CN103845695A - Compound stomach medicine for benefiting qi for activating blood circulation and preparation method thereof - Google Patents
Compound stomach medicine for benefiting qi for activating blood circulation and preparation method thereof Download PDFInfo
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Abstract
The invention provides a compound stomach medicine for benefiting qi for activating blood circulation. The stomach medicine comprises the following components by weight: 20g of codonopsis pilosula, 30g of astragalus membranaceus, 9g of fructus amomi, 9g of pseudo-ginseng, 20g of Chinese angelica, 10g of fingered citron, 15g of ligusticum wallichii, and 12g of curcuma zedoary, wherein each patch is 125g, and totally 200 patches are provided. The preparation method comprises the following steps: soaking the medicinal materials in water for 0.5 hour; decocting twice: adding 10 times water for the first time and boiling and extracting for 1.5 hours, and adding 8 times water for the second time and boiling and extracting for 1.0 hour; and combining two extracts, concentrating to 6,000ml, filling into a 500ml saline bottle, cutting, disinfecting and refrigerating for later use. According to the compound stomach medicine, the inflammation degree and atrophy degree of gastric tissues can be improved, over-proliferation of gastric mucosa can be reduced, cytokine network can be regulated, the serum TNF-alpha (tumor necrosis factor) activity can be improved, generation of IL-6 (interleukin-6) and IL-8 (interleukin-8) can be reduced, expression and activation of critical protein NF-kappa B (nuclear factor-kappa B) in a gastric tissue NF-kappa B/I kappa B alpha signal conduction path can be inhibited, expression 1 of gastric tissue cyclin D is inhibited, and p16 is up-regulated to prevent and treat precancerous lesions of gastric carcinoma.
Description
Technical field
The present invention relates to a kind of Chinese medicine and preparation method thereof, be specifically related to stomach medicine of a kind of benefiting QI for activating blood circulation and preparation method thereof.
Background technology
China is the High Risk For Gastric Cancer area, has every year 15～160,000 people to die from gastric cancer, and its case fatality rate occupy the prostatitis of various malignant tumor.Early diagnosis and early treatment are the only effective measures that improve at present gastric cancer survival rate, and the early diagnosis of gastric cancer mainly relies on the monitoring of gastric cancer variation in early stage with anti-.Gastric cancer variation in early stage comprises precancerous condition and precancerous lesion, and Gastric precancerous condition is clinical concept, refers to gastric precancer diseases, and modal is atrophic gastritis; Gastric precancerous lesion (Precancerous lesions of gastric cancer, PLGC) be pathological concept, comprise the life of gastric epithelial intestinalization and dysplasia, chronic atrophic gastritis occurs together, and epithelium intestinalization is given birth to or dysplasia is the high risk factor that gastric cancer occurs.Research shows, gives birth to from normal gastric mucosa-atrophic gastritis-intestinalization, and apoptosis and hypertrophy are proportionate; And from intestinalization life-dysplasia-gastric cancer, apoptosis and hypertrophy are negative correlation, the preliminary stage that prompting occurs in gastric cancer, gastric epithelial cell group is extremely unstable, exists the unbalance of apoptosis and hypertrophy.Therefore, inquiring into the pathogenesis of gastric precancerous lesion, effectively prevent and treat it and development occurs be of great significance reducing gastric cancer incidence rate, how to prevent and treat in early days and reverse PLGC and reduce the sickness rate of gastric cancer, is the focus of research at present.
Cyclin D1 (cyclinD1) is the shaping regulatory factor that in cell cycle, the G1 phase enters the S phase, is the key protein of G1 phase cell proliferation signal, is considered to oncogene.Research finds that cyclinD1 exists gene amplification and protein overexpression in gastric cancer, but irrelevant with Tumor Differentiation degree, and this prompting cyclinD1 abnormal expression may be early stage molecular events in gastric cancer generation evolution.
P16 is that first is found to directly act on cell cycle, suppresses fissional antioncogene.In regulation of cell proliferation, p16 albumen is combined with CDK4/6 with cyclinD1 competition, stops, controls cell division, Cell differentiation inducing activity, performance negativity regulating action.But while causing the improper expression of p16 albumen when p16 protein delation, sudden change, cell proliferation, by out of control, causes tumor to occur.Be starkly lower than normal gastric mucosa and non-cancerous issue at expression of p 16 in gastric cancer positive rate.
Nuclear Factor-Kappa B (NF-κ B) is the important factor that controlling gene is transcribed, and Recent study shows that it plays a significant role in generation, development and the apoptotic regulation and control of tumor.There is two-ways regulation relation in it and apoptosis, the transcriptional expression of numerous albumen that have anti-apoptotic effect is had to the effect that induction is raised, and regulates by the unbalanced expression to apoptotic proteins and anti-apoptotic proteins, and Cell protection is avoided apoptosis and threatened.The anti-apoptosis function of NF-κ B can also by and tumor suppressor protein (as p53 etc.) between direct interaction be achieved.Many tests confirm the activation that NF-κ B expresses in stomach cancer cell and tissue, closely related with formation, development, infiltration, the transfer of gastric cancer, and are determining to a certain extent patient's prognosis.But whether also have NF-κ B high expressed at PLGC, the relation between itself and cyclinD1, p16 how on earth, all still not fully aware of at present.
PLGC belong to motherland's medical science " gastric abscess ", " feeling of fullness ", " noisy " and wait category, take taste deficiency of both QI and YIN as this, or the double stagnation of QI, blood stasis, the comprehensive syndrome of simulataneous insufficiency and excessive.From " prolonged illness must be empty, pathogen usually intruding into collateral in protracted disease " theory of Chinese medical science analysis, in conjunction with its medical history, pathology reflection, we think that weakness of the spleen and stomach is its main pathogenesis, and obstruction of collaterals by blood stasis is its pathology key.Chinese medicine and Chinese medicine compound rest on the clinical observation stage to the research of PLGC more at present, to the experimentation deficiency of Mechanism of TCM, make treatment by Chinese herbs lack reliable objectively theoretical foundation, lack further screening and the checking of effective formula and drug.Yiqi Huoxue Recipe is mainly made up of Radix Codonopsis, the Radix Astragali, Fructus Amomi, Radix Notoginseng, Radix Angelicae Sinensis, Fructus Citri Sarcodactylis, Rhizoma Curcumae.For applying the agreement side of more than 10 years in institute, definite to chronic atrophic gastritis companion intestinal clinical efficacy, this research is point of penetration from NF-κ B, cyclinD1 and p16 pass, inquires into benefiting QI for activating blood circulation method to the PLGC mechanism of action, gets involved in early days PLGC treatment theory and experimental basis are provided for Chinese medicine.
Summary of the invention
For the deficiencies in the prior art, the invention provides compound stomach medicine of a kind of benefiting QI for activating blood circulation and preparation method thereof, improve degree of inflammation and the atrophy degree of gastric tissue, alleviate mucosa tissue hyper-proliferative; Regulate cytokine network, improve TNF-α activity, reduce IL-6, IL-8 generation; Suppressing key protein NF-κ B in gastric tissue NF-κ B/I κ B alpha signal pathway expresses and activation; Suppress the expression 1 of gastric tissue cyclinD, raise p16 and express control gastric precancerous lesion.
