CN106038765A - Application of compound Rabdosia rubescens preparation to chemical prevention of esophagus cancer - Google Patents

Abstract

The invention specifically relates to application of a compound Rabdosia rubescens preparation to chemical prevention of esophagus cancer, belonging to the technical field of tumor prevention. A formula for the compound Rabdosia rubescens preparation comprises 3 to 15 g of Rabdosia rubescens, 2 to 7.5 g of heydyotis diffusa and 2 to 7.5 g of barbat skullcap. The compound Rabdosia rubescens preparation is used for prevention of the esophagus cancer and esophageal precancerous lesion. According to the invention, an esophageal canceration rat model induced by NMBA is constructed at first, then concrete evaluation is carried out on the effect of a traditional Chinese medicine composition composed of the Rabdosia rubescens, heydyotis diffusa and barbat skullcap on chemical prevention of the esophagus cancer; and results show that although esophageal canceration is a multi-cause chronic continuous process, the traditional Chinese medicine composition composed of the Rabdosia rubescens, heydyotis diffusa and barbat skullcap, due to diversity of Chinese herbal medicine components, can exert prevention and treatment effect in the stage of development of the esophagus cancer, is capable of obviously inhibiting canceration of cells and presents good prevention effect.

Description

The application in esophageal carcinoma chemoprophylaxis of the Rabdosia rubescens compound preparation

Technical field

The invention belongs to tumor prevention technical field, be specifically related to a kind of Rabdosia rubescens compound preparation in esophageal carcinoma chemoprophylaxis In the patent application of application.

Background technology

The esophageal carcinoma is a kind of malignant tumor originating from epithelium of esophagus tissue, and it occurs development to be multifactor, polygenes, many Stage coefficient result, with age, sex, smoking, Diet lifestyle, living environment and inherited genetic factors phase all closely Close.Although the treatment meanss such as operation, chemotherapy and radiation have obtained relatively broad application in the treatment of the esophageal carcinoma, but esophagus Incidence rate and the mortality rate of cancer are not obviously improved.This suggests that we study esophageal carcinoma generation development mechanism right further It is one of Critical policies reducing epithelium of esophagus carcinogenesis rate that this process carries out intervening.

It is by atypical hyperplasia normal, slight, moderate atypical hyperplasia, severe that people's squamous cell carcinoma of esophagus becomes process Atypical hyperplasia, cancer in situ finally arrive the dynamic pathological change process of infiltrating carcinoma.Before pathological changes before cancer is referred to as cancer in position Pathological changes, takes around 5-10 from the process that normal cell turnover is cancerous cell, and during this, epithelial cell can continue to develop into Cancerous cell, it is also possible to stop differentiation and be in steady statue, it might even be possible to reversing and become normal cell, it has unstable and two-way The characteristic converted, this just intervenes the esophageal carcinoma for us and occurs the process of development to provide theoretical basis.

The chemoprophylaxis of cancer refers to that utilizing naturally occurring or synthetic chemical substance to stop, slow down or reverse cancer occurs Evolution, thus reduce cancer incidence and mortality rate.Chinese herbal medicine is as Chinese distinctive medicine resource, and it is at oncotherapy In effect more and more paid attention to, and the most more existing applied research.But with regard to Chinese medicine medicine in disease prevention For reason, it apparently more agrees with the prophylactic applications of tumor, thus pole is necessary Chinese herbal medicine or its compound formulation that exploitation is relevant Application in the chemoprophylaxis of cancer.

For the Rabdosia rubescens in Chinese traditional herbs, Herba Scutellariae Barbatae, Herba Hedyotidis Diffusae etc., its answering in kinds of tumors is treated Use existing preliminary study, such as: recording according to " China's oncotherapy great achievement ", Herba Scutellariae Barbatae cures mainly primary hepatocarcinoma, gastric cancer, rectal cancer etc. The gynecological tumors such as digestive tract tumor and nasopharyngeal carcinoma, pulmonary carcinoma and cervical cancer；Herba Hedyotidis Diffusae can be used for treat gastric cancer, rectal cancer, Alimentary tract cancer and the lymphsystem tumors such as malignant lymphoma, pulmonary carcinoma, cervical cancer, ovarian cancer, bladder cancer；And Rabdosia rubescens is disappearing Change in the research that road tumor especially treats the esophageal carcinoma, obtained definite curative effect and be widely recognized as；But exist about these Chinese herbal medicine Effect in the chemoprophylaxis of cancer still lacks more systematic research.

