CN103845695B - A kind of compound stomach medicine of qi and activate blood circulation and preparation method thereof - Google Patents
A kind of compound stomach medicine of qi and activate blood circulation and preparation method thereof Download PDFInfo
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- CN103845695B CN103845695B CN201410044172.9A CN201410044172A CN103845695B CN 103845695 B CN103845695 B CN 103845695B CN 201410044172 A CN201410044172 A CN 201410044172A CN 103845695 B CN103845695 B CN 103845695B
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Abstract
A kind of compound stomach medicine of qi and activate blood circulation, by Radix Codonopsis 20g, Radix Astragali 30g, fructus amomi 9g, Radix Notoginseng 9g, RADIX ANGELICAE SINEINSIS 20g, fingered citron 10g, Rhizoma Chuanxiong 15g, curcuma zedoary 12g, 125 grams of meter/patch totally 200 pastes composition.Preparation method soaks above-mentioned medicinal material 0.5 hour, decocts 2 times, and for the first time plus 10 times are measured water, boil extraction 1.5 hours；Second plus 8 times of amount water, boil extraction 1.0 hours.6000 milliliters are concentrated into after merging extracting solution twice, filling to 500 mL of saline bottles, scarfing sterilizes, and refrigeration is i.e. spare.The present invention improves the degree of inflammation and atrophy degree of gastric tissue, mitigates mucosa tissue hyper-proliferative；Cytokine network is adjusted, serum TNF alpha active is improved, reduces IL 6, IL 8 generation；Inhibit key protein NF κ B expression and activation in gastric tissue NF κ B/I κ B alpha signal conduction paths；Inhibit the expression 1 of gastric tissue cyclinD, up-regulation p16 expression prevention gastric precancerous lesions.
Description
Technical field
The present invention relates to a kind of Chinese medicines and preparation method thereof, and in particular to a kind of stomach medicine of qi and activate blood circulation and its preparation side
Method.
Background technology
China is the High Risk For Gastric Cancer area, has 15~160,000 people to die of gastric cancer every year, case fatality rate occupy various malignant tumours
Forefront.Early diagnosis and early treatment are the current only effective measures for improving gastric cancer survival rate, the early diagnosis master of gastric cancer
The monitoring to gastric cancer variation early period is relied on anti-.Gastric cancer variation early period includes precancerous condition and precancerous lesion, Gastric precancerous condition
It is clinical concept, refers to gastric precancer diseases, the most common are atrophic gastritis；Gastric precancerous lesion (Precancerous
Lesions of gastric cancer, PLGC) it is pathological concept, including gastric epithelial intestinal metaplasia and dysplasia, it is chronic
Atrophic gastritis occurs together epithelium intestinal metaplasia or dysplasia is high risk factor that gastric cancer occurs.Studies have shown that viscous from normal stomach
Film-atrophic gastritis-intestinal metaplasia, Apoptosis are proportionate with hyperplasia；And from intestinal metaplasia-dysplasia-gastric cancer, Apoptosis
It is negatively correlated with hyperplasia, prompt the preliminary stage that occurs in gastric cancer, gastric epithelial cell group is extremely unstable, there is apoptosis and
Hyperplasia it is unbalance.Therefore, inquire into the pathogenesis of gastric precancerous lesion, effectively prevent its occurrence and development to reduce gastric cancer incidence have
How highly important meaning reduces the incidence of gastric cancer in early prevention and treatment and reverse PLGC, is the hot spot studied at present.
Cyclin D_1 gene (cyclinD1) is the shaping regulatory factor that the G1 phases enter the S phases in the cell cycle, is that the G1 phases are thin
The key protein of born of the same parents' proliferation signal, it is considered to be oncogene.Research finds cyclinD1, and there are gene magnification and eggs in gastric cancer
White overexpression, but it is unrelated with tumor differentiation degree, and this prompt cyclinD1 abnormal expression may be gastric cancer occurrence and development process
The molecular events of middle early stage.
P16 is first and is found to directly act on the cell cycle, inhibits fissional tumor suppressor gene.In cell Proliferation
In regulation and control, p16 albumen is competed with cyclinD1 and is combined with CDK4/6, is prevented, control cell division, and Cell differentiation inducing activity plays
Negativity adjustment effect.But when p16 protein delations, mutation and when causing the improper expression of p16 albumen, cell Proliferation will be out of control, causes
Tumour occurs.It is significantly lower than normal gastric mucosa and non-cancerous issue in expression of p 16 in gastric cancer positive rate.
Nuclear Factor-Kappa B (NF- κ B) is the important factor of controlling gene transcription, and Recent study shows its hair in tumour
It plays a significant role in raw, development and the regulation and control of Apoptosis.It with Apoptosis there are bidirectional modulation relationship, to it is numerous have it is anti-
The transcriptional expression of the albumen of apoptotic effect has the function of inducible up regulation, passes through the imbalance to apoptotic proteins and anti-apoptotic proteins
Expression modulation, protect cells from apoptosis threat.The anti-apoptotic function of NF- κ B can also by with tumor suppressor protein (such as
P53 etc.) between direct interaction be achieved.Many experiments confirm that NF- κ B are expressed sharp in stomach cancer cell and tissue
Living, the formation, development, infiltration, transfer with gastric cancer are closely related, and decide the prognosis of patient to a certain extent.But
Whether PLGC also has NF- κ B high expression, and how on earth is the relationship between cyclinD1, p16, all ten does not distinguish still at present
Chu.
PLGC belong to motherland's medicine " epigastric pain ", " feeling of fullness ", " it is noisy " and wait scopes, with taste deficiency of both qi and yin be this or it is simultaneous
The stagnation of the circulation of vital energy, blood stasis, the synthesis syndrome of simulataneous insufficiency and excessive.From the analysis of " prolonged illness must empty, pathogen usually intruding into collateral in protracted disease " theory of traditional Chinese medical science, in conjunction with its medical history,
Pathology reflects, it is believed that weakness of the spleen and the stomach is its main pathogenesis, and obstruction of collaterals by blood stasis is its pathology key.Chinese medicine and Chinese medicine compound prescription at present
Mostly is rested on the clinical observation stage to the research of PLGC, it is insufficient to the experimental study of Mechanism of TCM so that Chinese medicine treatment lacks
Weary reliable objective theoretical foundation lacks the further screening and verification of effective formula and drug.Yiqi Huoxue Recipe is mainly by Radix Codonopsis, Huang
Stilbene, fructus amomi, Radix Notoginseng, RADIX ANGELICAE SINEINSIS, fingered citron, curcuma zedoary composition.To apply the agreement side in more than 10 years in institute, to atrophic gastritis companion
Intestines clinical efficacy is definite, this research is point of penetration from NF- κ B, cyclinD1 and p16 relationships, inquires into Blood rheology to PLGC
The mechanism of action provides theoretical and experimental basis for Chinese medicine early stage intervention PLGC treatments.
Invention content
In view of the deficiencies of the prior art, the present invention provides a kind of compound stomach medicine of qi and activate blood circulation and preparation method thereof, improves
The degree of inflammation and atrophy degree of gastric tissue mitigate mucosa tissue hyper-proliferative；Cytokine network is adjusted, serum is improved
TNF-α activity reduces IL-6, IL-8 generation；Inhibit key protein NF- κ B tables in gastric tissue NF- κ B/I κ B alpha signal conduction paths
It reaches and activates；Inhibit the expression 1 of gastric tissue cyclinD, up-regulation p16 expression prevention gastric precancerous lesions.