The technical solution adopted in the present invention is: a kind of compound stomach medicine of benefiting QI for activating blood circulation, it comprises Radix Codonopsis 20g, Radix Astragali 30g, Fructus Amomi 9g, Radix Notoginseng 9g, Radix Angelicae Sinensis 20g, Fructus Citri Sarcodactylis 10g, Rhizoma Chuanxiong 15g, Rhizoma Curcumae 12g, counts 125 grams/subsides, totally 200 pastes.
A compound stomach medicine for benefiting QI for activating blood circulation, its preparation method, by above-mentioned medical material, soaks 0.5 hour, decocts 2 times, adds for the first time 10 times of water gagings, boils and extracts 1.5 hours; Add for the second time 8 times of water gagings, boil and extract 1.0 hours.After merging extracted twice liquid, be concentrated into 6000 milliliters, fill to 500 mL of saline bottle, scarfing, sterilization, cold preservation is for subsequent use.
Compound stomach medicine of a kind of benefiting QI for activating blood circulation and preparation method thereof, middle dosage: 21g/kg/d, by the low middle Senior Three dosage group of ratio setting of 1: 2: 4, high dose group is: 42g/kg/d, general fluid extract is all prepared into high dose group, when administration, appropriateness dilution is used, press the dosage of 1mL/100g/d body weight, be 10mL/kg/d, the crude drug content of crude drug content=(42g/kg/d)/(10mL/kg/d)=4.2g/mL is high dose group fluid extract concentration, extractum amount=25000g/ (4.2g/mL)=5952mL, about 6000mL.
PLGC belongs to the categories such as the traditional Chinese medical science " gastric abscess ", " feeling of fullness ", and ancient Chinese medicine doctor is just recognized its pathogeny, clinical manifestation in early, and has proposed corresponding therapy and formula.PLGC is many to be attacked or does not relax because of diet internal injury, feelings will because of exopathogen, causes incoordination between the liver and stomach, gastric qi to become estranged, lead to fall in dereliction of duty, clear YANG failing to ascend, turbid pathogen and stop, with the passing of time dysfunction of the spleen in transportation, and it is moist that congestion resistance network causes body of stomach to lose, gastric gland atrophy.
" Treatise on the spleen and stomach " said: " taste deficiency is all disorders of blood ".Sun Weifeng thinks that the insufficiency of the spleen stasis of blood that causes is mainly by three aspects: the one, and transformation failure of spleen causes the stasis of blood: the 2nd, change source scarcity causes the stasis of blood; The 3rd, failure of the spleen to keep the blood flowing within the vessels causes the stasis of blood.Weng Weiliang thinks, disease, in the regular period, develops into certain phase, will affect the operation of QI and blood, cause disorder of QI and blood, solidifying, the dampness of the deficiency of vital energy, the stagnation of QI, QI rising in reverse order, the deficiency of YIN, cold coagulation, accumulation of heat, expectorant, body fluid deficiency liquid are few etc. generally all can the hyperamization stasis of blood, thus " all kinds of diseases and ailments are the stasis of blood all ".Dan Zhaowei points out that the pathogenesis key of CAG is deficiency of spleen-QI and stomach-QI, qi depression to blood stasis, and diet is careless or disloyal, or is hindered by feelings will, undermines taste, with the passing of time occur deficiency of spleen-QI and stomach-QI, heresy enters blood network, stomach network stasis blocking, insufficiency of vital energy and blood, cause gastric mucosa atrophy, pathogenesis key is at deficiency of spleen-QI and stomach-QI, blood-stasis internal-depression.
PLGC is take weakness of the spleen and stomach as this, and taste consumption wound is root, and the stasis of blood is stagnant does not go, and compound is not raw, and malicious heresy is gradually dark, evil poly-for a long time very or void or poison or the stasis of blood or many persons be mingled with, and so forms vicious cycle, and with the passing of time stomach is become homeless foster, mucosa attenuation, body of gland atrophy, becomes primary disease eventually, forms the gesture of heap soil or fertilizer over and around the roots in " void ", " stasis of blood "." stasis of blood " meets " two-hit theory " it is new paathogenic factor, can be used as again the performance of precancerous lesion.According to " deficiency syndrome should be treated by tonifying method; actually loose it ", the theory of " prolonged illness must be empty, pathogen usually intruding into collateral in protracted disease ", in view of primary disease etiology and pathogenesis it " void ", " stasis of blood ", be the rule for the treatment of therefore draft benefiting QI for activating blood circulation method, side is made up of Radix Codonopsis, the Radix Astragali, Radix Notoginseng, Radix Angelicae Sinensis, Fructus Amomi, Fructus Citri Sarcodactylis, Rhizoma Curcumae.Concrete side separates: ginseng, stilbene are monarch drug altogether, invigorating the spleen and replenishing QI, spleen-benefiting mind-tranquilizing, and mutual reinforcement between is for using; Ministerial drug is made up of Radix Notoginseng, Radix Angelicae Sinensis, and being longer than enriches blood invigorates blood circulation; Adjuvant drug: Fructus Amomi, Fructus Citri Sarcodactylis, Rhizoma Curcumae.
The Radix Astragali: sweet in the mouth; Slightly warm in nature; Return lung, spleen channel.Have tonifying Qi and lifting yang, benefit and defend the effects such as consolidating superficial resistance, promoting pus discharge and tissue regeneration strengthening be swollen, " book on Chinese herbal medicine meets former " calls its " can all void of the five internal organs ", is the key medicine of enriching spleen-QI.
Radix Codonopsis: sweet in the mouth is flat; Return spleen, lung meridian.Merit is being mended strengthening spleen and lung, supplementing QI for promoting the production of body fluid." book on Chinese herbal medicine is new again ": " invigorating the spleen and replenishing 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 is made a concerted effort so that " the vigorous spleen being able to resist against invasion of pathogen ", and with Zhi Qiben, righting is to get rid of evils.
Radix Notoginseng sweet in the mouth, micro-hardship, warm in nature, return liver, stomach warp, be classical blood circulation promoting and blood stasis dispelling good medicine, both kind removing stasis to stop bleeding, subduing swelling and relieving pain, again can Xu-tonic; " book on Chinese herbal medicine is looked for the truth " cloud: " the bitter temperature of Radix Notoginseng abnormal smells from the patient, can with its blood stasis of blood systemization.”
Radix Angelicae Sinensis: sweet in the mouth, pungent, micro-hardship; Return liver, the heart, spleen channel.Having enriches blood invigorates blood circulation, menstruction regulating and pain relieving, effect of loosening bowel to relieve constipation." book on Chinese herbal medicine through hundred kinds of records " cloud: " when the medicine that is classified as blood man and must uses ... the real product of wanting for nourishing blood.”
Fructus Amomi is pungent, warm, returns spleen, stomach, kidney channel.The turbid circulation of qi promoting of effectization, warming middle-JIAO.In " property of medicine opinion ", say: " main cold air stomachache ... the digesting and absorbing the essence of foodstuff that disappears, warm taste." be the tune stomach key medicine of being amusing.
Fructus Citri Sarcodactylis acrid, bitter, warm, returns spleen, stomach, liver, lung meridian, effect regulating the flow of Qi and the middle Jiao, and " book on Chinese herbal medicine is just read " said: " Fructus Citri Sarcodactylis, deficiency of YIN-blood person, also dislikes its dry ear.