Summary of the invention

Present invention is primarily intended to the application that have studied Rabdosia rubescens compound preparation in esophageal carcinoma chemoprophylaxis, for the esophageal carcinoma And the chemoprophylaxis of precancerous lesion provides new possibility, the most also provide for the application in cancer prevention of other Chinese herbal medicine Reference and reference.

The detailed technology scheme that the present invention is taked is as described below.

The application in esophageal carcinoma chemoprophylaxis of the Rabdosia rubescens compound preparation, for the prevention of alimentary tract cancer, can suppress and Delay the Carcinogenesis of esophagus, the precancerous lesion process of normal epithelium of esophagus is played elemental abundances；Further concrete and Speech, the epithelium of esophagus precancerous lesion induced by NMBA for prevention；

Described Rabdosia rubescens compound preparation proportioning is: Rabdosia rubescens 3 ~ 15g, Herba Hedyotidis Diffusae 2 ~ 7.5g, Herba Scutellariae Barbatae 2 ~ 7.5g；Preferably join Ratio is: Rabdosia rubescens 10g, Herba Hedyotidis Diffusae 5g, Herba Scutellariae Barbatae 5g.

Time specifically used, as a example by a 60kg adult, every secondary preparation (Rabdosia rubescens 10g, Herba Hedyotidis Diffusae 5g, Herba Scutellariae Barbatae 5g) add decocting and become 400mL, sooner or later respectively take 200mL, can preferably prevent the formation of digestive tract tumor, there is suppression simultaneously and prolong The effect of slow canceration course of esophageal.

Rabdosia rubescens (Rabdosia rubescens(Hemsl1) Hara) it is Labiatae Rabdosia plant, containing multipleization Study point, the effect such as there is clearing away heat and expelling pathogen in the exterior, anti-inflammatory analgetic, stomach invigorating are invigorated blood circulation.Study and thought, the anticancer component master in Rabdosia rubescens If rubescensine A and rubescensine B, ehrlich ascites carcinoma, hepatocarcinoma, lymphoma, cerebroma, reticulum cell sarcoma etc. are respectively provided with substantially Antitumor action.

Herba Scutellariae Barbatae (Scutellaria barbata D.Don), belong to Labiatae HUANGDA platymiscium, call narrow leaf Herba Scutellariae indicae, And head grass, toothbrush grass etc., with dry all herbal medicine；Acrid in the mouth, hardship, cold in nature, return lung, liver, kidney channel.Complex chemical composition, mainly contains There are flavonoid and diterpene-kind compound.There is heat-clearing and toxic substances removing, dissipate addiction hemostasis, effect of inducing diuresis to remove edema, be used for treating pyogenic infections from tumour or sore, throat Swell and ache, venom, tumbling and swelling and edema, yellow cellulitis.

Herba Hedyotidis Diffusae (Hedyotis (Oldenlandia) diffusa (Willd.) Roxb.), annual hang down loosely Draft, genus rubia section plants of Hedyotis, call Herba Hedyotidis Diffusae, Serpentis Herba Urticae Cannabinae, Herba Turczaninowiae Fastigiatae etc..Main component have flavone and flavonol glycosides, Terpenoid.Bitter sweet, cold, primary efficacy has heat-clearing and toxic substances removing, promoting blood circulation and detumescence, and pain relieving is antibacterial, among the people for treating intestinal tract disease, poisonous snake The diseases such as medicine wound.Clinically, Herba Hedyotidis Diffusae can be used for the treatment of kinds cancer, such as: gastric cancer, rectal cancer, malignant lymphoma, Pulmonary carcinoma, cervical cancer, ovarian cancer, bladder cancer, primary hepatocarcinoma, nasopharyngeal carcinoma etc. and lymphsystem tumor.

For Rabdosia rubescens, Herba Scutellariae Barbatae and Herba Hedyotidis Diffusae these three Chinese herbal medicine, prior art be mostly for its individually or With other drug combination applied research in related neoplasms, application is then lacked for the combination application of these three Chinese herbal medicine and grinds Study carefully.It addition, in prior art for Chinese herbal medicine related neoplasms treat in applied research, typically mostly be based on cell aspect Preliminary study, and lack certain the depth of investigation.

