune is dyed using Hoechst33258.MGC-803 and
Ags cell is inoculated in 6 orifice plates, and 2.5 × 105A/hole, puts overnight incubation in 37 DEG C of cell incubators, and cell confluency degree reaches 50
When~60%, with various concentration pill of Eight Treasures solution (0,0.25,0.5,0.75mg/mL) intervene for 24 hours.Abandon supernatant, 1mLPBS cleanings 3
Time；L mL4% paraformaldehydes are added in, room temperature fixes l0min；4% paraformaldehyde is abandoned, with 1mL PBS cleaning 3 times；It is added in per hole
1mL Hoechst33258 staining solutions, at 37 DEG C, containing 5%CO2Cell incubator in be protected from light incubation, it is clear with PBS after dyeing
It washes cell 3 times, is then taken pictures (200 ×) with inverted fluorescence microscope observation.
1.6 cell quantities are analyzed
Using influence of the full-automatic cell calculating instrument analysis pill of Eight Treasures to Growth of Gastric.Take the logarithm the AGS in growth period
Cell inoculation is in 6 orifice plates, and 2 × 105A/hole, puts overnight incubation in 37 DEG C of cell incubators, and cell confluency degree reaches 50~60%
When, with various concentration pill of Eight Treasures solution (0,0.25,0.5,0.75mg/mL) intervene for 24 hours.Again digest, neutralize, centrifuge, be resuspended
Cell, after Trypan Blue, inFull-automatic cell calculating instrument carries out analysis of accounts to ags cell.
1.7 cell cycles were detected
Cell cycle (DNA) analysis is carried out using flow cytometer.By 1 × 105The ags cell of a/mL is inoculated in 25mL
Blake bottle in cellar culture for 24 hours after, respectively with the pill of Eight Treasures solution of various concentration (0,0.25,0.5,0.75mg/mL) administration
Intervention is handled.Continue culture and collect cell, adjustment cell concentration to 1 × 10 afterwards for 24 hours6A/mL, with 70% ethyl alcohol in 4 DEG C
It is fixed overnight, it is washed 2 times through PBS to the cold, is placed in RNAse (8 μ g/mL) and PI (10 μ g/mL) and is incubated 30min.It is thin with streaming
Born of the same parents' instrument detect AGS cell cycle distribution situation, and with ModfitLT Version 3.0 (Verity Software House,
Topsham) software is analyzed.
The healing analysis of 1.8 cell scratch damages
5 straight lines are drawn at each orifice plate bottom of 6 orifice plates with marker pen, every every 0.5cm or so.Stomach cancer cell AGS with 3.5 ×
105/ mL is inoculated in 6 orifice plates, and cell covers with after overnight incubation in incubator.After 20 μ L pipette tips high pressure sterilizations, at pipettor,
It holds pipettor and horizontal line cut is carried out perpendicular to plate bottom using certain dynamics.Old culture medium is sucked after having drawn, adds in PBS cleaning 3
It is secondary, wash away the cell under drawing.Add various concentration pill of Eight Treasures solution (0,0.25,0.5,0.75mg/mL), be placed in be inverted it is micro-
It takes pictures under mirror, places into incubator and cultivate.Again 5 visuals field is taken to take pictures (100 ×) at random under microscope after for 24 hours.