Rhizoma Curcumae: acrid in the mouth, hardship; Warm in nature; Return liver, spleen channel.The merit of existing circulation of qi promoting removing blood stasis, removing food stagnancy pain relieving, also can dredge gas and QI invigorating, when empty people is by should the hold concurrently merit of tonification of Rhizoma Curcumae.Essentials of Matea Medica is sayed it, and " addiction that disappears is stimulated the menstrual flow, appetizing relieving dyspepsia, removing toxic substances pain relieving." " Bencao Jingshu " cloud: " and taste element is weak and without stagnant person, the anti-energy loss vital qi of using, food is not more disappeared and a little less than taste benefit ".Visible, Rhizoma Curcumae is applicable to the pathological state of this patient take void as this simulataneous insufficiency and excessive especially, and the Radix Astragali obtains Rhizoma Curcumae QI invigorating and in not stopping up, Rhizoma Curcumae obtains the Radix Astragali, breaks through and does not just hinder.Interior abnormal fermentation, dispels gastrointestinal pneumatosis, and has good fragrant digestion promoting function, and can preventing or arresting vomiting.
Modern pharmacology is thought: the Radix Astragali can promote humoral immunization, cellular immunization, and enhancing body anoxia enduring and stress ability significantly reduce rabbit blood Hemorheological Indexes, and its character is identical with intensity and Radix Salviae Miltiorrhizae Injection; In addition; the Radix Astragali also has the effect of protection gastric mucosa; the Radix Astragali, Radix Angelicae Sinensis injection are carried out to Zusanli point injection; can obviously dwindle intercellular substance; recover epithelial cell and connect composite construction and effectively reverse intestinal and the dysplasia of mucosa, there is the effect, the Radix Astragali that suppress experimental chronic atrophic gastritis and also have antibacterial and suppress viral effect, cancer suppressing action.Radix Codonopsis energy Gastrointestinal motility adjustment, antiulcer, enhancing immunologic function, have impact to excited and two kinds of nervous process of inhibition.Radix Angelicae Sinensis there is antithrombotic and improve blood theomorphism, improve blood circulation, the effect such as enhancing human body immunity, anti-inflammatory and antalgic, removing oxygen-derived free radicals, anti peroxidation of lipid.Radix Notoginseng has hemopoietic function, can improve humoral immune function,, there is analgesic and anti-inflammatory effects, the atrophic lesion of gastric mucosa can be obviously treated, and epithelioglandular atypical hyperplasia and intestinal epithelial metaplasia can be reversed, there is prophylaxis of tumours effect.Fructus Amomi can strengthen the function of stomach, and the secretion of facilitating digestion liquid, can promote intestinal peristalsis promoting, energy anticoagulant; Rhizoma Curcumae: modern pharmacological research confirmation, Oleum Curcumae has antitumor, antiplatelet aggregation, antimicrobial antiphlogistic, improves gastrointestinal smooth muscle motion, has physiologically active widely, especially effect uniqueness aspect inhibition tumor.Fructus Citri Sarcodactylis has obvious inhibitory action to intestinal smooth, can blood vessel dilating, too many levels immunologic function is had to obvious facilitation, and can promote Peritoneal Macrophage Phagocytosis.
Figure of description:
Fig. 1 is the impact of benefiting QI for activating blood circulation method of the present invention on PLGC rat blood serum TNF-alpha expression;
Fig. 2 is the impact that benefiting QI for activating blood circulation method of the present invention is expressed PLGC rat blood serum IL-6;
Fig. 3 is the impact that benefiting QI for activating blood circulation method of the present invention is expressed PLGC rat blood serum IL-8;
Fig. 4 is benefiting QI for activating blood circulation method of the present invention is organized NF-κ Bp65 protein expression impact on PLGC rat stomach;
Fig. 5 is benefiting QI for activating blood circulation method of the present invention is organized p16 protein expression impact on PLGC rat stomach;
Fig. 6 is benefiting QI for activating blood circulation method of the present invention is organized p16 protein expression impact on PLGC rat stomach.
The specific embodiment
Below in conjunction with the detailed embodiments of the present invention of embodiment:
A compound stomach medicine for benefiting QI for activating blood circulation, it comprises Radix Codonopsis 20g, Radix Astragali 30g, Fructus Amomi 9g, Radix Notoginseng 9g, Radix Angelicae Sinensis 20g, Fructus Citri Sarcodactylis 10g, Rhizoma Chuanxiong 15g, Rhizoma Curcumae 12g, counts 125 grams/subsides, totally 200 pastes.Above-mentioned medical material, soaks 0.5 hour, decocts 2 times, adds for the first time 10 times of water gagings, boils and extracts 1.5 hours; Add for the second time 8 times of water gagings, boil and extract 1.0 hours.After merging extracted twice liquid, be concentrated into 6000 milliliters, fill to 500 mL of saline bottle, scarfing, sterilization, cold preservation is for subsequent use.Assess this Chinese medicine effect below by SD rat.
(1) material
1. laboratory animal
64 of SD rats, clean level, male and female half and half, body weight 90-110g, Shanghai Xi Pu-Bi Kai laboratory animal company limited provides, and credit number: SCXK (Shanghai) 2008-0016, raises the Experimental Animal Center in Zhejiang University of Traditional Chinese Medicine.
2. experimental article
(1) experimental drug
MNNG (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, the Radix Astragali, Fructus Amomi, Radix Notoginseng, Radix Angelicae Sinensis, Fructus Citri Sarcodactylis, Rhizoma Chuanxiong, Rhizoma Curcumae) is made suspension by the Drug Manufacturing Room of Hangzhou Chinese Medicinal Hospital, and every ml is containing crude drug 3.1714g.
(2) main agents
Tumor necrosis factor-alpha (TNF-α) (lot number 11110403), interleukin-6 (IL-6) (lot number 11110404), interleukin-8 (IL-8) (lot number 11110405) is Shanghai Zhuo Kang biotech firm product.
Paraformaldehyde (lot number 20051208) Tianjin City Chemical Agent Research Institute product.
NF-κ Bp65 rabbit polyclonal antibody (lot number G2010) is U.S. NeoMarkers product, working concentration 1: 50.
I κ B-α rabbit anti-(lot number G1411) is U.S. Santa Cruz product, and working concentration is 1: 50.
P16 (lot number I0110) is U.S. Santa Cruz product, and working concentration is 1: 50.
Cyclind1 (lot number J1810) is U.S. Santa Cruz product, and working concentration is 1: 50.
Envision two step method SABC test kit (lot number 00029661) is Dako company of Denmark.
PBS phosphate buffer (ZLI-9062), solution final concentration: 0.01M, citrate buffer (ZLI-9065), solution final concentration: 0.01M, concentrated type DAB test kit (ZLI-9032) (lot number 60882801), by Beijing, Bioisystech Co., Ltd of Zhong Shan Golden Bridge produces.
DAB nitrite ion (lot number 60119) provides for Beijing Bioisystech Co., Ltd of Zhong Shan Golden Bridge.
(3) experimental apparatus, equipment
(2) method
The foundation of 1.PLGC rat model
Modeling group rat adopts add MNNG every day in rat drinking-water, and concentration is 167 μ g/mL, presses the MNNG solution of 0.017mol/L concentration simultaneously every afternoon, and 1ml/ Mus gavage attacked, and continuous 8 weeks, feeds with common rat feed.