The technical solution adopted in the present invention is：A kind of compound stomach medicine of qi and activate blood circulation, by including Radix Codonopsis 20g, Radix Astragali
30g, fructus amomi 9g, Radix Notoginseng 9g, RADIX ANGELICAE SINEINSIS 20g, fingered citron 10g, Rhizoma Chuanxiong 15g, curcuma zedoary 12g compositions, 125 grams of meter/patch 200 paste totally.
A kind of compound stomach medicine of qi and activate blood circulation, preparation method soak above-mentioned medicinal material 0.5 hour, decoct 2 times,
For the first time plus 10 times are measured water, boil extraction 1.5 hours；Second plus 8 times of amount water, boil extraction 1.0 hours.Merging is extracted twice
6000 milliliters are concentrated into after liquid, filling to 500 mL of saline bottles, scarfing sterilizes, and refrigeration is i.e. spare.
A kind of method of the compound stomach medicine of qi and activate blood circulation, middle dosage：21g/kg/d, by the low middle height of 1: 2: 4 ratio setting
Three dosage groups, then high dose group be：42g/kg/d, general liquid extract are all prepared into high dose group, and when administration moderately dilutes use,
By the dosage of 1mL/100g/d weight, i.e. 10mL/kg/d, crude drug content=(42g/kg/d)/(10mL/kg/d)=4.2g/
The crude drug content of mL is high dose group liquid extract concentration, medicinal extract amount=25000g/ (4.2g/mL)=5952mL, about 6000mL.
PLGC belongs to the scopes such as Chinese medicine " epigastric pain ", " feeling of fullness ", and ancient Chinese medicine doctor just recognizes its morbidity when earlier
Mechanism, clinical manifestation, and propose corresponding therapy and formula.PLGC is mostly because exopathogen is invaded or because of diet internal injury, emotional discomfort, is led
It causes liver-stomach disharmony, stomach Qi to become estranged, lead to drop dereliction of duty, stop in lucid yang failing to raise, turbid pathogenic factor, with the passing of time then the dysfunction of the spleen in transport, extravasated blood resistance network lead to stomach
Body, which loses, to be moistened, gastric gland atrophy.
《Treatise on the spleen and stomach》It says：" taste deficiency is all blood disease ".Sun Weifeng thinks that spleen deficiency causes the stasis of blood mainly by three aspects：When
Transformation failure of spleen causes the stasis of blood：Second is that changing source scarcity causes the stasis of blood；Third, the failure of the spleen to keep the blood flowing within the vessels causes the stasis of blood.Weng Weiliang thinks that disease is in regular period, development
To certain phase, the operation of qi and blood will be influenced, lead to disorder of qi and blood, generally the deficiency of vital energy, the stagnation of the circulation of vital energy, the circulation of vital energy in the wrong direction, the deficiency of Yin, cold solidifying, heat knot,
Phlegm is solidifying, dampness, body fluid deficiency liquid are few etc. can cause blood stasis, thus " all kinds of diseases and ailments all stasis of bloods ".SHAN Zhao wei points out that the interpretation of the cause, onset and process of an illness key of CAG is taste
The deficiency of vital energy, qi depression to blood stasis, diet is careless or disloyal, or is hindered by feelings will, undermines taste, deficiency of spleen-QI and stomach-QI with the passing of time occurs, and heresy enters blood network,
Gastric collateral stasis, insufficiency of vital energy and blood cause atrophy of gastric mucosa, and interpretation of the cause, onset and process of an illness key is in deficiency of spleen-QI and stomach-QI, blood-stasis internal-depression.
PLGC is with weakness of the spleen and the stomach for this, and taste consumption wound is root, and the stasis of blood is stagnant not to go, and compound is not given birth to, and poison is evil gradually deep, and heresy is gathered long very,
The empty or malicious or stasis of blood or more persons are mingled with, and are thusly-formed vicious circle, and with the passing of time stomach is become homeless foster, and mucous membrane is thinning, body of gland atrophy, whole cost
Disease forms " void ", the gesture of " stasis of blood " interior heap soil or fertilizer over and around the roots." stasis of blood " meets " two-hit theory " it is both new pathogenic factor, but also as cancer
The performance of preceding lesion.According to " treatinging deficiency with tonification, dissipate it actually ", the theory of " prolonged illness must be empty, pathogen usually intruding into collateral in protracted disease ", in view of this disease cause of disease disease
Machine it " void ", " stasis of blood ", therefore draft Blood rheology be the rules for the treatment of, side by Radix Codonopsis, Radix Astragali, Radix Notoginseng, Radix Angelicae Sinensis, fructus amomi, fingered citron, curcuma zedoary group
At.Specific side's solution：Ginseng, stilbene are monarch drug in a prescription altogether, and tonifying middle-Jiao and Qi, spleen-benefiting mind-tranquilizing, mutual reinforcement between is to use；Ministerial drug is made of Radix Notoginseng, Radix Angelicae Sinensis, is longer than
Replenishing and activating blood；Adjutant：Fructus amomi, fingered citron, curcuma zedoary.
Radix Astragali：It is sweet in flavor；Slightly warm in nature；Return lung, the spleen channel.It is swollen and other effects with tonifying Qi and lifting yang, invigorating qi for consolidating superficies, promoting pus discharge and tissue regeneration,《This
Grass meets original》Its " all void of energy the five internal organs " is called, is the key medicine of enriching spleen-QI.
Radix Codonopsis：It is sweet in flavor mild-natured；Returns spleen, lung channel.Work(is in help spleen and lung, nourishing generate fluid.《Book on Chinese herbal medicine is new again》：" tonifying middle-Jiao and Qi, and
Taste, relieving thirst and restlessness ".Transporting and transforming function of the spleen and stomach is normal, and qi and blood can be biochemical infinite, and medicine resultant force is so that " the vigorous spleen being able to resist against invasion of pathogen " is helped with Zhi Qiben
Just to get rid of evils.
Radix Notoginseng is sweet in flavor, slight bitter, warm-natured, Return liver, stomach, is classical good medicine promoting blood circulation and removing blood stasis, is both apt to removing blood stasis and hemostasis, detumescence is fixed
It bitterly, and can Xu-tonic；《Book on Chinese herbal medicine is looked for the truth》Cloud：" the bitter temperature of Radix Notoginseng smell, can be with its blood stasis of blood systemization.”
Radix Angelicae Sinensis：Sweet in flavor, pungent, slight bitter；Return liver, the heart, the spleen channel.With replenishing and activating blood, menstruction regulating and pain relieving, the effect of relaxing bowel.
《Book on Chinese herbal medicine is recorded through hundred kinds》Cloud：" when be classified as blood man must with medicine ... actually product are wanted in blood-nourishing.”
Fructus amomi is pungent, warm, returns spleen, stomach, kidney channel.The turbid promoting the circulation of qi of effectization, middle benefit gas.《Haigoushen》In say：" main cold air abdominal pain ...
Digest trough, warm taste." it is to be amusing to adjust stomach key medicine.
Fingered citron acrid, bitter, warm, returns spleen, stomach, liver, lung channel, effect qi-regulating and in,《Book on Chinese herbal medicine is just read》It says：" fingered citron, deficiency of YIN-blood
Person also dislikes its dry ear.