The application being configured to related drugs of animal tumor model and the research of the mechanism of action provide preferable basis, pin For the esophageal carcinoma, Methyl jasmonate (N-nitrosomethylbenzylamine, NMBA) rat esophagus induced Canceration model, owing to can fully simulate people's carcinogenesis of esophageal epithelium, thus can be that esophageal canceration Mechanism Study is effective with screening Intervene medicine and good platform is provided.The present invention is i.e. on this Research Thinking, first passes through and builds the rat induced by NMBA Esophageal canceration model, then to the compound Chinese medicinal preparation being made up of Rabdosia rubescens, Herba Scutellariae Barbatae and Herba Hedyotidis Diffusae at esophagus cancerization The effect learned in prevention has carried out concrete evaluation.Result shows, although the canceration of the esophageal carcinoma is a multi-pathogenesis, chronic, company The process of continuous property, but due to the multiformity of medicinal herb components, Rabdosia rubescens, Herba Scutellariae Barbatae and Herba Hedyotidis Diffusae in the present invention are constituted Compound Chinese medicinal preparation can play prevention and the effect for the treatment of in each stage that the esophageal carcinoma occurs, it is possible to substantially suppression canceration is thin Intracellular growth, shows preferable preventive effect.Plus the low raw-material cost of Chinese herbal medicine, and toxicity is little, non-evident effect Advantage so that the compound Chinese medicinal preparation being made up of Rabdosia rubescens, Herba Scutellariae Barbatae and Herba Hedyotidis Diffusae of the present invention food Having preferable application prospect in the chemoprophylaxis of pipe cancer, the most also the research and development for other similar compound Chinese medicinal preparation provide relatively Good reference and reference function.

Accompanying drawing explanation

Fig. 1 is experiment packet situation schematic diagram；

Fig. 2 is rat esophagus mucosa HE dyeing pathological grading figure (× 400), and wherein A is NMBA group rat esophagus stage casing pathology figure, B is typical cytopathic enlarged drawing, and wherein a is normal esophageal mucosa, and b is mucous membrane of esophagus proliferative lesion, and c is mucous membrane of esophagus white macula Pathological changes, d is mucous membrane of esophagus atypical hyperplasia；

Fig. 3 is rat dietary intake, drinking-water, weight data curve, every cage (5) rat during wherein A is 35 weeks during experiment Average dietary intake amount weekly；B is that during 35 weeks, every cage (5) rat the most averagely takes in amount of drinking water；C is during 35 weeks every Rat (5, every cage) body weight change amount weekly；

Fig. 4 is the esophagus HE stained statistical result of each time point (15w, 25w, 35w) rat；

Fig. 5 is the statistical result of the inhibition of the rat esophagus carcinogenesis that NMBA is induced by Rabdosia rubescens compound preparation group；

Rat esophagus mucosa squamous epithelial cancer Ki67 expression statistical result is respectively organized when Fig. 6 is 35w；Wherein a is solvent control group, B is Rabdosia rubescens compound preparation matched group, and c is Methyl jasmonate group, and d is zengshengping tablet group, and e is low dose of Rabdosia rubescens compound preparation Amount group, f is Rabdosia rubescens compound preparation high dose group；

Rat esophagus mucosa squamous epithelial cancer GSK-3 β expression statistical result is respectively organized when Fig. 7 is 35w；Wherein a is solvent control Group, b is Rabdosia rubescens compound preparation matched group, and c is Methyl jasmonate group, and d is zengshengping tablet group, and e is Rabdosia rubescens compound preparation Low dose group, f is Rabdosia rubescens compound preparation high dose group；

Rat esophagus mucosa squamous epithelial cancer β-catenin expression statistical result is respectively organized when Fig. 8 is 35w；Wherein a is solvent pair According to group, b is Rabdosia rubescens compound preparation matched group, and c is Methyl jasmonate group, and d is zengshengping tablet group, and e is Rabdosia rubescens compound recipe system Agent low dose group, f is Rabdosia rubescens compound preparation high dose group；

Rat esophagus mucosa squamous epithelial cancer c-jun expression statistical result is respectively organized when Fig. 9 is 35w；Wherein a is solvent control Group, b is Rabdosia rubescens compound preparation matched group, and c is Methyl jasmonate group, and d is zengshengping tablet group, and e is Rabdosia rubescens compound preparation Low dose group, f is Rabdosia rubescens compound preparation high dose group.