2. animal grouping and administration
64 rats are divided into 56 of 8 of normal group and modeling groups at random, put to death at random 10 modeling group rats and cause gastric precancerous lesion model through proved by pathology through modeling intervention, residue modeling group rat is divided into 12 of model group, 12 of benefiting QI for activating blood circulation high dose group, 12 of benefiting QI for activating blood circulation low dose group, 10 of WEIFUCHUN groups at random.Benefiting QI for activating blood circulation high and low dose group is given respectively the Chinese medicine gavage treatment of 1.6g/kg body weight d and 0.8g/kg body weight d every day, WEIFUCHUN group is given the treatment of WEIFUCHUN suspension (1.23g/kg.d) gavage, model group, to wait normal saline gavage of capacity, is total to continuous 12 weeks.Experimental session is observed the ordinary circumstance of each group of rat.
3. collection of specimens
After last administration, process on an empty stomach rat morning next day, and 3% pentobarbital sodium anesthesia, gets gastric tissue, cuts off and makes gross examination of skeletal muscle along greater gastric curvature, get lesser curvature side hole body boundary gastric tissue, fix with 4% paraformaldehyde, step by step dehydration of alcohol, dimethylbenzene is transparent, paraffin embedding, and it is stand-by that routine is prepared paraffin section.
4. detect index and method
(1) histopathology changes: conventional H E dyeing and Masson trichrome stain, observe its gastric mucosa atrophy degree and thickness.
(2) TNF-α level: adopt ELISA method to measure, carry out in strict accordance with test kit operating procedure.
(3 blood serum IL-6s, IL-8 level: adopt measured by radioimmunoassay, carry out in strict accordance with test kit operating procedure.
(4) expression of NF-κ Bp65/I κ B α, cyclinD1, p16 albumen in mucosa tissue: adopt SABC Envision method to detect.Main operational steps is as follows: paraffin section is de-cured to water, PBS rinsing, and PBS rinsing after pressure cooker antigen retrieval, PBS rinsing after 3%H202 room temperature 10min, drips 37 ℃ of primary antibodies and hatches 60min, PBS rinsing, dropping two resists 37 ℃ and hatches 60min; Hatch 60min, PBS rinsing, DAB fully develops the color and stops and fully washing after 1～2min, and haematoxylin is redyed, dehydration mounting; Wherein replace primary antibodie as negative control using PBS.
5. diagnostic criteria
(1) gastric mucosa tissue pathological observation, with reference to the pathology standard in national chronic gastritis seminar National Consensus in 2000.
Chronic inflammatory disease: normal (0 point): the every high power field of mononuclearcell is no more than 5.Slightly (1 point): chronic inflammatory cells is less and be confined to mucosa shallow-layer, is no more than 1/3 of mucous layer.Moderate (2 points): chronic inflammatory cells comparatively dense, exceed 1/3 of mucous layer, reach 2/3.Severe (3 points): chronic inflammatory cells is intensive, occupies mucosa holostrome.
Atrophy: slight (1 point): intrinsic body of gland number minimizing is no more than 1/3 of original body of gland, and most of body of gland still retains.Moderate (2 points): intrinsic body of gland number minimizing exceedes 1/3, but does not exceed 2/3.The irregular distribution of remaining body of gland.Severe (3 points): intrinsic body of gland number minimizing exceedes 2/3, and only residual minority body of gland, disappears even completely.
Intestinal gland metaplasia: intestinal part account for body of gland and the superficial epithelium gross area below 1/3 for slight (1 point), 1/3～2/3 be moderate (2 points), more than 2/3 be severe (3 points).
Dysplasia: (1) low dysplasia: mostly occur at mucomembranous surface, nucleus close-packed arrays is in substrate, the intensive engrain of nuclear chromatin, core increases, and karyokinesis resembles less, and core is generally positioned at the half of cell or below 2P3, still keeps certain polarity.Gland structure is slightly irregular, and form is slightly disorderly, seldom occurs back-to-back and common wall.(2) high grade dysplasia: cell pleomorphism is obvious, visible pathological mitotic figure, core floats up to cell top, and arrangement disorder forms multiple stratification, partially or completely loses polarity.Nuclear hyperchromatism, kernel is large, is blister.Gland structure is abnormal, has back-to-back, altogether wall and the phenomenon of sprouting more.1 point of low atypical hyperplasia, 3 points of high grade dysplasias, low-2 points of high grade dysplasias.
(2) Masson dyeing criterion: collagen fiber are blue, and muscle, elastic fibers take on a red color, and cellulose is aubergine, and erythrocyte is Chinese red, and nucleus is blue brown.
(3) SABC detects cyclinD1, NF-κ Bp65, I κ B α, p16 result evaluation standard: ImmunohistochemistryResults Results is all observed under identical conditions, NF-κ Bp65, I κ B α positive cell show as endochylema/karyon and are brown yellow granule, p16 positive cell shows as karyon and is brown yellow granule, and cyclinD1 positive cell shows as karyon and is brown yellow granule.Result evaluation standard: positivity index: i.e. IOD/1228800 (IOD area calculates with pixel unit) or IOD/72041.227 (IOD area calculates with actual size μ m2 unit), the SABC section positive region developing the color with DAB is brown color, positivity index represents that total optical density all assigns to the value of this photo, it is the yellow degree of captured photo, be worth larger, photo is more yellow, evaluates the positive degree of the corresponding tissue of photo with this.
6. statistical analysis
Adopt SPSS10.5 software kit statistics, mean ± standard deviation for data
represent, adopt respectively one factor analysis of variance, non parametric tests etc. according to the character of data and research purpose.
Three, result
(1) rat ordinary circumstance
When experiment finishes, model group, benefiting QI for activating blood circulation low dose group respectively have 2 rats deaths; Benefiting QI for activating blood circulation high dose group and WEIFUCHUN group respectively have 1 rats death; Normal rats is without death.Rats death is mainly because the anti-esophagus suction pulmonary that flows into after gavage causes death.
Normal rats fur is closely next to the skin, color white light pool, and two have god, and movable quick, appetite is more, and stool is graininess, and tail color is light red.Rat model fur is fluffy and disorderly, matt, irritability, and appetite is less, and lethargy stool abnormal smells from the patient is smelly dirty, and how partially rare matter is or dry rare uncomfortable.Benefiting QI for activating blood circulation high and low dose group, WEIFUCHUN group rat also have fur fluffy and disorderly, and appetite reduces, irritability, but degree is all light compared with model group rat, and stool is still shaped.
Rats in normal control group gastric mucosa pinkiness, glossiness is bright-coloured, mucosal fold rule out of shape, surface is with mucus, flexible.Model group gastric mucosa is lead, and glossiness is dark and gloomy, and coat of the stomach compared with normal group is thin, and mucosal fold is smooth, and wherein 5 rat stomach are organized visible hemorrhage rotten to the corn point.Benefiting QI for activating blood circulation high and low dose and WEIFUCHUN group are all improved compared with model group, have no hemorrhage rotten to the corn point.
(2) gastric mucosa of rat histology changes
PLGC rat model gastric tissue is learned and is changed normal rats gastric mucosal cell marshalling under light microscopic, Glands morphology rule, and accidental inflammatory cell is invaded profit lamina propria.The intrinsic body of gland number of PLGC model group gastric mucosa of rat reduces, and the visible inflammatory cell of lamina propria is invaded profit, part visible cell arrangement disorder, and Glands morphology is irregular, and the visible slight enterocyteization of 4 example is raw, has no obvious dysplasia (seeing Fig. 1-3).