1.8Transwell cell migration assay
Using influence of the Transwell detection pill of Eight Treasures to ags cell transfer ability.Ags cell is inoculated in 6 orifice plates, and 2.5
×105Overnight incubation in 37 DEG C of cell incubators is put in a/hole, when cell confluency degree reaches 50~60%, with various concentration eight treasures (choice ingredients of certain special dishes)
Red solution (0,0.25,0.5,0.75mg/mL) intervene for 24 hours.Again digest, centrifuge, and base weight is cultivated with the RPMI 1640 of blank
It is outstanding, after Countstar is counted, add in serum free medium and adjust cell concentration to 2.5 × 105/ mL takes 200 μ L in containing base
Matter glue (Matrigel) or the Transwell upper chambers without matrigel (migration experiment), while add in 0.75mL in lower room and contain
The RPMI1640 culture mediums of 10%FBS, and be put in 37 DEG C of cell incubators and cultivate 12h.Microscopic observation cell is seen to whether there is and drop
The cell to get off.Liquid in cell is suctioned out, is put into the hole equipped with PBS and cleans, 750 μ L4% paraformaldehydes is added in and fixes
Aperture, upper chamber add in 200 μ L and fix 10min, the paraformaldehyde in cell is sucked, cell is put into added with 750 μ L crystal violets
Aperture in (before not being added in the hole containing complete medium), upper chamber add in 200 μ L crystal violets, dye 10min, use
PBS cleaning upper chamber, cotton balls are dried, after being dried after cell, under inverted microscope observe cell distribution and randomly select 5 visuals field with
200 × take pictures, it moves and attacks to the cell number of room under Transwell to evaluate carefully from Shi Gan Shang Transwell by observation
Bao Gan are moved and invasive ability.
2. zoopery
2.1 animal feedings are established with Transplanted tumor model
30 male nude mouses are placed in the laminar-flow rack in SPF grades of laboratories and are raised, freely activity, ad lib, drinking-water lead to
Wind, illumination is sufficient, and room temperature is controlled in (25 ± 1) DEG C, relative humidity 70%~80%.It is tested after adaptability raising 3d.It takes
The ags cell of exponential phase is digested with 0.25% pancreatin and collects cell, and concentration is made as 5 × 107The cell of a/mL hangs
Liquid is mixed with matrigel 1: 1.By 0.2mL/ (i.e. 5 × 106A/only), it inoculates to prepare at nude mice right fore armpit and move
Plant knurl model.
2.2 animal packets and administration
Transplanted tumor model：After mouse inoculation 4d, control group (physiological saline 10mL/kg), eight are randomly divided by tumorous size
Baodan low dose therapy group (abbreviation BBD low dose groups) (0.25g/kg), pill of Eight Treasures high-dose therapy group (abbreviation BBD high doses
Group) (0.5g/kg), every group 10.And give gastric infusion, and 1 time a day, successive administration 31d.(note：Testing the dosage used is
According to clinical pill of Eight Treasures dosage, the given low of mouse is converted into according to body surface area.)
2.3 animals are observed and weight, knurl cubing
Observation animal state daily；Every other day with electronics balance measurement weight, with vernier caliper measurement tumor size, it is measured
Major diameter (A) and vertical transverse diameter (B), by formula V=0.5 × A × B2Gross tumor volume is calculated, describes gross tumor volume growth curve.
2.4 transplantable tumor tumor weights measure
After animal is administered, neck execution is taken off immediately after taking blood, tumor mass is removed, is taken pictures with digital camera, use electronic balance
Measure tumor weight.
3. statistical procedures
All data are analyzed using SPSS22.0 software packages, as a result with mean ± standard deviationIt represents,
Compare between multi-group data using one-way analysis of variance, compare between two groups of data and examined using t.P<0.05 has for difference
Statistical significance.
2nd, experimental result
(1) cell experiment result
1. influence of the pill of Eight Treasures to stomach cancer cell vigor
The results are shown in Figure 1 by MTT, MGC-803 and ags cell through various concentration pill of Eight Treasures (0,0.25,0.5,0.75mg/
ML after) intervening for 24 hours, pill of Eight Treasures can inhibit the cell viability of MGC-803 and AGS, and certain dose dependent is presented, and have aobvious
The inhibition of work；Significant significant difference (* P are respectively provided with compared with the control group<0.05), the results showed that pill of Eight Treasures can be external
Play an important role of to inhibit Growth of Gastric.