Detailed description of the invention

Below in conjunction with embodiment, the application is further explained explanation, before introducing specific embodiment, in the present invention Partial material situation is briefly discussed below.

Raw material of Chinese medicine: in following embodiment, Rabdosia rubescens used is the dry stem and leaf (aerial parts) of Rabdosia rubescens, and Herba Scutellariae Barbatae is It is dried herb；Herba Hedyotidis Diffusae, for being dried herb, is purchased from No.1 Hospital Attached to He'nan Traditional Chinese Medical College；

Experimentation Chinese medicine consumption standard is, with reference to adult's dosage standard in " Chinese Pharmacopoeia " (2015 editions), by its 7 times Dosage calculate the dosage (reference body surface area conversion mode, the dosage of people is 7 times of rat dosage) for mice；

In following embodiment, Traditional Chinese medicine decocting method is (as a example by a secondary dosage, it may be assumed that Rabdosia rubescens 70g, Herba Hedyotidis Diffusae 35g, Herba Scutellariae Barbatae 35g) be:

Medicine to be processed is loaded in special non-woven bag, mouth is tied, puts in container, cold water soak 30 minutes；

Employing boiling machine decocts, and decocts about about 20min, and during decoction, the ratio of adding water is, and the amount of liquid medicine of amount of water=required+ 20%；

It is fried that associative operation is functioned in an acting capacity of operation according to a conventional method by Chinese medicine pharmacy of No.1 Hospital Attached to He'nan Traditional Chinese Medical College.

Laboratory animal: rat F344(Fischer Rats, 7 ~ 8 week old), SPF level, body weight is 90g to 120g, and the quality certification is numbered: 11400700078563, it is purchased from Beijing Vital River Experimental Animals Technology Co., Ltd.；

In experimentation, rat is raised in barrier system less than 5 by every cage；Feeding environment: temperature, 20 ± 2 DEG C；Relatively Humidity, 50 ± 10%；Periodicity of illumination, 12h day/night, through autoclaved SPF level animal feed, bedding and padding and single steaming water, The free diet of rat.

Embodiment

Before being further described embodiment, it is to be understood that for chemoprophylactic related experiment, concrete pre-preventive effect Fruit is accompanied by (Induction Process in other words) of corresponding Carcinogenesis, thus the effect of Chinese herbal compounds of the present invention Fruit is also accompanied by the rat esophagus Carcinogenesis of NMBA induction and carries out, and related experiment process is briefly discussed below.

One, experiment packet and experimentation

In order to verify used Chinese medicine composition elemental abundances in the rat esophagus Carcinogenesis that NMBA induces, first First need rat is suitably grouped (rat buy after first adaptability feed 2 weeks be grouped again), packet situation is as described below:

After rat adaptability feeds 2 weeks, by laboratory animal random packet, packet situation as it is shown in figure 1, particularly as follows:

Solvent control group (n=20), conventional raising, compare with normal saline gavage (gavage volume 10 ml/kg) conduct every day, When NMBA induces 15w, 25w, 35w, take 5,5,10 rats respectively, dissect rat after anesthesia and take food pipe observation and locate further Reason；

Rabdosia rubescens compound preparation matched group (n=5), conventional raises, every day high dose Rabdosia rubescens compound preparation gavage (gavage Volume 10mL/kg) as Chinese medicine matched group, start gavage, continuous gavage 35w from-1w, dissect whole 5 rats after anesthesia, take Esophagus is observed and processes further；

NMBA model group (n=33), subcutaneous injection NMBA(0.5mg/kg), 3 times a week, it is administered 5w, conventional raising altogether；At NMBA During induction 15w, 25w, 35w, take 10,10,13 rat anesthesias respectively and dissect, take food pipe and observe and process further；