Curcuma zedoary：Acrid flavour, hardship；It is warm-natured；Return liver, the spleen channel.The work(of existing promoting the circulation of qi blood-breaking, Xiao Ji Zhi Tong can also dredge gas and QI invigorating,
When the empty people work(of the preferably simultaneous help of then curcuma zedoary.《Essentials of Materia Medica》Say its " addiction that disappears promoting menstruation, helping digestion of whetting the appetite, removing toxic substances analgesic.”《Book on Chinese herbal medicine warp
It dredges》Cloud：" taste element it is weak and without stagnant person, with anti-energy loss vital qi, make that food does not more disappear and taste benefit is weak ".As it can be seen that curcuma zedoary is especially suitable
For this patient with it is empty be this simulataneous insufficiency and excessive pathological state, Radix Astragali obtains curcuma zedoary tonifying Qi without in heap soil or fertilizer over and around the roots, curcuma zedoary obtains Radix Astragali, breaks through
Do not hinder just.Gastrointestinal pneumatosis is dispelled in interior abnormal fermentation, and with good fragrant digestion promoting function, and can preventing or arresting vomiting.
Modern pharmacology is thought：Radix Astragali can promote humoral immunity, cellular immunity, enhance body hypoxia-bearing and stress ability,
Rabbit blood Hemorheological Indexes are significantly reduced, property is identical as intensity and danshen injections；In addition, Radix Astragali also has protection stomach
Radix Astragali, Radix Angelicae Sinensis injection are carried out Zusanli point injection, can be obviously reduced space between cells, it is thin to restore epithelium by the effect of mucous membrane
Born of the same parents connect composite construction and effectively reverse the intestines and dysplasia of mucous membrane, the work with Inhibition test atrophic gastritis
Also have the function of antibacterial with, Radix Astragali and inhibits virus, cancer suppressing action.Radix Codonopsis energy Gastrointestinal motility adjustment, antiulcer, enhancing are immune
Function all has an impact excited and two kinds of nerve processes of inhibition.Radix Angelicae Sinensis has antithrombotic and improves lectin from hemolymph effect, improves blood
The effects that cycle, enhancing immunity of organism, anti-inflammatory and antalgic, scavenging activated oxygen, anti peroxidation of lipid.Radix Notoginseng has hematopoiesis function,
Humoral immune function can be improved, there are analgesic and anti-inflammatory effects, can obviously treat the atrophic lesion of gastric mucosa, and
Atypical hyperplasia and the intestinal metaplasia of galandular epithelium can be reversed, has and prevents function of tumor.Fructus amomi can enhance the function of stomach, promote
Digestive juice secretion can promote intestines peristalsis, can inhibit platelet aggregation；Curcuma zedoary：Modern pharmacological research confirms that oil of zedoary turmeric has anti-
Tumour, antimicrobial antiphlogistic, improves gastrointestinal smooth myokinesis at platelet aggregation-against, has extensive physiological activity, is especially inhibiting
Effect is unique in terms of tumour.Fingered citron has obvious inhibiting effect to intestinal smooth, can expand blood vessel, to too many levels immune function
It is obviously promoted effect, Peritoneal Macrophage Phagocytosis can be promoted.
Figure of description：
Fig. 1 is the influence that Blood rheology of the present invention expresses PLGC rat blood serum TNF-α；
Fig. 2 is the influence that Blood rheology of the present invention expresses PLGC rat blood serums IL-6；
Fig. 3 is the influence that Blood rheology of the present invention expresses PLGC rat blood serums IL-8；
Fig. 4 is influence of the Blood rheology of the present invention to PLGC rat gastric tissue NF- κ Bp65 protein expressions；
Fig. 5 is influence of the Blood rheology of the present invention to PLGC rat gastric tissue p16 protein expressions；
Fig. 6 is influence of the Blood rheology of the present invention to PLGC rat gastric tissue p16 protein expressions.
Specific implementation mode
With reference to the detailed embodiments of the present invention of embodiment：
A kind of compound stomach medicine of qi and activate blood circulation comprising Radix Codonopsis 20g, Radix Astragali 30g, fructus amomi 9g, Radix Notoginseng 9g, RADIX ANGELICAE SINEINSIS 20g,
Fingered citron 10g, Rhizoma Chuanxiong 15g, curcuma zedoary 12g, 125 grams of meter/patch 200 paste totally.Above-mentioned medicinal material is soaked 0.5 hour, is decocted 2 times,
For the first time plus 10 times are measured water, boil extraction 1.5 hours；Second plus 8 times of amount water, boil extraction 1.0 hours.Merging is extracted twice
6000 milliliters are concentrated into after liquid, filling to 500 mL of saline bottles, scarfing sterilizes, and refrigeration is i.e. spare.It is commented below by SD rats
Estimate the Chinese medicine effect.
(1) material
1. experimental animal
SD rats 64, cleaning grade, half male and half female, weight 90-110g, Shanghai Xi Pu-Bi Kai experimental animals Co., Ltd
It provides, credit number：SCXK (Shanghai) 2008-0016 is raised in Zhejiang University of Traditional Chinese Medicine's Experimental Animal Center.
2. experimental article
(1) experimental drug
N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) (lot number 50016) is company of Nuo Yang Bioisystech Co., Ltd
Product.
WEIFUCHUN PIAN (lot number 100105) Hu Qingyu Pharmaceutical Workshop pharmaceutcal corporation, Ltd product.
Yiqi Huoxue Recipe (Radix Codonopsis, Radix Astragali, fructus amomi, Radix Notoginseng, RADIX ANGELICAE SINEINSIS, fingered citron, Rhizoma Chuanxiong, curcuma zedoary) is by Hangzhou Chinese Medicinal Hospital's system
Suspension is made in agent room, per ml 3.1714g containing crude drug.
(2) main agents
Tumor necrosis factor-alpha (TNF-α) (lot number 11110403), interleukin-6 (IL-6) (lot number 11110404), Bai Jie
Plain -8 (IL-8) (lot numbers 11110405) are Shanghai Zhuo Kang biotech firms product.
Paraformaldehyde (lot number 20051208) Tianjin City Chemical Agent Research Institute product.
NF- κ Bp65 rabbit polyclonal antibodies (lot number G2010) are U.S.'s NeoMarkers products, working concentration 1: 50.
I κ B- α rabbit-antis (lot number G1411) are U.S.'s Santa Cruz products, and working concentration is 1: 50.
P16 (lot number I0110) is U.S.'s Santa Cruz products, and working concentration is 1: 50.
Cyclind1 (lot number J1810) is U.S.'s Santa Cruz products, and working concentration is 1: 50.
Envision two step methods immunohistochemical kit (lot number 00029661) is Dako companies of Denmark.
PBS phosphate buffers (ZLI-9062), solution final concentration：0.01M, citrate buffer (ZLI-9065),
Solution final concentration：0.01M, concentrated type DAB kit (ZLI-9032) (lot number 60882801), by Beijing Zhong Shan Golden Bridge biology skill
Art Co., Ltd produces.
DAB developing solutions (lot number 60119) provide for Beijing Bioisystech Co., Ltd of Zhong Shan Golden Bridge.
(3) laboratory apparatus, equipment
(2) method
The foundation of 1.PLGC rat models
Modeling group rat uses is added MNNG, a concentration of 167 μ g/mL in rat drinks water daily, while pressing every afternoon
The MNNG solution of 0.017mol/L concentration, 1ml/ mouse gavage attack, continuous 8 weeks, with normal rats forage feed.