With normal group comparison, model group rat stomach tissue adherence thickness obviously reduces (P < 0.01); After Drug therapy with model group comparison, benefiting QI for activating blood circulation high dose group rat stomach tissue thickness obviously improves (P < 0.05), benefiting QI for activating blood circulation low dose group, WEIFUCHUN group rat stomach tissue thickness are slightly thick compared with model group, but no significant difference (P > 0.05), between three pharmaceutical intervention groups, benefiting QI for activating blood circulation high dose group rat stomach tissue thickness improves obviously (P < 0.05) compared with benefiting QI for activating blood circulation low dose group and WEIFUCHUN group.
With normal group comparison, model group rat stomach tissue inflammation degree obviously increases (P < 0.01); After Drug therapy, benefiting QI for activating blood circulation high and low dose group, WEIFUCHUN group rat stomach tissue inflammation score degree obviously alleviate (P < 0.05, P < 0.01) compared with model group; But between three pharmaceutical intervention groups and normal group and each medicine, comparing difference is without significance (P > 0.05).Normal rats has no atrophy and occurs, after Drug therapy with model group comparison, benefiting QI for activating blood circulation high dose group rat stomach Telatrophy score degree obviously alleviates (P < 0.05), benefiting QI for activating blood circulation low dose group, WEIFUCHUN group rat stomach Telatrophy score degree difference is without significance (P > 0.05), between three pharmaceutical intervention groups and normal group and each medicine comparing difference without significance (P > 0.05) in table 1.
The comparison of the each group of table 1. rat stomach tissue inflammation degree, atrophy degree score
* P < 0.05 compared with normal group, * * P < 0.01; Δ P < 0.05 compared with model group, Δ Δ P < 0.01; With benefiting QI for activating blood circulation high dose group than P < 0.05
(3) impact of benefiting QI for activating blood circulation method on PLGC rat blood serum TNF-α, IL-6, IL-8
Model group rat blood serum TNF-α, the horizontal compared with normal group of IL-6, IL-8 reduce, and wherein IL-8 level obviously reduces (P < 0.01); Benefiting QI for activating blood circulation high and low dose group and WEIFUCHUN group rat blood serum IL-6 level obviously reduce (P < 0.01 compared with model group, P < 0.05), no significant difference between three pharmaceutical intervention groups (P > 0.05); WEIFUCHUN group rat blood serum IL-8 level raises (P < 0.05) in table 2 compared with benefiting QI for activating blood circulation high and low dose group.
The impact of table 2. benefiting QI for activating blood circulation method on PLGC rat blood serum TNF-α, IL-6, IL-8
* P < 0.05 compared with normal group, * * P < 0.01; Δ P < 0.05 compared with model group, Δ Δ P < 0.01; With benefiting QI for activating blood circulation high dose group than P < 0.05; With benefiting QI for activating blood circulation low dose group than ■ P < 0.05
(4) benefiting QI for activating blood circulation method is organized the impact of NF-κ Bp65, I κ B α protein expression on PLGC rat stomach
Normal rats gastric tissue has a small amount of NF-κ Bp65 positive expression, and take cytoplasm as main, few nucleus is expressed, and cytoplasm has a small amount of I κ B α positive expression.Model group rat stomach is organized all NF-κ Bp65 high expressed, and positive expression is in cytoplasm and nucleus, and gastric tissue I κ B α has high expressed, and positive expression is in cytoplasm.Model group rat stomach organizes NF-κ Bp65, the equal compared with normal group of I κ B α obviously to raise, and difference has significance (P < 0.01); Benefiting QI for activating blood circulation high dose group, WEIFUCHUN group rat stomach are organized NF-κ Bp65 to express obviously to reduce (P < 0.05) compared with model group, and I κ B α protein expression is compared with model group obviously raise (P < 0.01); Between three medicine groups, NF-κ Bp65 protein expression group difference is without significance, and benefiting QI for activating blood circulation low dose group I κ B α protein expression obviously reduces (P < 0.01) compared with benefiting QI for activating blood circulation high dose group and WEIFUCHUN group.See Fig. 4-5, table 3,4.
Table 3. benefiting QI for activating blood circulation method is organized the impact of NF-κ Bp65 protein expression on PLGC rat stomach
* P < 0.05 compared with normal group, * * P < 0.01; Δ P < 0.05 compared with model group, Δ Δ P < 0.01; With benefiting QI for activating blood circulation high dose group than P < 0.05; With benefiting QI for activating blood circulation low dose group than ■ P < 0.05
Table 4. benefiting QI for activating blood circulation method is organized the impact of I κ B α protein expression on PLGC rat stomach
* P < 0.05 compared with normal group, * * P < 0.01; Δ P < 0.05 compared with model group, Δ Δ P < 0.01; With benefiting QI for activating blood circulation high dose group than P < 0.05, P < 0.01; With benefiting QI for activating blood circulation low dose group than ■ P < 0.05, ■ ■ P < 0.01
(5) benefiting QI for activating blood circulation method is organized the impact of cyclinD1, p16 protein expression on PLGC rat stomach
Normal rats gastric tissue has a small amount of cyclinD1 and p16 positive expression, and take karyon as main, model group rat stomach is organized all cyclinD1 and p16 high expressed.Model group rat stomach is organized cyclinD1 and p16 compared with normal group obviously raise (P < 0.01, P < 0.05); Benefiting QI for activating blood circulation high dose group and WEIFUCHUN group rat stomach organize cyclinD1 expression obviously to reduce (P < 0.01, P < 0.05) compared with model group; Benefiting QI for activating blood circulation high dose group rat stomach organizes p16 to express compared with model group obviously raise (P < 0.01); Between three medicine groups, cyclinD1, p16 protein expression group difference are without significance.In table 5,6 and accompanying drawing 4-5.
Table 5. benefiting QI for activating blood circulation method is organized the impact of Cyclind1 protein expression on PLGC rat stomach
* P < 0.05 compared with normal group, * * P < 0.01; Δ P < 0.05 compared with model group, Δ Δ P < 0.01; With benefiting QI for activating blood circulation high dose group than P < 0.05, P < 0.01; With benefiting QI for activating blood circulation low dose group than ■ P < 0.05, ■ ■ P < 0.01
Table 6. benefiting QI for activating blood circulation method is organized the impact of p16 protein expression on PLGC rat stomach
* P < 0.05 compared with normal group, * * P < 0.01; Δ P < 0.05 compared with model group, Δ Δ P < 0.01; With benefiting QI for activating blood circulation high dose group than P < 0.05, P < 0.01; With benefiting QI for activating blood circulation low dose group than ■ P < 0.05, ■ ■ P < 0.01
Gastric cancer is the malignant tumor very high at whole world M & M, and global nearly half gastric cancer occurs in China, is in recent years rejuvenation trend, and case fatality rate occupy various malignant tumor prostatitis.Early diagnosis, early treatment are the unique effective measures that improve at present gastric cancer survival rate, and early diagnosis mainly relies on the monitoring to gastric cancer variation in early stage.Therefore, inquire into gastric precancerous lesion pathogenesis, effectively prevent and treat it and develop, become the emphasis of gastric cancer study on prevention, and be of great significance reducing gastric cancer incidence rate.
(1) foundation of gastric precancerous lesion rat animal model
Depend on to a great extent reliability and the repeatability of animal model for the active drug of the pathogenetic research of gastric precancerous lesion, screening control, the current animal model of gastric precancerous lesion rat animal modeling mainly contain at present gastric mucosa repeatedly damage method, immunity modeling method, low dose of x-ray local irradiation method, in conjunction with carcinogenic comprehensive molding method, active immunity in conjunction with mucosal lesion method etc.