2. influence of the pill of Eight Treasures to stomach cancer cell form
It is observed under inverted microscope, it turns out that blank control group MGC-803, ags cell kytoplasm are uniform, kernel understands,
Extensibility is good, well-grown, forms adherent spindle cell.With the increase of pill of Eight Treasures activity, MGC-803, ags cell
Number gradually decreases kytoplasms and becomes muddy, and how rounded cell is, and volume-diminished, refractivity is poor, and cell suspends, and comes off and dead.This reality
The multiplication of stomach cancer cell MGC -803, AGS can be inhibited by testing the results show pill of Eight Treasures, and with the increase of concentration and enhance, in apparent
Dose-effect trend.See Fig. 2.
The results are shown in Figure 3 for Cell colony formation assay, MGC-803 and ags cell through various concentration pill of Eight Treasures (0,
0.25th, 0.5,0.75mg/mL) after intervention processing for 24 hours, low, middle concentration group (0.25mg/mL, 0.5mg/mL) is compared with the control group
Its cell colony quantity formed does not have significant change, and high concentration group (0.75mg/mL) is significantly reduced, had compared with the control group
Apparent dose dependent.The result of study shows that pill of Eight Treasures has inhibitory action to the Colony forming ability of stomach cancer cell, into one
Step confirms that pill of Eight Treasures plays an important role of to inhibit Gastric Cancer MGC -803 and ags cell growth.
4. influence of the pill of Eight Treasures to apoptosis in gastric cancer
Observation Apoptosis is dyed using Hoechst, the results are shown in Figure 4, and MGC-803 and ags cell are through various concentration
Pill of Eight Treasures (0,0.25,0.5,0.75mg/mL) after intervention processing for 24 hours, cell (i.e. apoptotic cell) the gradually increasing of high dye is presented
It is more, show that pill of Eight Treasures plays an important role of substantially to induce MGC-803 and AGS apoptosis in gastric cancer, apparent dose-dependent effect is presented.
5. influence of the pill of Eight Treasures to stomach cancer ags cell growth population and cell cycle
Cell counts as shown in figure 5, ags cell through various concentration pill of Eight Treasures (0,0.25,0.5,0.75mg/mL)
After intervention processing for 24 hours, the cell quantity of pill of Eight Treasures processing group substantially reduces (p compared with the control group<0.05), there is significant agent
Amount relies on.The results are shown in Figure 6 for cell cycle distribution, ags cell through various concentration pill of Eight Treasures (0,0.25,0.5,0.75mg/
ML) after intervention processing for 24 hours, G0/G1 phase cell proportions dramatically increase, and S phases cell proportion and G2/M phase cell proportions substantially subtract
It is few, compared with the control group with significant difference (p<0.05), there is concentration dependant effect.It should be the result shows that pill of Eight Treasures has resistance
Effect of the indigestion cancer cell G1 phases to S phase processes makes stomach cancer cell be stuck in the G0/G1 phases, inhibits proliferation of human gastric cancer cell, makes stomach cancer
The Leukopenia of growth.As a result prompt pill of Eight Treasures that can play the work of inhibition Growth of Gastric by regulating and controlling gastric cancer cell cycle
With.
6. the influence that pill of Eight Treasures shifts stomach cancer ags cell
The migration of cancer cell is the important driving factors of metastases.Scratch damage reparation and Transwell experiments are letters
Victory measures the classical way of cell repair ability and travel motion ability.
Scratch damage reparation is whether observation in vitro cell grows (reparation) extremely central cut area, and the fortune of cell is judged with this
Dynamic transfer ability.Scratch damage reparative experiment is the results show that with the rise of pill of Eight Treasures concentration, compared with the control group, cut
Distance Shortened degree is smaller, and pill of Eight Treasures is prompted to can inhibit the scratch removal ability of ags cell, and increases its scratch removal with concentration
Ability declines, as shown in Figure 7.
Transwell is to reflect the side of cell migration ability by calculating in same time through the cell number of small pore membrane
Method, the cell number through small pore membrane is more, and transfer ability is stronger.Transwell the experimental results showed that, control group has more thin
Born of the same parents pass through small pore membrane, and after the intervention of various concentration pill of Eight Treasures, the cell quantity through small pore membrane gradually decreases, and makees in dose-dependant
With illustrating that pill of Eight Treasures can significantly reduce the transfer ability of stomach cancer cell AGS, as shown in Figure 8.