2. animal packet and administration
64 rats are randomly divided into normal group 8 and modeling group 56, and it is big to put to death 10 modeling groups at random through modeling intervention
Mouse simultaneously causes gastric precancerous lesion model through proved by pathology, and it is high that remaining modeling group rat is randomly divided into model group 12, qi and activate blood circulation
Dosage group 12, qi and activate blood circulation low dose group 12, stomach answer spring group 10.Qi and activate blood circulation high and low dose group is given respectively daily
The Chinese medicine gavage of 1.6g/kg weight d and 0.8g/kg weight d is treated, and stomach, which answers spring group and gives stomach, answers spring suspension (1.23g/
Kg.d) gavage is treated, and model group is with the physiological saline gavage of equal capacity, co-continuous 12 weeks.Each group rat is observed during experiment
Ordinary circumstance.
3. collection of specimens
Processing rat, the anesthesia of 3% yellow Jackets took gastric tissue, were cut along greater curvature on an empty stomach morning next day after the last administration
It opens and makees gross examination of skeletal muscle, take lesser curvature side sinus body boundary gastric tissue, fixed with 4% paraformaldehyde, step by step dehydration of alcohol, dimethylbenzene is saturating
Bright, it is for use routinely to prepare paraffin section for paraffin embedding.
4. Testing index and method
(1) Histopathologic changes：Conventional H E dyeing and Masson trichrome stains, observe its atrophy of gastric mucosa degree and thickness
Degree.
(2) serum TNF-cc level：It is measured using ELISA method, is carried out in strict accordance with kit operating procedure.
(3 blood serum IL-6s, IL-8 are horizontal：Using measured by radioimmunoassay, carried out in strict accordance with kit operating procedure.
(4) in mucosa tissue NF- κ Bp65/I κ B α, cyclinD1, p16 albumen expression：Using immunohistochemistry
Envision methods detect.Main operational steps are as follows：Paraffin section dewaxing is to water, PBS rinsings, PBS after pressure cooker antigen retrieval
It rinses, PBS is rinsed after 3%H202 room temperatures 10min, and 37 DEG C of incubation 60min, PBS rinsings of primary antibody, the 37 DEG C of incubations of dropwise addition secondary antibody are added dropwise
60min；It is incubated 60min, PBS rinsings, DAB, which fully develop the color, to be terminated after 1~2min and fully wash, and haematoxylin is redyed, and is dehydrated simultaneously
Mounting；Wherein replace primary antibody as negative control using PBS.
5. diagnostic criteria
(1) gastric mucosa tissue pathological observation, with reference to the pathology in national chronic gastritis seminar National Consensus in 2000
Standard.
Chronic inflammation：Normally (0 point)：Mononuclearcell is no more than 5 per high power field.Slightly (1 point)：Chronic inflammation is thin
Born of the same parents are less and are confined to mucous membrane shallow-layer, are no more than the 1/3 of mucous layer.Moderate (2 points)：Chronic inflammatory cells comparatively dense is more than glutinous
The 1/3 of film layer, reaches 2/3.Severe (3 points)：Chronic inflammatory cells are intensive, occupy mucous membrane holostrome.
Atrophy：Slightly (1 point)：Intrinsic body of gland number reduces 1/3 no more than original body of gland, and most of body of gland still retains.In
It spends (2 points)：Intrinsic body of gland number reduces by more than 1/3, but is less than 2/3.Remaining body of gland irregular distribution.Severe (3 points)：Intrinsic gland
Body number reduces by more than 2/3, only remains a small number of bodies of gland, even disappears completely.
Intestinal gland metaplasia：Intestines part accounts for body of gland and the superficial epithelium gross area 1/3 is below for slight (1 point), 1/3~2/3
It is moderate (2 points), 2/3 or more is severe (3 points).
Dysplasia：(1) low dysplasia：It mostly occurs in mucomembranous surface, nucleus is arranged closely in substrate, nuclear staining
The intensive deep dye of matter, core increase, and Mitotic index is less, core be normally at cell half or 2P3 hereinafter, still keeping certain polarity.
Gland structure is slightly irregular, and form is slightly disorderly, seldom occurs back-to-back and co-walled.(2) high grade dysplasia：Cell pleomorphism is bright
It is aobvious, it is seen that pathological mitotic figure, core floats up to cellular apical, disorganized, forms cladding, partially or completely loses pole
Property.Nuclear hyperchromatism, kernel is big, is in blister.Gland structure is abnormal, has back-to-back, co-walled and phenomenon of sprouting more.Low atypical hyperplasia 1
Point, high grade dysplasia 3 divides, and low-height dysplasia 2 divides.
(2) Masson dyes criterion：Collagenous fibres take on a red color in blue, muscle, elastic fibers, and cellulose is in purplish red
Color, red blood cell are in Chinese red, and nucleus is in blue brown.
(3) immunohistochemistry detection cyclinD1, NF- κ Bp65, I κ B α, p16 result evaluation standards：ImmunohistochemistryResults Results are equal
It is observed under identical conditions, it is in brown yellow granule, p16 positive cells that NF- κ Bp65, I κ B α positive cells, which show as endochylema/karyon,
It is in brown yellow granule to show as karyon, and it is in brown yellow granule that cyclinD1 positive cells, which show as karyon,.Result evaluation standard：Sun
Sex index：That is (IOD areas are with practical ruler by IOD/1228800 (IOD areas are calculated with pixel unit) or IOD/72041.227
Very little μm of 2 units calculate), it is brown color with the immunohistochemistry slice positive region of DAB colour developings, positivity index indicates that total optical density is equal
The value of the photo is assigned to, i.e., the yellowing of captured photo, value is bigger, and photo is more yellow, and the tissue corresponding to photo is evaluated with this
Positive degree.
6. statistical analysis
It is counted using SPSS10.5 software packages, data mean ± standard deviationIt indicates, according to the property of data
One-way analysis of variance, non-parametric test etc. is respectively adopted in matter and research purpose.
Three, result
(1) rat ordinary circumstance
At the end of experiment, model group, qi and activate blood circulation low dose group respectively have 2 rats deaths；Qi and activate blood circulation high dose group and
Stomach, which answers spring group, respectively 1 rats death；Normal rats are without death.Rats death mainly because after gavage reflux enter esophagus suck lung
Portion leads to death.
Normal rats fur is closely personal, and color white light pool, two have god, and activity is quick, and appetite is more, and stool is in particle
Shape, tail color are light red.Rat model fur is fluffy and disorderly, matt, and irritability, appetite is less, and apathetic stool smell is smelly dirty, and matter is more
It is partially dilute or dry dilute uncomfortable.Qi and activate blood circulation high and low dose group, stomach answers spring group rat also has fur fluffy and disorderly, and appetite is reduced, irritability,
But degree is light compared with model group rats, and stool still shapes.
Rats in normal control group stomach lining pinkiness, glossiness is bright-coloured, mucosal fold rule out of shape, and surface is with viscous
Liquid, it is flexible.Model group rats stomach lining is in dark gray, and glossiness is dark and gloomy, and stomach wall is more normally organized thin, and mucosal fold is flat,
In the visible bleeding erosion point of 5 rat gastric tissues.Qi and activate blood circulation high and low dose and stomach answer spring group has improvement compared with model group, not
See bleeding erosion point.