Wherein that current domestic scholars is selected maximum gastric precancerous lesion models in conjunction with carcinogenic comprehensive modeling method, the application MNNG such as Japanese Sug-imura in 1967 administration soluble in water, successfully bring out the adenocarcinoma of stomach of Wistar rat, luring in cancer process, gastric mucosal lesion has experienced the evolution from erosion, atrophy, regenerative proliferation, dysplasia to canceration, these pathological changes accord with the being seen variation of clinical CAG gastroscope substantially, have also illustrated that the atrophy of gastric mucosa and dysplasia are seemingly must have pathological changes in experimental animal models gastric cancer process.The animal model that complex method take MNNG (MNNG) as basic mutagenic agent is set up.After this, many scholars improve on the method basis, to copy the model that more meets mankind's gastric precancerous lesion.
This research list of references report combination research in earlier stage, select MNNG carcinogen, and in rat drinking-water, adding concentration is the MNNG of 167 μ g/mL, presses the MNNG solution of 0.017mol/L concentration simultaneously every afternoon, and 1ml/ Mus gavage attacked, after continuous 8 weeks.Histopathology is visible, and normal rats gastric mucosa thickness is normal, and epithelium is complete, epithelial cell and body of gland marshalling, rule, and the boundary of Glandular Epithelium and Gland pipe is clear; The attenuation of model group gastric mucosa of rat, intrinsic body of gland number reduces, and the visible inflammatory cell of lamina propria is invaded profit, part visible cell arrangement disorder, Glands morphology is irregular, shows to quote for a long time MNNG and can make gastric mucosa barrier be damaged, and causes erosion, atrophy and part intestinal.Although part visible cell arrangement disorder, Glands morphology is irregular, have no obvious dysplasia, can extending the modeling time cause dysplasia? repeatedly gavage easily causes the complication that intubate gavage is relevant, as rats death rate increases, how the cost that has increased experiment, reduce mortality rate? be worth our further further investigation from now on.
(2) impact of benefiting QI for activating blood circulation method on MNNG induction gastric precancerous lesion gastric mucosa degree of inflammation, atrophy degree
PLGC is take taste deficiency of both QI and YIN as this in motherland's medical science, or the double stagnation of QI, blood stasis, the comprehensive syndrome of simulataneous insufficiency and excessive.From " prolonged illness must be empty, pathogen usually intruding into collateral in protracted disease " theory of Chinese medical science analysis, in conjunction with its medical history, pathology reflection, we think that weakness of the spleen and stomach is its main pathogenesis, and obstruction of collaterals by blood stasis is its pathology key.Yiqi Huoxue Recipe is mainly made up of Radix Codonopsis, the Radix Astragali, Radix Notoginseng, Radix Angelicae Sinensis, Fructus Amomi, Fructus Citri Sarcodactylis, Rhizoma Curcumae.Radix Codonopsis, Radix Astragali invigorating the spleen and replenishing QI, control insufficiency of the spleen basis; Radix Notoginseng, Radix Angelicae Sinensis blood circulation promoting and enriching; During Fructus Amomi circulation of qi promoting is adjusted, stomach function regulating is amusing and is helped ginseng, stilbene invigorating the spleen and regulating the stomach; Fructus Citri Sarcodactylis regulating the flow of Qi and the middle Jiao; Rhizoma Curcumae circulation of qi promoting removing blood stasis removing food stagnancy.Full side is coordinated mutually, the effect of playing altogether Yiqi-Huoxue-Huayu.
Normal rats gastric mucosal cell marshalling under light microscopic is found in this research, Glands morphology rule, and accidental inflammatory cell is invaded profit lamina propria.The intrinsic body of gland number of PLGC model group gastric mucosa of rat reduces, and the visible inflammatory cell of lamina propria is invaded profit, part visible cell arrangement disorder, and Glands morphology is irregular, and the visible slight enterocyteization of 4 example is raw, has no obvious dysplasia.Model group rat stomach tissue adherence thickness and degree of inflammation compared with normal group obviously reduce; After Drug therapy, with model group comparison, benefiting QI for activating blood circulation high dose group rat stomach tissue thickness obviously improves.Model group rat stomach tissue inflammation degree compared with normal group obviously increases; After Drug therapy, benefiting QI for activating blood circulation high and low dose group, WEIFUCHUN group rat stomach tissue inflammation score degree obviously alleviate compared with model group.Normal rats has no atrophy and occurs, and after Drug therapy, with model group comparison, benefiting QI for activating blood circulation high dose group rat stomach Telatrophy score degree obviously alleviates.Show that benefiting QI for activating blood circulation method has inhibitory action to the hyper-proliferative of MNNG induction gastric precancerous lesion gastric mucosa cell, the histopathology that can partly reverse gastric mucosa changes, reduce the release of inflammatory cell, improve mucosa nutritive situation, alleviate gastric mucosa tissue degree of inflammation and atrophy degree, alleviate gastric mucosa tissue damage.
(3) TNF-α, IL-6, IL-8 level and the impact of benefiting QI for activating blood circulation method on it in PLGC rat blood serum
Pathogenesis about PLGC is also indefinite, thinks at present and infection and immune relevant more.May relate to the pathological process of PLGC as TNF-α, the IL-6 of important inflammation and the immune-mediated factor and IL-8.TNF-α is a kind of important medium between specific immune response and inflammatory reaction, generation, evolution in inflammation play an important role, Chemokines and activity factor, appropriate TNF-α can participate in the activation inflammatory reaction of immunocyte, has infection, antiviral and antitumor action; Excessive TNF-α strengthens wall of micrangium permeability, produce platelet activating factor and chemotactic cytokine IL-8 by raising Some Adhesion Molecules on Endothelial Cells inducing endothelial cell, change mucosa vascular endothelial cell reactivity, cause blood capillary intravascular coagulation, cause cytoclasis, lysosome spills, inflammatory mediator discharges, and causes the damage of gastric mucosa endotheliocyte, makes Oligemia, and Gastric Mucosa Blood Flow is most important in the defense reaction of gastric mucosal barrier, thereby cause mucosal lesion.Meanwhile, it can also chemotactic neutrophilic granulocyte, strengthens it and engulfs kill capability, strengthens inflammation regional nodes cellular infiltration and propagation, changes thereby brought out a series of inflammation, increases the weight of mucosal inflammation reaction [8].TNF-α can stimulate body to produce a series of inflammatory cytokine (IL-1, IL-6 and IL-8) generation " water fall effect ", can cell death inducing, TNF-a can be under IL-6 participates in addition, induction thrombin forms, make endothelial cell surface become short coagulation from anticoagulation state, the little blood vessel of mucosa forms microthrombus on inflammation basis, cause mucosa microcirculation disturbance, the barrier function of impair gastrointestinal mucosa, can activate again Nuclear Factor kappa B etc. simultaneously, the latter has promotion cell survival, propagation and differentiation, anti-apoptosis activity, especially in, in the pathological change of severe atrophic gastritis, may exist the immunologic injury due to TNF-α, its expression in vivo also affects the generation of tumor, development.
IL-6 is mainly secreted by monocytes/macrophages, in the time of inflammation and stress, IL-6 can induce body to produce various acute phase albumen, it is the synthetic important medium of acute phase reactant, simultaneously, also be the important cytokine of Organism immunoregulation, participate in T, the propagation of B cell, the various biological effect of differentiation and NK cell and macrophage, the report that relevant IL-6 affects cancerous cell is not identical, the IL-6 that tumor cell generates may be served as a kind of autocrine or paracrine somatomedin, the membrane receptor effect special with cell surface, to the growth of tumor cell, the simultaneous phenomenon of tumor patient, the pathological process of disease and the order of severity of the state of an illness all have impact.