In short, scratch damage reparation and Transwell are the experimental results showed that pill of Eight Treasures has substantially stomach cancer cell transfer
Inhibitory action.
(2) results of animal
Transplanted Gastric Carcinoma nude mice model is prepared for using AGS stomach cancer cells are inoculated；Transplantable tumor is naked during the experiment
Death does not occur for mouse, and transplantable tumor success rate is 100%, and visible knurl body is grown after ags cell inoculates 3d；Each group experimental animal
Diet, activity, the state of mind and excretion situations such as be showed no exception.It was treated in pill of Eight Treasures (0.25g/kg and 0.5g/kg)
Cheng Zhong, the state and weight of animal show pill of Eight Treasures to mouse without obvious toxic-side effects without substantially changeing；Pill of Eight Treasures can be shown
The growth for inhibiting Imaging in Nude Mice Models Beared Gastric Carcinoma is write, compared with the control group with significant difference (P<0.05), the results showed that pill of Eight Treasures
Equally there is significant inhibitory action to Transplanted Gastric Carcinoma in vivo.The result is shown in Fig. 9-11.Therefore, pill of Eight Treasures can be used for preparing
Inhibit the drug of stomach cancer cell diffusion or transfer.
Therapeutic effect of 2 pill of Eight Treasures of embodiment to gastric precancerous lesion
First, material and method
(1) experiment material
1. experimental animal：Cleaning grade Wistar male rats 42, weight (200 ± 20) g are tested purchased from Shanghai Si Laike
Company of Animals Ltd..
2. drug：Pill of Eight Treasures (abbreviation BBD), Chinese medicines quasi-word Z10940006, manufacturer：Xiamen pharmaceutical factory of traditional Chinese medicine share is limited
Company, specification 0.3g/.Pill of Eight Treasures solution：By pill of Eight Treasures powder, the molten of 40mg/mL is configured to PBS (phosphate buffer)
Liquid, ultrasonic dissolution, high pressure sterilization are spare after 4 DEG C of storages.
(2) experimental method
Rat laboratory adaptability feed 7 days, room temperature control between 18-25 DEG C, show normal growth after fasting not
Taboo water 12 is re-weighed when small, with randomized and the random district's groups method of dividision into groups, rat is divided into blank group 8, model group 34, to mould
Type group is using MNNG free waters method cooperation acid suppression, irregular diet and NaTDC, ethyl alcohol administration by gavage joint modeling, the modeling
The foundation of method namely Chronic Atrophic Gastritis in Rat model is (referring to Liu Lei etc.《Combination method causes Chronic Atrophic Gastritis in Rat mould
The foundation of type》, Chinese General Practice, the 110th phase of volume 13).2 rats are randomly selected from model group, into
Row, which is cut open the belly, takes the situation of stomach observation gastric mucosa, and after confirming modeling success, 24 rats are randomly divided into model control group, pill of Eight Treasures
Low dose group (0.25g/kg), pill of Eight Treasures high dose group (0.5g/kg), every group each 8.
Specifically modeling method is：First MNNG with distilled water is formulated as the storing liquid of 1g/L concentration, configured weekly once,
It is protected from light, is stored refrigerated, the drinking bottle that black is protected from light paper is posted in loading, is freely drunk for rat.In addition to blank group rat, remaining
Add the ranitidine hydrochloride with 0.03% in each group rat word material, give hungry alternate nursing of satisfying, feed 2 days, fasting 1 day, feed 2
That afternoon after it is with 20mmol/L sodium deoxycholate solutions with dosage 10mL/kg gavages；That afternoon of fasting 1 day is used
40% ethyl alcohol 10mL/kg gavages, continue 16 weeks.Blank group gives normal saline gavage, common standard word material.In modeling
16 weekends randomly selected 2 rats from model group rats, after taking antrum and each one piece of body of stomach tissue, with routine hematoxylin one
Eosin stains, the gastric mucosa of 2 rats is observed after being fixed with 20% formalin, under light microscopic atrophy, and intestines occurs
Change with atypical hyperplasia, model is successfully, reproduced to confirm.