Zengshengping tablet is as positive controls (n=10), subcutaneous injection NMBA(0.5mg/kg), 3 times a week, it is administered 5w altogether, simultaneously Every day with zengshengping tablet to rat oral gavage (being equal to the dose,equivalent 4mg/kg of people, gavage volume 10mL/kg), every day 1 time, conventional Raise；When NMBA induces 35w, after anesthesia, dissect whole 10 rats, take food pipe and observe and process further；Zengshengping tablet (specification: 0.33g × 80s, manufacturer: Tianjin Central Pharmaceutical Co., Ltd), as a kind of compound Chinese medicinal preparation, it is main Raw material has Radix Sophorae Tonkinensis, Rhizoma Bistortae, Sonchus brachyotus DC., Spica Prunellae, Cortex Dictamni, Rhizoma Dioscoreae Bulbiferae；There is effect of heat-clearing and toxic substances removing, blood stasis-eliminating and stagnation-dissipating；Its The scope of application includes: is applicable to esophagus and cardia epithelial proliferation, has singultus, dysphagia of taking food, xerostomia, bitter taste, pharyngalgia, just Dry tongue is dark, the hot stasis of blood of people with thready and slippery pulse interior knot shower；Anterogastric epithelial hyperplasia that carcinogen is induced, hyperplasia of oral mucosa, nose Pharynx epithelial proliferation and mucous membrane of urinary bladder hypertrophy are inhibited, can improve the immunologic function of body；The most extensively should With, it is the medicine of the suppression esophageal epithelial proliferation the most relatively generally acknowledged, thus in this, as positive control；

Rabdosia rubescens compound preparation high dose group (n=15), subcutaneous injection NMBA(0.5mg/kg), 3 times a week, start to fill from-1w Stomach, is administered 5w altogether, (is equal to the dose,equivalent of people, gavage volume with Rabdosia rubescens compound preparation to rat oral gavage every day simultaneously 10ml/kg), every day 1 time, conventional raising；NMBA induce 35w time, dissect whole 15 rats after anesthesia, take food pipe observe with And process further；

Rabdosia rubescens compound preparation low dose group (n=15), subcutaneous injection NMBA(0.5mg/kg), 3 times a week, start to fill from-1w Stomach, is administered 5w altogether, simultaneously every day with Rabdosia rubescens compound preparation to rat oral gavage (the Rabdosia rubescens high dose of 40%, gavage volume 10mL/kg), every day 1 time, conventional raising；NMBA induce 35w time, dissect whole 15 rats after anesthesia, take food pipe observe with And process further.

Each process group records weekly a rat amount of drinking water and dietary intake amount during being administered, it is big that every day measures each experimental group Mus body weight.

It should be noted that rat esophagus observes sampling and evaluation methodology is as described below in experimentation:

(1) esophagus sampling

Rat injecting narcotic, to after the whole deliquescing of health, is lain on the back rat and is fixed on operating-table, and general abdominal is with 75% ethanol Embrocate sterilization, upwards cut off thoracic cavity to bottleneck throat with shears along rat abdomen median line, find stomach in abdominal cavity, start edge from stomach cardia Esophagus upwards with connective tissues such as anodontia tweezers and little shears careful separation esophagus peripheral adipose, takes rat stomach pars cardiaca to throat The esophagus in portion；

Taken food pipe is positioned on blank sheet of paper, cuts off along the esophagus longitudinal axis with shears, at the beginning of rat esophagus mucosa infection situation is carried out The morphological observation of step；

Then being divided into two by esophagus along the longitudinal axis and be uniformly divided into two parts, be more each divided into upper, middle and lower segment, a part is put Entering equipped with fixing (24h) in the 2mL centrifuge tube of 4% paraformaldehyde, routine paraffin wax embeds, and makes the section that thickness is 4 μm, and HE contaminates Color；Second half is directly placed into-80 DEG C of cryogenic refrigerators and preserves, it is simple to checks or detects sundry item.

(2) optical microphotograph Microscopic observation record rat esophagus epithelial pathological tissue classification

By two pathologists under double blinding state, to the HE stained of rat esophagus in light Microscopic observation esophagus pathologic group Knitting situation of change and record the number of different lesions type under each visual field, the pathological grading standard of epithelium of esophagus is with reference to 1979 Professor and the Gary D. professor Stoner method to rat esophagus lesion type in Nian Linpei, HE dyeing pathological grading standard is such as Shown in Fig. 2, particularly as follows:

Normal: normal esophageal mucosa is without thickening, and squamous cell marshalling, without increasing；