(2) gastric mucosa of rat histological change
PLGC rat model gastric tissues change normal rats gastric mucosal cell marshalling under light microscopic, Glands morphology rule
Then, accidental inflammatory cell invades profit lamina propria.The intrinsic body of gland number of PLGC model group rats gastric mucosas is reduced, and the visible inflammatory of lamina propria is thin
Born of the same parents invade profit, and partially visible cell arrangement is disorderly, and Glands morphology is irregular, and 4 visible slight enterocyte metaplasias have no apparent
Dysplasia (see Fig. 1-3).
Compared with normal group, model group rats gastric tissue mucosal thickness significantly reduces (P ＜ 0.01)；After drug therapy with mould
Type group compares, and qi and activate blood circulation high dose group rat stomach tissue thickness is obviously improved (P ＜ 0.05), qi and activate blood circulation low dose group, stomach
Fu Chunzu rat stomach tissue thickness is slightly thick compared with model group, but no significant difference (P ＞ 0.05), beneficial between three pharmaceutical intervention groups
Spring group is answered compared with qi and activate blood circulation low dose group and stomach by gas promoting blood circulation high dose group rat stomach tissue thickness to be improved apparent (P ＜ 0.05).
Compared with normal group, model group rats gastric tissue degree of inflammation obviously increases (P ＜ 0.01)；QI invigorating after drug therapy
Promoting blood circulation high and low dose group, stomach answer spring group rat stomach tissue inflammation score degree (P ＜ 0.05, P ＜ substantially reduced compared with model group
0.01)；But comparing difference is without conspicuousness (P ＞ 0.05) between three pharmaceutical intervention groups and normal group and each drug.Normal group
Rat has no atrophy, and after drug therapy compared with model group, qi and activate blood circulation high dose group rat stomach Telatrophy scores journey
It spends substantially reduced (P ＜ 0.05), qi and activate blood circulation low dose group, Wei Fuchunzu rat stomach Telatrophy score degree difference without notable
Property (P ＞ 0.05), comparing difference is shown in Table without conspicuousness (P ＞ 0.05) between three pharmaceutical intervention groups and normal group and each drug
1。
The comparison that 1. each group rat stomach tissue inflammation degree of table, atrophy degree are scored
* P ＜ 0.05, * * P ＜ 0.01 compared with normal group；The Δ P ＜ 0.05 compared with model group, Δ Δ P ＜ 0.01；With benefit
Gas promoting blood circulation high dose group ratio P ＜ 0.05
(3) influence of the Blood rheology to PLGC rat blood serums TNF-α, IL-6, IL-8
Model group rats Serum TNF-α, IL-6, IL-8 level more normally organize reduction, and wherein IL-8 levels are substantially reduced (P
＜ 0.01)；Qi and activate blood circulation high and low dose group and stomach answer spring group rat blood serum IL-6 levels and are substantially reduced (P ＜ compared with model group
0.01, P ＜ 0.05), no significant difference (P ＞ 0.05) between three pharmaceutical intervention groups；Stomach answers spring group rat blood serum IL-8 water
It is flat to be shown in Table 2 compared with qi and activate blood circulation high and low dose group raising (P ＜ 0.05).
Influence of 2. Blood rheology of table to PLGC rat blood serums TNF-α, IL-6, IL-8
* P ＜ 0.05, * * P ＜ 0.01 compared with normal group；The Δ P ＜ 0.05 compared with model group, Δ Δ P ＜ 0.01；With benefit
Gas promoting blood circulation high dose group ratio P ＜ 0.05；With qi and activate blood circulation low dose group ratio ■ P ＜ 0.05
(4) influence of the Blood rheology to PLGC rat gastric tissue NF- κ Bp65, I κ B α protein expressions
Normal rats gastric tissue has a small amount of NF- κ Bp65 positive expressions, based on cytoplasm, few cell nuclear expression, carefully
Endochylema has a small amount of I κ B α positive expressions.Model group rats gastric tissue has a NF- κ Bp65 high expression, positive expression in cytoplasm and
In nucleus, gastric tissue I κ B α have high expression, and positive expression is in cytoplasm.Model group rats gastric tissue NF- κ Bp65, I κ B α are equal
Relatively normal group is significantly raised, and difference is significant (P ＜ 0.01)；Qi and activate blood circulation high dose group, stomach answer spring group rat gastric tissue NF-
κ Bp65 expression is substantially reduced (P ＜ 0.05) compared with model group, and I κ B α protein expressions are significantly raised (P ＜ 0.01) compared with model group；Three
Between medicine group, NF- κ Bp65 protein expressions group differences are without conspicuousness, and qi and activate blood circulation low dose group I κ B α protein expressions are compared with QI invigorating
Promoting blood circulation high dose group and stomach are answered spring group and are substantially reduced (P ＜ 0.01).See Fig. 4-5, table 3,4.
Influence of 3. Blood rheology of table to PLGC rat gastric tissue NF- κ Bp65 protein expressions
Art P ＜ 0.05, * * P ＜ 0.01 compared with normal group；The Δ P ＜ 0.05 compared with model group, Δ Δ P ＜ 0.01；With
Qi and activate blood circulation high dose group ratio P ＜ 0.05；With qi and activate blood circulation low dose group ratio ■ P ＜ 0.05
Influence of 4. Blood rheology of table to PLGC rat gastric tissue I κ B α protein expressions
* P ＜ 0.05, * * P ＜ 0.01 compared with normal group；The Δ P ＜ 0.05 compared with model group, Δ Δ P ＜ 0.01；With benefit
Gas promoting blood circulation high dose group ratio P ＜ 0.05, P ＜ 0.01；With qi and activate blood circulation low dose group ratio ■ P ＜ 0.05, ■ ■ P ＜
0.01
(5) influence of the Blood rheology to PLGC rat gastric tissue cyclinD1, p16 protein expressions
Normal rats gastric tissue has a small amount of cyclinD1 and p16 positive expressions, based on karyon, model group rats stomach group
Knitting has cyclinD1 and p16 high expression.Model group rats gastric tissue cyclinD1 and p16 more normally organize apparent increase (P ＜
0.01, P ＜ 0.05)；Qi and activate blood circulation high dose group and stomach are answered the expression of spring group rat gastric tissue cyclinD1 and are obviously dropped compared with model group
Low (P ＜ 0.01, P ＜ 0.05)；The p16 expression of qi and activate blood circulation high dose group rat gastric tissue is compared with model group apparent increase (P ＜
0.01)；Between three medicine groups, cyclinD1, p16 protein expression group difference are without conspicuousness.It is shown in Table 5,6 and attached drawing 4-5.