Administration：Pill of Eight Treasures is low after modeling 16 weeks, high dose group is respectively with pill of Eight Treasures aqueous solution 0.25g/kg, 0.5g/kg agent
Gavage is measured, one time a day, blank group, model control group give 10ml/kg physiological saline gavages, one time a day, during treatment group's medication
Between be 12 weeks.
HE stained preparations gather：It after putting to death rat, cuts open the belly and takes stomach, full stomach is taken out away from cardia and pylorus 1.5cm detachments, in stomach
Full stomach is splitted in big curved side, takes 0.2 × 2.0cm strip gastric tissues, is immediately placed in after being fixed for 24 hours in 4% paraformaldehyde solution, stone
Wax embeds, and standby HE is dyed, the variation of light Microscopic observation stomach lining.
2nd, experimental result
1st, ordinary circumstance and weight
Blank group rat expression freely, energy it is good, flexibly happiness is dynamic, fur gloss, and diet is normal；Model control group rat
Apathetic, tired in moving, the in disorder deficient gloss of fur, diet is reduced, and medication group rat has different degrees of activity and diet
Increase, weight gain, fur owes gloss, and general state is compared with blank group rat expression, energy is not good enough, and other performances are better than model
Control group.
Table 1：12 weeks weight of experimental rat change situation
Note：The * P ＜ 0.01 compared with blank group, the * * P ＜ 0.05 compared with model control group, * * * P ＜ 0.01.
2nd, stomach lining pathology
(1) visually observe
Blank group gastric wall in rats elasticity is good, and glossy, mucous membrane surface flat smooth, coating mucus is more, and mucous membrane is in powder
It is red；Pill of Eight Treasures low dose group and pill of Eight Treasures high dose group gastric wall in rats elasticity preferably, mucous membrane be in light red or kermesinus, pleat compared with
It is deep, there is a small amount of mucus to adhere to；Mould control rats stomach wall is relatively thin, and elasticity is poor, owes gloss, no mucus, mucous membrane in pale asphyxia or
Canescence, part mucous membrane appearance is dispersed in blutpunkte and fine grained sample changes.
(2) HE dyes micro- sem observation
Blank group：All gastric mucosa epitheliums are complete, and cellular morphology is homogeneous, and body of gland marshalling is close in tunica propria,
Sporadically appear a small amount of lymphocyte, and rat body of stomach and antrum portion have no apparent cell infiltration, have no dysplasia and enteric epithelium
Metaplasia and Mucosal atrophy lesion.
Model control group：The body of gland atrophy of stomach lining part even disappears, and partial rat body of gland occurs back-to-back or co-walled existing
As foveolae gastricae extends, and visible inflammatory cell infiltration in whole rat stomach lamina proprias, cell infiltration is as deep as muscularis mucosae, part
The visible lymphocyte aggregation of mucous membrane and lymph follicle are formed, blood vessel hyperplasia, and the antrum portion disorganized hyperplasia of body of gland is apparent, basal part
Glandular epithelial hyperplasia, the visible goblet cell of partial rat, nuclear hyperchromatism even double-core, all rats have moderate dysplasia
And intestinal metaplasia.
Pill of Eight Treasures low dose group：The arrangement of gastric mucosa body of gland is relatively neat, and cellular morphology is than more uniform, in tunica propria
Body of gland arrangement is relatively close more neat, and the boundary of Glandular Epithelium and Gland pipe is more clear, the infiltration of partial rat stomach body of gland visible inflammatory cell,
Cell infiltration is mainly limited to mucous membrane shallow-layer, and moderate dysplasia and intestinal metaplasia, Shao Shuo great occur in a small number of rats
There is slight or moderate Mucosal atrophy lesion in mouse, and situation substantially improves compared with model control group.