IL-8 is mainly subject to lipopolysaccharide or tumor necrosis factor (TNF-α), IL-1 etc. by monocytes/macrophages, histiocyte (endotheliocyte, fibroblast) etc. stimulates generation, is a kind of main chemotactic factor.Its centering granulocyte, basophilic granulocyte and T cell have chemotaxis, and can promote the activation of neutrophilic granulocyte, make neutrophilic granulocyte assemble and discharge superoxides and lysosomal enzyme gastric mucosa is local, its these biologic activity play a significant role in opposing pathogen process, but have also caused the damage [11] of mucosa tissue simultaneously.Research is found to express in the antrum infecting without Hp and obviously increase in antrum that IL-8 infects at Hp, and IL-8 level increasing the weight of and increase [12] with degree of inflammation.
This research is found, model group rat blood serum TNF-α, the horizontal compared with normal group reduction of IL-6, IL-8, and wherein IL-8 level obviously reduces (P < 0.01); Benefiting QI for activating blood circulation high and low dose group and WEIFUCHUN group rat blood serum IL-6 level obviously reduce (P < 0.01 compared with model group, P < 0.05), no significant difference between three pharmaceutical intervention groups (P > 0.05).Showing the generation of PLGC and transform may to lack the gastric mucosa nutrition scarcity, epithelial repair obstacle and the immunity of organisms that cause lowly relevant with TNF-α.After the treatment of benefiting QI for activating blood circulation method, gastric mucosa inflammation and atrophy degree obviously alleviate, and TNF-α level increases, and prompting impels TNF-α secretion to increase, may be its proliferative disorder that reverses PLGC gastric mucosa, thereby block one of its mechanism of action transforming to gastric cancer.But the effect of TNF-α is not adjusted to yet and approaches normal level, point out this research may not yet reach best dose-effect and/or time-effect relationship to the application of this medicine.IL-6, IL-8 all keep higher level at inflammation Acute response stage, and after atrophic gastritis forms, its content reduces.IL-6, IL-8 can be that main inflammatory cell is assembled in part by induction neutrophilic granulocyte, cause granulocyte respiratory burst, generate active oxygen (ROS), cause tissue inflammation [13], and the neutrophilic granulocyte of activation can be secreted the cytokines such as IL-8, IL-1, IL-6, TNF-α, form complicated cellular network, jointly participate in the damage of gastric mucosa inflammation.Easily there is the variation of 26S Proteasome Structure and Function in the epithelial cell of lasting injuring and repairing repeatedly, and the destruction of DNA and sudden change cause the adjusting disorder of Cell apoptosis and proliferation.After the intervention of application benefiting QI for activating blood circulation high and low dose, rat blood serum IL-6, IL-8 level obviously reduce, thereby alleviate its infringement to gastric mucosa tissue.
(4) NF-κ B p65/I κ B expresses and the impact of benefiting QI for activating blood circulation method on it in PLGC rat stomach tissue
In PLGC forming process, produce a lot of free radicals, its biological effect, except cytotoxin effect, is expressed thereby also can pass through to activate various transcription factor transcriptional activation target genes, participates in propagation, apoptosis, the differentiation of cell, inflammatory reaction, the processes such as immunoreation.NF-κ B is a kind of protein with transcriptional activation function, the most common by P65, P50 subunit forms, in endochylema it be combined with inhibitory κBα form trimer be inactive state, be subject to cytokine (as TNF-α), endotoxin, when oxygen-derived free radicals etc. stimulate, I κ B is by protein kinase (ser kinases or other kinases) phosphorylation and by proteasome degradation, NF-κ B is activated, entering nucleus combines with corresponding target gene, start TNF-α, IL-6, the cytokines such as IL-8 and other genetic transcriptions, cause a large amount of generations of inflammatory mediator, produce level chain reaction, thereby inflammation is continued and amplify [14-15].Existing apoptosis-promoting effect after NF-kB activation, has again anti-apoptotic effect.NF-κ B signal pathway activated can raise pro apoptotic protein as the expression of [16-18] such as Fas, c-myc, p53, IkappaB on the one hand, and then promotes apoptosis; On the other hand, after NF-kB activation, can also raise the expression that presses down apoptotic proteins as TNF (tumor necrosis factor) receptor associated factor TRAF1 and TRAF2, apoptosis protein mortifier c-IAP and c-IAP2, Fas dependency death domain protein sample interleukin converting enzyme Profilin, Bcl-XL, A20 etc., and then inhibited apoptosis.The activation of NF-κ B is played very crucial effect in regulate tumor cell existence, apoptosis inhibit process.Research shows [20-21], the positive expression rate of NF-κ B in normal structure, atrophic gastritis, stomach organization raises gradually, also while being chronic atrophic gastritis, in gastric epithelial cells, the expression of NF-κ B strengthens, along with the development of the state of an illness, the expression of NF-κ B strengthens the abnormal cell proliferation that causes dominate gradually, finally can cause the generation of gastric cancer.Therefore, suppress NF-kB activation reduction gastric cancer incidence rate and will be a feasible approach.
Originally studies show that, model group rat stomach organizes NF-κ B, the equal compared with normal group of I κ B α obviously to raise, and except expressing in endochylema, also has a large amount of nuclear expressions, and prompting NF-κ Bp65/I κ B α participates in generation, the development of PLGC.After application benefiting QI for activating blood circulation method and WEIFUCHUN are intervened, in high dose group, WEIFUCHUN group gastric mucosa, NF-κ B expresses obviously and reduces, and I κ B alpha expression obviously raises, and NF-κ B p65 absorbance and positive area rate are all starkly lower than model group.Point out benefiting QI for activating blood circulation ruling by law to express by lowering NF-κ B p65, rise I κ B α suppresses inflammatory cytokine (as IL-2, TNF-α) and transcribes, discharges, thereby alleviates mucosal inflammation reaction.Benefiting QI for activating blood circulation method has the effects such as blood circulation promoting and blood stasis dispelling, can improve gastric mucosa microcirculation, anti-oxidation stress, reduce the stimulations of cytokine to gastric mucosa such as ROS and TNF-α, thereby suppress the activation of NF-κ B, and make it cannot with the specific bindings such as TNF-α, the function that its controlling gene is transcribed is suppressed relevant, therefore alleviates the inflammatory reaction of gastric mucosa.
(5) cyclinD1, p16 albumen are in expression and the impact of benefiting QI for activating blood circulation method on it of PLGC rat stomach tissue
The formation of gastric cancer from the formation of tumor from biology angle see and be mainly manifested in: apoptosis obstacle on the one hand, cell infinite multiplication on the other hand, and follow dysdifferentiation.Cell proliferation is accelerated, and the cell of undergoing mutation can not be removed by Apoptosis Mechanism again, causes tumor to form.And the basic vital movements such as cell proliferation, differentiation, apoptosis and Association with Cell Cycle are close, therefore to see in this sense, tumor is the abnormal disease of a class cell cycle, cell cycle is the co-channel of cell proliferation and differentiation.