Proliferative lesion: Esophageal Mucosa thickens, basal layer cell increases, epithelial cell (spine cell) increases, and thickens place mucosa row Arrange the most random；

Leukoplakia: mucous membrane of esophagus cuticular layer thickens substantially, and basal cell and spine cell significantly increase, the basal cell of hypertrophy Form more than 3-6 layer, the 1/2 of holostrome can be accounted for, arrangement disorder；

Atypical hyperplasia: Esophageal Mucosa basal layer cell hypertrophy is obvious, and level is disorderly and more, basad portion highlights, and can exceed The 2/3 of epithelium holostrome.Cell increases, and size and form are inconsistent, and mucous layer cuticular layer all thickens.

(3) preventive effect evaluation result Rabdosia rubescens compound preparation for esophageal canceration chemoprevention efficacy evaluate mainly from Histopathology aspect is carried out, and relevant evaluation process and result are briefly discussed below.

1, upgrowth situation evaluation in experimentation

Preliminary observation and statistical result show, in whole experimentation, each group rat mental status is good, rat amount of drinking water and Dietary intake amount difference is little, no significant difference (P ＞ 0.05)；Rat body weight grows steadily, and respectively tests group difference Not statistically significant (P ＞ 0.05) (the most as shown in Figure 3).

2, pathologic examination situation

(1) the naked eyes preliminary morphological observation to rat esophagus mucosa infection situation in anatomic course

At 3 time points (NMBA lures cancer to start rear 15w, 25w, 35w) of whole experiment, compared with matched group, NMBA induced module The esophagus of type group rat is without substantially increasing thick phenomenon, and mucosa is the most glossy, and seriality is preferable, has no obvious pathological changes.

(2) in the case of observation by light microscope, NMBA guidance model group rat esophagus mucosa and the group of squamous cell Knit feature

It is said that in general, rat esophagus carcinogenesis process is by normal, hypertrophy, leukoplakia, atypical hyperplasia, papilloma After be converted into cancer in situ, infiltrating carcinoma.In this experiment, for building rat esophagus canceration model, have employed subcutaneous injection NMBA induction The generation of canceration course of esophageal, the histological change situation for determining inducing effect, to the rat esophagus of NMBA guidance model group It is briefly discussed below.

Observation indicate that: at each time point (luring cancer to start rear 15w, 25w, 35w) of whole experiment, matched group esophagus Mucous layer surface is Normal squamous epithelium, and its basal layer cell is monolayer alignment, without thickening and increasing performance；And NMBA induced module Mucosa squamous cell edema seen from type group, basal layer cell increases and thickens, and the proper mucous membrane of squamous epithelial cancer upwards prolongs Under supreme epidermis layer, lamina propria edema thickens, the nucleus increase of visible hyperplasia under high power lens (× 400), form, not of uniform size Etc. phenomenon, mucous membrane of esophagus occurs that the cancerations such as hypertrophy, leukoplakia and atypical hyperplasia in various degree occur, but mucous membrane of esophagus Substrate and muscular tissue are showed no cancer in situ and infiltrating carcinoma.

Esophagus HE stained (mucous membrane of esophagus pathological changes) statistical result such as Fig. 4 of each time point (15w, 25w, 35w) rat Shown in；Compared with matched group, NMBA group is hypertrophy, leukoplakia and atypical hyperplasia pathological changes showed increased when 15w, 25w, 35w, Difference statistically significant (P < 0.05)；Hypertrophic change: the obvious hypertrophy of 35w mucous membrane of esophagus, pathological changes increases, with 15w, 25w phase Ratio is variant, difference statistically significant (P < 0.001)；Compared with atypical hyperplasia: 35w with 15w, 25w, lesion degree is serious, poor Different statistically significant (P < 0.05).

The number of cells of various pathological changes under visual field each during optical microphotograph Microscopic observation is recorded and adds up, In the F344 rat esophagus carcinogenesis evolution process of NMBA induction, each pathological changes (hypertrophy, leukoplakia, atypical hyperplasia) is individual Number (mean ± standard deviation) result is as shown in the table:

Note: a represents compared with matched group, and when 15w, 25w, 35w, NMBA group hypertrophy, leukoplakia and atypical hyperplasia pathological changes are obvious Increase, difference statistically significant (P< 0.05)；Proliferative lesion: when 25w, 35w, hypertrophy substantially increases compared with 15w NMBA group Many, difference statistically significant (P< 0.05)；Atypical hyperplasia showed increased, difference when 25w, 35w compared with 15w NMBA group Statistically significant, (P< 0.05).