Pill of Eight Treasures high dose group：Gastric mucosa epithelium is complete compared with model group, and cellular morphology is than more uniform, tunica propria inner gland
Body arrangement is relatively close more neat, and sporadically appear a small amount of lymphocyte, and the boundary of Glandular Epithelium and Gland pipe is clear, and partial rat stomach body of gland can
See cell infiltration, cell infiltration is mainly limited to mucous membrane shallow-layer, and slight dysplasia and intestines occurs in only a few rat
There is slight Mucosal atrophy lesion in epithelial metaplasia, only a few rat, and recovery situation is better than pill of Eight Treasures low dose group.
The result shows that：Pill of Eight Treasures aqueous solution has improvement gastric precancerous lesion rat ordinary circumstance, improves weight, can be used for
Treat atrophic gastritis.
Before atrophic gastritis is the important step that stomach cancer occurs, therefore pill of Eight Treasures is to stomach cancers such as atrophic gastritis
Lesion has therapeutic effect, so as to reduce stomach cancer incidence, plays the role of pre- anti-cancer.
Certainly, those skilled in the art in the art are it should be appreciated that above-described embodiment is intended merely to illustrate this hair
It is bright rather than as limitation of the invention, as long as in the spirit of the present invention, conversion to above-described embodiment becomes
Shape will all fall within the scope of the claims.
Claims (7)
1. purposes of the pill of Eight Treasures in prevention and/or treatment stomach cancer, gastric precancerous lesion drug is prepared.
2. purposes according to claim 1, it is characterised in that：The purposes is pill of Eight Treasures in prevention gastric cancer medicament is prepared
Purposes.
3. purposes according to claim 1, it is characterised in that：The purposes is pill of Eight Treasures in treatment gastric cancer medicament is prepared
Purposes.
4. purposes according to claim 1, it is characterised in that：The purposes is preparing inhibition stomach cancer cell turn for pill of Eight Treasures
Purposes in the drug of shifting or diffusion.
5. purposes according to claim 1, it is characterised in that：The purposes is preparing treatment and/or/prevention for pill of Eight Treasures
Purposes in gastric precancerous lesion drug.
6. purposes according to claim 5, it is characterised in that：The gastric precancerous lesion includes atrophic gastritis.
7. according to the purposes any one of claim 1-6, it is characterised in that：The ingredient of the pill of Eight Treasures mainly includes ox
Huang, snake gall, antelope's horn, pearl powder, Radix Notoginseng, Moschus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810008984.6A CN108096304A (en) | 2018-01-04 | 2018-01-04 | Purposes of the pill of Eight Treasures in prevention and/or treatment gastric cancer medicament is prepared |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810008984.6A CN108096304A (en) | 2018-01-04 | 2018-01-04 | Purposes of the pill of Eight Treasures in prevention and/or treatment gastric cancer medicament is prepared |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108096304A true CN108096304A (en) | 2018-06-01 |
Family
ID=62218789
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810008984.6A Pending CN108096304A (en) | 2018-01-04 | 2018-01-04 | Purposes of the pill of Eight Treasures in prevention and/or treatment gastric cancer medicament is prepared |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108096304A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110420226A (en) * | 2019-09-18 | 2019-11-08 | 厦门中药厂有限公司 | Pill of Eight Treasures is preparing the purposes in the drug for preventing or treating disease relevant to IL-6 inflammatory factor storm |
| CN114272278A (en) * | 2020-09-28 | 2022-04-05 | 厦门中药厂有限公司 | Application of Babaodan in preparation of medicine for treating cisplatin-resistant gastric cancer |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104983764A (en) * | 2015-07-22 | 2015-10-21 | 厦门中药厂有限公司 | Application of Babaodan in preparation of drugs for treating pulmonary fibrosis |