Influence of 5. Blood rheology of table to PLGC rat gastric tissue Cyclind1 protein expressions
* P ＜ 0.05, * * P ＜ 0.01 compared with normal group；The Δ P ＜ 0.05 compared with model group, Δ Δ P ＜ 0.01；With benefit
Gas promoting blood circulation high dose group ratio P ＜ 0.05, P ＜ 0.01；With qi and activate blood circulation low dose group ratio ■ P ＜ 0.05, ■ ■ P ＜
0.01
Influence of 6. Blood rheology of table to PLGC rat gastric tissue p16 protein expressions
* P ＜ 0.05, * * P ＜ 0.01 compared with normal group；The Δ P ＜ 0.05 compared with model group, Δ Δ P ＜ 0.01；With benefit
Gas promoting blood circulation high dose group ratio P ＜ 0.05, P ＜ 0.01；With qi and activate blood circulation low dose group ratio ■ P ＜ 0.05, ■ ■ P ＜
0.01
Gastric cancer is in the very high malignant tumour of whole world morbidity and mortality, and global nearly half gastric cancer is happened at me
State, is in rejuvenation trend in recent years, and case fatality rate occupy various malignant tumour forefront.Early diagnosis, early treatment are to improve at present
Unique effective measures of gastric cancer survival rate, early diagnosis rely primarily on the monitoring to gastric cancer variation early period.Therefore, before inquiring into gastric cancer
The pathogenesis of lesion effectively prevents its occurrence and development, becomes the emphasis of gastric cancer study on prevention, and have to reducing gastric cancer incidence
Highly important meaning.
(1) foundation of gastric precancerous lesion rat animal model
Animal mould is heavily dependent on for the active drug of research, the screening prevention of gastric precancerous lesion pathogenesis
The reliability and repeatability of type, the current animal model of gastric precancerous lesion rat animal modeling mainly have stomach lining to damage repeatedly at present
Hinder method, immunity modeling method, low dose of x-ray local irradiation method, in conjunction with carcinogenic comprehensive molding method, active immunity combination stomach
Mucosa injury method etc..
It is that current domestic scholars select most gastric precancerous lesion models wherein to combine carcinogenic comprehensive modeling method,
Japan Sug-imura in 1967 etc. applies MNNG administrations soluble in water, successfully induces the sdenocarcinoma of stomach of Wistar rats, is luring
During cancer, gastric mucosal lesion experienced from erosion, atrophy, the evolution of regenerative proliferation, dysplasia to canceration, these
Lesion substantially conforms to change seen in clinical CAG gastroscopes, also illustrates that the atrophy of gastric mucosa and dysplasia are seemingly experimental
There must be lesion during animal gastric cancer.It is established with the complex method that Methyl nitro nitrosoguanidine (MNNG) is basic mutagens
Animal model.Hereafter, many scholars improve on the basis of the method, with copy more meet Human Gastric carcinoma before lesion mould
Type.
The research of early period is reported and combined to this research bibliography, selects MNNG carcinogens, is added in rat drinks water dense
Degree is the MNNG of 167 μ g/mL, while pressing the MNNG solution of 0.017mol/L concentration every afternoon, 1ml/ mouse gavage attack, even
After 8 weeks continuous.For Histopathology as it can be seen that normal rats gastric mucosa thickness is normal, epithelium is complete, and epithelial cell and body of gland arrangement are whole
Together, regular, the boundary of Glandular Epithelium and Gland pipe is clear；Model group rats gastric mucosa is thinning, and intrinsic body of gland number is reduced, the visible inflammation of lamina propria
Property cell invasion, partially visible cell arrangement is disorderly, and Glands morphology is irregular, shows that quoting MNNG for a long time can be such that rat stomach sticks
Envelope barrier is destroyed, and rotten to the corn, atrophy and part intestines are caused.Although part visible cell is disorganized, Glands morphology is not
Rule has no apparent dysplasia, if the extension modeling time can lead to dysplasiaMultiple gavage easily causes to be intubated gavage
Relevant complication increases the cost of experiment, how to reduce the death rate if rats death rate increasesBe worth we from now on into
One step is furtherd investigate.
(2) Blood rheology induces MNNG the influence of gastric precancerous lesion gastric mucosa degree of inflammation, atrophy degree
It is sheet, or the simultaneous stagnation of the circulation of vital energy, blood stasis, the synthesis disease of simulataneous insufficiency and excessive that PLGC, which is with taste deficiency of both qi and yin in motherland's medicine,
Group.From " prolonged illness must be empty, pathogen usually intruding into collateral in protracted disease " theory of traditional Chinese medical science analysis, reflect in conjunction with its medical history, pathology, it is believed that weakness of the spleen and the stomach is it
Main pathogenesis, obstruction of collaterals by blood stasis are its pathology key.Yiqi Huoxue Recipe mainly by Radix Codonopsis, Radix Astragali, Radix Notoginseng, Radix Angelicae Sinensis, fructus amomi, fingered citron,
Curcuma zedoary forms.Radix Codonopsis, Radix Astragali tonifying middle-Jiao and Qi control spleen deficiency originally；Radix Notoginseng, Radix Angelicae Sinensis blood circulation promoting and enriching；It is amusing with stomach in fructus amomi promoting the circulation of qi tune
Help ginseng, stilbene strengthening the spleen and stomach；Fingered citron qi-regulating is in；Curcuma zedoary promoting the circulation of qi blood-breaking disperse accumulation.Full side is mutually coordinated, play altogether Yiqi-Huoxue-Huayu it
Effect.
The study find that normal rats gastric mucosal cell marshalling under light microscopic, Glands morphology rule, accidental inflammatory are thin
Born of the same parents invade profit lamina propria.The intrinsic body of gland number of PLGC model group rats gastric mucosas is reduced, and the visible inflammatory cell of lamina propria invades profit, partly may be used
See cell arrangement disorder, Glands morphology is irregular, and 4 visible slight enterocyte metaplasias have no apparent dysplasia.Model
Group rat stomach tissue adherence thickness and more normal group of degree of inflammation significantly reduce；After drug therapy compared with model group, QI invigorating is lived
Blood high dose group rat stomach tissue thickness is obviously improved.Model group rats gastric tissue degree of inflammation, which is more normally organized, obviously to be increased；Medicine
It is substantially reduced compared with model group to answer spring group rat stomach tissue inflammation score degree for qi and activate blood circulation high and low dose group, stomach after object treatment.Just
Often group rat has no atrophy, and after drug therapy compared with model group, qi and activate blood circulation high dose group rat stomach Telatrophy counts
Divide degree substantially reduced.Show that Blood rheology induces the hyper-proliferative of gastric precancerous lesion gastric mucosa cell to have suppression MNNG
It makes and uses, can partly reverse the Histopathologic changes of stomach lining, reduce the release of inflammatory cell, improve mucosa nutritive shape
Condition mitigates gastric mucosa tissue degree of inflammation and atrophy degree, mitigates gastric mucosa tissue damage.
(3) TNF-α, IL-6, IL-8 influence of the horizontal and Blood rheology to it in PLGC rat blood serums
It is at present to think and infect and be immunized related about the pathogenesis of PLGC and indefinite more.As important inflammation
It may relate to the pathologic process of PLGC with TNF-α, IL-6 and the IL-8 of the immune-mediated factor.TNF-α is specific immune response
A kind of important medium between inflammatory reaction plays an important role in generation, the evolution of inflammation, be Chemokines and
Activity factor, suitable TNF-α may participate in the activated inflammatory reaction of immunocyte, have anti-infective, antiviral and antitumor work
With；And excessive TNF-α then enhances wall of micrangium permeability, by raising Some Adhesion Molecules on Endothelial Cells inducing endothelial cell
Platelet activating factor and chemotactic cytokine IL-8 are generated, changes mucous membrane vascular endothelial cell reactivity, causes in capilary
Blood coagulation causes cytoclasis, lysosome to leak out, and inflammatory mediator release causes the damage of stomach lining endothelial cell, blood flow is made to subtract
It is few, and Gastric Mucosa Blood Flow is most important in the defense reaction of gastric mucosal barrier, to cause mucosal lesion.Meanwhile it is also
It can enhance it with chemotactic neutrophil leucocyte and swallow killing ability, the infiltration of enhancing inflammation local lymphocytes and proliferation, to induce
A series of inflammation changes, and aggravates mucosal inflammation reaction [8].TNF-α can stimulate body generate a series of inflammatory cell because
Sub (IL-1, IL-6 and IL-8) generates " water fall effect ", can induce Apoptosis, and in addition TNF-a can be in IL-6 presences, induction
Fibrin ferment is formed, and endothelial cell surface is made to become promoting coagulation, mucous membrane thin vessels shape on inflammatory basis from anticoagulation state
At microthrombus, lead to mucous membrane microcirculation disorder, weakens gastrointestinal tract mucous barrier function, while nuclear factor can be activated again
NF- κ B etc., the latter, which has, promotes cell survival, proliferation and differentiation, anti-apoptotic activities, especially in, severe atrophic gastritis
Pathological change in there may be the immunologic mjuries caused by TNF-α, in vivo expression also influence tumour generation, hair
Exhibition.