The observed result of summary optical microscope and statistical result, it can be clearly seen that, NMBA induce 15w, When 25w, 35w, in epithelium of esophagus, the pathological changes showed increased such as hypertrophy, leukoplakia and atypical hyperplasia, the result shows NMBA success is induction of rat esophagus carcinogenesis process, it is believed that epithelium of esophagus canceration guidance model successfully constructs.

3, in the case of observation by light microscope, Rabdosia rubescens compound preparation group rat esophagus mucosa and the group of squamous cell Knit feature

Preliminary morphological observation to the rat esophagus epithelium of corresponding time point (35w), result shows, Rabdosia rubescens compound preparation group Similar with the mucous membrane of esophagus of positive controls rat, no significant difference between pathological changes.

The inhibition (mucous membrane of esophagus pathological changes) of the rat esophagus carcinogenesis that NMBA is induced by Rabdosia rubescens compound preparation group Statistical result as shown in Figure 5.From fig. 5, it can be seen that each experimental group pathological changes (hypertrophy, leukoplakia, atypical hyperplasia) all with NMBA group is compared, and its result is:

Hypertrophy, Rabdosia rubescens low dose group difference statistically significant (P < 0.05), zengshengping tablet group and Rabdosia rubescens high dose difference without Meaning；

Leukoplakia, zengshengping tablet group, Rabdosia rubescens low dose group and Rabdosia rubescens high dose group difference the most statistically significant (P < 0.000)；

Atypical hyperplasia, zengshengping tablet group and Rabdosia rubescens high dose difference are statistically significant, and Rabdosia rubescens low dosage difference is without statistics Learn meaning (* represents that P < 0.05, * * represents that P < 0.01, * * * represents P < 0.001).

4, the evaluation of overall preventive effect

After experiment terminates, to the number of each group every rat esophagus pathological changes (hypertrophy, leukoplakia, atypical hyperplasia) (mean ± Standard deviation) to add up, result is as shown in the table:

Note: a represents compared with NMBA group, and difference statistically significant (P < 0.05), b represents that difference has system compared with NMBA group Meaning (P < 0.01) learned by meter, and c represents compared with NMBA group, difference statistically significant (P < 0.001).

As can be seen from the above table, zengshengping tablet group, Rabdosia rubescens compound preparation low dose group, Rabdosia rubescens compound preparation high dose group Pathological changes number all reduces (P < 0.05) compared with NMBA group.It is big that this result explanation compound ' Donglingcao ' preparation inhibits that NMBA induces Mus mucous membrane of esophagus hypertrophy, leukoplakia and atypical hyperplasia pathological changes.

Further, to respectively organizing rat esophagus mucosa squamous epithelial cancer Ki67, GSK-3 β, β-catenin, c-jun during 35w Expression is added up, and result is as shown in Fig. 6 ~ 9.It can be seen that compared with NMBA group, compound ' Donglingcao ' preparation group (including that high dose combines low dose group) is respectively provided with preferable technique effect, suitable with zengshengping tablet group, and even part is better than hypertrophy Flat group, show certain application prospect.

Claims (3)

1. Rabdosia rubescens compound preparation application in esophageal carcinoma chemoprophylaxis, it is characterised in that before the esophageal carcinoma and the esophageal carcinoma The prevention of pathological changes,

Described Rabdosia rubescens compound preparation proportioning is: Rabdosia rubescens 3 ~ 15g, Herba Hedyotidis Diffusae 2 ~ 7.5g, Herba Scutellariae Barbatae 2 ~ 7.5g.

2. Rabdosia rubescens compound preparation application in esophageal carcinoma chemoprophylaxis as claimed in claim 1, it is characterised in that Rabdosia rubescens Compound preparation proportioning is: Rabdosia rubescens 10g, Herba Hedyotidis Diffusae 5g, Herba Scutellariae Barbatae 5g.

3. Rabdosia rubescens compound preparation application in esophageal carcinoma chemoprophylaxis as claimed in claim 1, it is characterised in that for pre- The anti-epithelium of esophagus cancer induced by NMBA.