-
2018
- 2018-01-04 CN CN201810008984.6A patent/CN108096304A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104983764A (en) * | 2015-07-22 | 2015-10-21 | 厦门中药厂有限公司 | Application of Babaodan in preparation of drugs for treating pulmonary fibrosis |
Non-Patent Citations (2)
| Title |
|---|
| 冯正权等: "八宝丹配合介入治疗热毒血瘀型原发性肝癌的临床观察", 《中华中医药学刊》 * |
| 李洪伦等: "八宝丹在恶性肿瘤治疗中的应用探索", 《滨州医学院学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110420226A (en) * | 2019-09-18 | 2019-11-08 | 厦门中药厂有限公司 | Pill of Eight Treasures is preparing the purposes in the drug for preventing or treating disease relevant to IL-6 inflammatory factor storm |
| CN114272278A (en) * | 2020-09-28 | 2022-04-05 | 厦门中药厂有限公司 | Application of Babaodan in preparation of medicine for treating cisplatin-resistant gastric cancer |
| CN114272278B (en) * | 2020-09-28 | 2022-12-09 | 厦门中药厂有限公司 | Application of Babaodan in preparation of medicine for treating cisplatin-resistant gastric cancer |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN115364176B (en) | Application of dendrobium-derived nano vesicles in preparation of medicines for preventing and treating atherosclerosis diseases | |
| CN108096304A (en) | Purposes of the pill of Eight Treasures in prevention and/or treatment gastric cancer medicament is prepared | |
| CN102048727B (en) | Application of formononetin in preparing of medicament for restricting angiogenesis | |
| CN103372076B (en) | Treat the medicine of disease and the application of Radix Sanguisorbae and/or Radix Sangusorbae extract that are caused by propionibacterium acnes | |
| CN105520934A (en) | Application of micheliolide dimethylamine | |
| CN104546822B (en) | The medical usage of epimedium aglucone | |
| CN102058570A (en) | Application of carnosic acid in preparing medicament for suppressing angiogenesis | |
| CN108714149A (en) | Purposes of the garden burnet active constituent in preparing anti-tumor drug | |
| TWI359669B (en) | ||
| CN106265619A (en) | DFMO or DFMO and Rhizoma Zingiberis Recens extract application in the medicine of the preparation esophageal carcinoma and the prevention of hepatocarcinoma and clinical treatment | |
| CN101283994A (en) | Application of plumbagin in preparing the medicine for preventing the blood vessel from regenerating | |
| CN103948581B (en) | Levocarnitine combines L-arginine and is preparing the application in treating diabetic retinopathy neurotrosis medicine | |
| CN115040528B (en) | Application of digitonin D and anti-tumor metastasis medicine | |
| CN101278925A (en) | Application of embellin in preparation of medicament for inhibiting angiogenesis | |
| CN102247349B (en) | The purposes of hepatic lesions caused by salvianolic acid A prevention and/or treatment diabetes | |
| US10004713B2 (en) | Uses of chlorogenic acid in the preparation of medicaments for treatment of oligodendroglioma | |
| TW201902498A (en) | Use of a pharmaceutical composition for the preparation of a medicament for anti-Helicobacter pylori | |
| CN113384590B (en) | Application of quercus acutissima acid in preparation of medicine for treating pancreatic cancer | |
| CN111920809B (en) | Application of benzothiidine and salt thereof in preparation of antitumor drugs | |
| CN107823536A (en) | It is a kind of to be used to prevent and treat Tibetan medicinal composition of hueppe's disease caused by pasteurella multocida and preparation method thereof | |
| CN116270489B (en) | Ginger stomach-activating granule and preparation method thereof | |
| CN111840286B (en) | Application of Keping and salt thereof in preparation of antitumor drugs | |
| CN107296896B (en) | Pharmaceutical composition for treating glioma and preparation method thereof | |
| TWI635867B (en) | Use of herbal composition in preparation drug for inhibiting tumor cell metastasis | |
| CN106074564A (en) | Ursolic acid application in preparing anti-hydatid drugs |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180601 |
|
| RJ01 | Rejection of invention patent application after publication |