Cell order in cell cycle completes its propagation through G1-S-G2-M.Wherein G1 phase test point is particularly important, cell is integrated and is transmitted complicated cell internal/external signal at this, determine whether continue to enter proliferating cycle, after this point, cell does not just rely on born of the same parents' outgrowth adjustment signal and complys with independent procedure and postponed the whole cycle.Therefore, G1 time limit point processed is the key point of controlling Growth of Cells, determining cell fate.This adjusting function is abnormal, not only makes cell exogenous conditioning signal bradykinesia and is uncontrolled proliferation, and also cannot block has the DNA of damage to enter the S phase to copy, and then generates specific tumor phenotypes.Cyclin D1 (cyclinD1) is the shaping regulatory factor that in cell cycle, the G1 phase enters the S phase, is also considered to oncogene.CyclinD1 gene mapping, can be by the activity of combination active cell cyclin dependent protein kinase 4/6 (CDK4/6) in 11q13, and Phospho-Rb, makes cell enter the S phase by G1/S monitoring point, completes copying of DNA, accelerates the propagation of cell.The overexpression of cyclinD1 can shorten the time of G1 phase or accelerate G1 phase process, and the detection that even can make cell flee from test point directly enters the S phase, causes cell growth out of control.CyclinD1 is the key protein of G1 phase cell proliferation signal.Research finds that cyclinD1 exists gene amplification and protein overexpression in gastric cancer, but irrelevant with Tumor Differentiation degree, and this prompting cyclinD1 abnormal expression may be early stage molecular events in gastric cancer generation evolution.
P16 antioncogene claims again CDK41 gene, that first is found to directly act on cell cycle, suppress fissional antioncogene, it is a kind of cyclin dependant inhibitive factor (CDKI), it plays negative regulation function by the active cell cycle that suppresses cell cycle protein dependent kinase (CDKs), be positioned 9p21, in regulation of cell proliferation, p16 gene is that the effect that suppresses cell cycle mainly realizes by cyclinS-CDK4,6-pRb-E2F approach.The combination of p16 albumen and cyclinD1 competition and CDK4/6, loses the protein kinase activity of cyclinD1/CDK4/6 complex, to regulate and control G1-S phase stuck point, thereby stop, control cell division, Cell differentiation inducing activity, performance negativity regulating action, completes the regulation and control on cell proliferation cycle jointly.But while causing the improper expression of p16 albumen when p16 protein delation, sudden change, cell proliferation, by out of control, causes tumor to occur.Be starkly lower than normal gastric mucosa and non-cancerous issue at 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 interacts, and is keeping dynamic equilibrium, by regulating the functional status of Rb albumen to affect the process of G1 phase.Much research shows that the activation of NF-κ B path can promote the growth of gastric cancer and other tumors, and NF-κ B can, by regulate it to express in conjunction with the promoter of cyclinDs, regulate the growth of gastric cancer.
This research finds that normal rats hepatocyte has a small amount of cyclinD1 and p16 positive expression, express with cytoplasm and nucleus, model group liver tissues of rats all has cyclinD1 and p16 high expressed, positive expression is in nucleus, all compared with normal group obviously raises, difference has very significant (P < 0.01, P < 0.05).Showing that cyclinD1 and p16 have participated in the evolution of PLGC, may be the early stage event that gastric cancer occurs.After application benefiting QI for activating blood circulation method and WEIFUCHUN are intervened, high dose group and WEIFUCHUN group gastric mucosa organize cyclinD1 to express significantly decline, and benefiting QI for activating blood circulation high dose group p16 raises significantly.Show that benefiting QI for activating blood circulation method may activate NF-κ B signal path inhibition cyclinD1 by reducing pro-inflammatory cytokine, raises p16 and expresses control gastric precancerous lesion.
But signal transduction pathway is a complicated network, between each passage, influence each other, association, exist phosphorylation and dephosphorylation balance, the balance regulation mechanism of short apoptosis and apoptosis inhibit, once these balances or regulatory mechanism are broken, just may cause pathological process.The mechanism of each bars Signal Transduction Pathways and these Pathway Activations will be emphasis and the focus of PLGC research.
The present invention improves degree of inflammation and the atrophy degree of gastric tissue, alleviates mucosa tissue hyper-proliferative; Regulate cytokine network, improve TNF-α activity, reduce IL-6, IL-8 generation; Suppressing key protein NF-κ B in gastric tissue NF-κ B/I κ B alpha signal pathway expresses and activation; Suppress the expression 1 of gastric tissue cyclinD, raise p16 and express control gastric precancerous lesion.
Claims (3)
1. a compound stomach medicine for benefiting QI for activating blood circulation, is characterized in that, comprises Radix Codonopsis 20g, Radix Astragali 30g, Fructus Amomi 9g, Radix Notoginseng 9g, Radix Angelicae Sinensis 20g, Fructus Citri Sarcodactylis 10g, Rhizoma Chuanxiong 15g, Rhizoma Curcumae 12g, counts 125 grams/subsides, totally 200 pastes.
2. the compound stomach medicine of a kind of benefiting QI for activating blood circulation according to claim 1, is characterized in that, its preparation method, by above-mentioned medical material, is soaked 0.5 hour, decocts 2 times, adds for the first time 10 times of water gagings, boils and extracts 1.5 hours; Add for the second time 8 times of water gagings, boil and extract 1.0 hours.After merging extracted twice liquid, be concentrated into 6000 milliliters, fill to 500 mL of saline bottle, scarfing, sterilization, cold preservation is for subsequent use.
3. compound stomach medicine of a kind of benefiting QI for activating blood circulation according to claim 1 and preparation method thereof, middle dosage: 21g/kg/d, by the low middle Senior Three dosage group of ratio setting of 1: 2: 4, high dose group is: 42g/kg/d, general fluid extract is all prepared into high dose group, when administration, appropriateness dilution is used, press the dosage of 1mL/100g/d body weight, be 10mL/kg/d, the crude drug content of crude drug content=(42g/kg/d)/(10mL/kg/d)=4.2g/mL is high dose group fluid extract concentration, extractum amount=25000g/ (4.2g/mL)=5952mL, about 6000mL.
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| WO2020009949A1 (en) * | 2018-07-05 | 2020-01-09 | Boston Scientific Scimed, Inc. | Devices, systems, and methods for determining inflammation and/or fibrosis |
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| CN115487247A (en) * | 2022-11-04 | 2022-12-20 | 杭州市中医院 | Traditional Chinese medicine composition for treating threatened abortion and process |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105126012A (en) * | 2015-07-23 | 2015-12-09 | 申金玉 | Traditional Chinese medicinal preparation for treating gastric cancer and preparation method of traditional Chinese medicinal preparation |
| CN106620062A (en) * | 2016-11-09 | 2017-05-10 | 江苏金氏丹科技有限公司 | Traditional Chinese medicinal composition for protecting gastric mucosa |
| WO2020009949A1 (en) * | 2018-07-05 | 2020-01-09 | Boston Scientific Scimed, Inc. | Devices, systems, and methods for determining inflammation and/or fibrosis |
| US11058345B2 (en) | 2018-07-05 | 2021-07-13 | Boston Scientific Scimed, Inc. | Devices, systems, and methods for determining inflammation and/or fibrosis |
| CN113559232A (en) * | 2021-08-25 | 2021-10-29 | 朱玉辉 | Traditional Chinese medicine composition for enhancing gastrointestinal function and preparation method thereof |
| CN115487247A (en) * | 2022-11-04 | 2022-12-20 | 杭州市中医院 | Traditional Chinese medicine composition for treating threatened abortion and process |
| CN115487247B (en) * | 2022-11-04 | 2023-06-16 | 杭州市中医院 | Traditional Chinese medicine composition and process for treating threatened abortion |
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