IL-6 is mainly secreted by monocytes/macrophages, and when inflammation is with stress reaction, it is a variety of to can induce body generation by IL-6
Acute phase protein, be acute phase reactant synthesis important medium, meanwhile and Organism immunoregulation important cytokine,
T, the proliferation of B cell, the various biological effect of differentiation and NK cells and macrophage are participated in, related IL-6 influences cancer cell
Report is different, and the IL-6 that tumour cell generates may be used as a kind of autocrine or paracrine growth factor, with cell table
The special membrane receptor effect in face, to the growth of tumour cell, the simultaneous phenomenon of tumor patient, the pathologic process of disease and the state of an illness
Severity has an impact.
IL-8 mainly by lipopolysaccharides or is swollen by monocytes/macrophages, histocyte (endothelial cell, fibroblast) etc.
The stimulations such as tumor necrosis factor (TNF-α), IL-1 generate, and are a kind of main chemotactic factor (CF)s.It is to neutrophil leucocyte, basophilla grain
Cell and T cell have chemotaxis, and can promote the activation of neutrophil leucocyte so that neutrophil leucocyte is locally poly- in gastric mucosa
Collect and release superoxides and lysosomal enzyme, its these biological activities play important work during resisting pathogen
With, but the damage [11] of mucosa tissue is also caused simultaneously.Research find IL-8 expressed in the antrum that Hp infects compared with
It is obviously increased in the antrum of no Hp infection, and IL-8 levels increase [12] with the exacerbation of degree of inflammation.
The study find that model group rats Serum TNF-α, IL-6, IL-8 level more normally organize reduction, wherein IL-8 is horizontal
It is substantially reduced (P ＜ 0.01)；Qi and activate blood circulation high and low dose group and stomach are answered spring group rat blood serum IL-6 levels and are obviously dropped compared with model group
Low (P ＜ 0.01, P ＜ 0.05), no significant difference (P ＞ 0.05) between three pharmaceutical intervention groups.Show PLGC generation and
Stomach lining poor nutritional, epithelial repair obstacle and immunity of organisms are lowly related caused by conversion may lack with TNF-α.Through benefit
After gas Huoxue Therapeutic Method, gastric mucosa inflammation and atrophy degree are substantially reduced, and serum TNF-cc level increases, and prompt promotes
TNF-α secretion increases, it may be possible to which it reverses the proliferative disorder of PLGC stomach linings, to block it to gastric cancer transforming mechanism of action
One of.But the effect of TNF-α is not adjusted yet to close to normal level, prompt this research that the application of the medicine may have not yet been reached
Best dose-effect and/or time-effect relationship.IL-6, IL-8 keep higher level in the inflammation Acute reaction phase, in atrophic gastritis shape
Cheng Hou, content reduce.IL-6, IL-8 can cause grain by the inflammatory cell based on induction neutrophil leucocyte in localized clusters
Cellular respiration is broken out, the neutrophil leucocyte for generating active oxygen (ROS), causing tissue inflammation [13], and activate can secrete IL-8,
The cell factors such as IL-1, IL-6, TNF-α constitute complicated cellular network, common to participate in stomach lining inflammation damnification.Continue repeatedly
Damage and the epithelial cell of reparation be susceptible to the variation of structure and function, the destruction and mutation of DNA, cause cell Proliferation with
The adjusting of apoptosis is disorderly.It is substantially reduced using rat blood serum IL-6, IL-8 level after qi and activate blood circulation high and low dose intervention, to
Mitigate its damage to gastric mucosa tissue.
(4) NF- κ B p65/I κ B are expressed and influence of the Blood rheology to it in PLGC rat gastric tissues
Many free radicals are generated in PLGC forming processes, biological effect, can also be by swashing other than cytotoxin acts on
Various transcription factors living are expressed to transcriptional activation target gene, participate in the proliferation of cell, apoptosis, differentiation, and inflammatory reaction is immunized
The processes such as reaction.NF- κ B are a kind of protein with transcriptional activation function, most common to be made of P65, P50 subunit, in endochylema
It with inhibit protein I κ B is combined to be formed tripolymer be in inactive state, by cell factor (such as TNF-α), endotoxin, oxygen oneself
When by stimulations such as bases, I κ B are easily degraded by proteases, NF- κ B, that is, quilt by protein kinase (ser kinases or other kinases) phosphorylation
Activation, is combined into nucleus with corresponding target gene, starts cell factors and other genes such as TNF-α, IL-6, IL-8
Transcription leads to a large amount of generations of inflammatory mediator, grade chain reaction is generated, to make inflammation continue and amplify [14-15].NF- κ B live
Existing apoptosis-promoting effect after change, and have Anti-G value.One side NF- κ B signal pathway activateds can raise pro apoptotic protein such as Fas, c-
The expression of myc, p53, IkappaB etc. [16-18], and then promote Apoptosis；On the other hand, after NF- kB activations can also on
Adjust expression such as TNF receptor associated factor (TRAF) TRAF1 and TRAF2, the apoptosis protein mortifier c- of suppression apoptotic proteins
IAP and c-IAP2, Fas related mortality domain protein sample interleukin converting enzyme inhibit albumen, Bcl-XL, A20 etc., and then press down
Apoptosis processed.The activation of NF- κ B is adjusting tumour cell existence, is inhibiting to play very crucial effect in apoptotic process.It grinds
To study carefully and shows [20-21], positive expression rates of the NF- κ B in normal structure, atrophic gastritis, stomach organization gradually rises, namely
When atrophic gastritis, the expression enhancing of NF- κ B in gastric epithelial cells, with the development of the state of an illness, the expression of NF- κ B increases
Abnormal cell proliferation caused by strong simultaneously gradually occupies leading position, can finally lead to the generation of gastric cancer.Therefore, NF- κ B are inhibited to live
It will be a feasible approach that changing, which reduces gastric cancer incidence,.
This studies have shown that model group rats gastric tissue NF- κ B, I κ B α are relatively normal, and group is significantly raised, in addition in endochylema
Expression is outer, also a large amount of nuclear expression, and NF- κ Bp65/I κ B α is prompted to participate in the generation of PLGC, development.Using Blood rheology and
After stomach answers spring intervention, high dose group, stomach are answered NF- κ B expression in spring group gastric mucosa and are substantially reduced, and I κ B alpha expressions are significantly raised,
NF- κ B p65 absorbances and positive area rate are significantly lower than model group.Prompt qi and activate blood circulation ruling by law may be by lowering NF- κ B
P65 express, up-regulation I κ B α come inhibit inflammatory cytokine (such as IL-2, TNF-α) transcription, release, it is anti-to mitigate mucosal inflammation
It answers.Blood rheology has the effects that promoting blood circulation and removing blood stasis, can improve stomach lining microcirculation, anti-oxidation stress reduces ROS and TNF-α
Etc. stimulation of the cell factors to stomach lining, to inhibit the activation of NF- κ B, and be allowed to can not with the specific bindings such as TNF-α,
The function of controlling gene transcription is suppressed inflammatory reaction that is related, therefore mitigating stomach lining.
(5) expression and Blood rheology influence to it of cyclinD1, p16 albumen in PLGC rat gastric tissues
The formation of gastric cancer is mainly manifested in from the formation of tumour in terms of biology angle：One side Apoptosis obstacle, separately
One side cell infinite multiplication, and with dysdifferentiation.Cell Proliferation is accelerated, and the cell of mutation cannot be withered again by cell
Mechanism removing is died, tumour is caused to be formed.And the basic vital movement such as cell Proliferation, differentiation, apoptosis and Association with Cell Cycle are close,
Therefore it sees in this sense, tumour is the disease of a kind of cell cycle exception, and the cell cycle is that cell Proliferation is total to what is broken up
Same access.
Cell sequentially completes its proliferation in the cell cycle by G1-S-G2-M.Wherein G1 phase test points are particularly important,
Cell is integrated and is transmitted to complicated intraor extracellular signal in this, decides whether to go successively to proliferating cycle, by this
After point, cell, which just complies with independent procedure independent of extracellular adjusting and controlling growth signal and postpones, completes whole cycle.Therefore, the G1 time limits
System point is the key point for controlling cell growth, determining cell fate.This adjusting function exception not only makes cell exogenous tune
It is blunt and be in uncontrolled proliferation to save signal reaction, will also be unable to retardance and have the DNA of damage to enter the S phases to replicate, and then generation is specific
Tumor phenotypes.Cyclin D_1 gene (cyclinD1) is the shaping regulatory factor that the G1 phases enter the S phases in the cell cycle, is also recognized
To be oncogene.The cyclinD1 assignments of genes gene mapping, can be by combining simultaneously active cell cyclin dependent protein kinase 4/ in 11q13
The activity of 6 (CDK4/6), Phospho-Rb make cell pass through the monitoring points G1/S and enter the S phases, complete the duplication of DNA, accelerate cell
Proliferation.The overexpression of cyclinD1 can shorten the time of G1 phases or accelerate G1 phase processes, it might even be possible to cell be made to flee from detection
The detection of point is directly entered the S phases, and uncontrolled cellular is caused to grow.CyclinD1 is the key protein of G1 phase cell proliferation signals.It grinds
Study carefully find cyclinD1 in gastric cancer there are gene magnification and protein overexpression, but, this prompt unrelated with tumor differentiation degree
CyclinD1 abnormal expressions may be the molecular events of early stage during gastric cancer occurrence and development.
P16 tumor suppressor genes are also known as CDK41 genes, are first and are found to directly act on the cell cycle, inhibit cell point
The tumor suppressor gene split is a kind of cyclin dependant inhibiting factor (CDKI), it by inhibit cyclin according to
Negative regulation function from the activity on cell period of bad property kinases (CDKs), is positioned at 9p21, in regulation of cell proliferation, p16 bases
Because being that the effect of cell cycle is inhibited mainly to be realized by cyclinS-CDK4,6-pRb-E2F approach.P16 albumen with
CyclinD1 is competed and the combination of CDK4/6, and the protein kinase activity of cyclinD1/CDK4/6 compounds is made to lose, with regulation and control
G1-S phase stuck points, to prevent, control cell division, Cell differentiation inducing activity plays negativity adjustment effect, common to complete to cell
The regulation and control of proliferating cycle.But when p16 protein delations, mutation and when causing the improper expression of p16 albumen, cell Proliferation will be out of control, leads
Cause tumour.It is significantly lower than normal gastric mucosa and non-cancerous issue in expression of p 16 in gastric cancer positive rate.
Therefore, P16 is negative regulatory factor, and cyclinD1 is positive regulatory factor；Positive negative regulatory factor interaction, remains
Dynamic equilibrium influences the process of G1 phases by adjusting the functional status of Rb albumen.Many studies have shown that the activation of NF- κ B accesses
It can promote the growth of gastric cancer and other tumours, NF- κ B that can adjust gastric cancer by combining the promoter of cyclinDs to adjust its expression
Growth.
The study find that normal rats liver cell has a small amount of cyclinD1 and p16 positive expressions, with cytoplasm and cell
Nuclear expression, model group rats hepatic tissue have cyclinD1 and p16 high expression, and positive expression is in nucleus, more normal group
Apparent increase, difference have very significant (P ＜ 0.01, P ＜ 0.05).Show that cyclinD1 and p16 take part in the development of PLGC
Process, it may be possible to the earliest events that gastric cancer occurs.After answering spring intervention using Blood rheology and stomach, high dose group and stomach answer spring group
The cyclinD1 expression of gastric mucosa tissue is remarkably decreased, and qi and activate blood circulation high dose group p16 increases notable.Show Blood rheology
NF- κ B signal accesses may be activated to inhibit cyclinD1, up-regulation p16 expression prevention gastric precancerous lesions by reducing pro-inflammatory cytokine.
But signal transduction pathway is a complicated network, influences each other, is associated between each channel, there is
Phosphorylation and dephosphorylation balance promote apoptosis and inhibit the balance regulation mechanism of apoptosis, once these balances or regulatory mechanism quilt
Break, it is possible to lead to pathologic process.The mechanism of each bars Signal Transduction Pathways and these Pathway Activations will be that PLGC is studied
Emphasis and hot spot.
The present invention improves the degree of inflammation and atrophy degree of gastric tissue, mitigates mucosa tissue hyper-proliferative；Adjust cell
Cytokine network improves Serum TNF-α activity, reduces IL-6, IL-8 generation；Inhibit gastric tissue NF- κ B/I κ B alpha signal conduction paths
Middle key protein NF- κ B expression and activation；Inhibit the expression 1 of gastric tissue cyclinD, up-regulation p16 expression prevention gastric precancerous lesions.
Claims (2)
1. a kind of compound stomach medicine of qi and activate blood circulation, characterized in that by Radix Codonopsis 20g, Radix Astragali 30g, fructus amomi 9g, Radix Notoginseng 9g, RADIX ANGELICAE SINEINSIS
20g, fingered citron 10g, Rhizoma Chuanxiong 15g, curcuma zedoary 12g compositions, 125 grams of meter/patch 200 paste totally.
2. a kind of preparation method of the compound stomach medicine of qi and activate blood circulation as described in claim 1, characterized in that by above-mentioned medicinal material,
It soaks 0.5 hour, decocts 2 times, for the first time plus 10 times are measured water, boil extraction 1.5 hours；Second plus 8 times of amount water, boil
Extraction 1.0 hours is concentrated into 6000 milliliters after merging extracting solution twice, and filling to 500 mL of saline bottles, scarfing sterilizes, refrigeration
It is i.e. spare.
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