CN109106893B

CN109106893B - Traditional Chinese medicine compound preparation for regulating intestinal barrier function and preparation method thereof

Info

Publication number: CN109106893B
Application number: CN201811340664.7A
Authority: CN
Inventors: 钟薏; 付淑娟; 吴婷婷; 周张杰; 杨蕴; 张士强
Original assignee: SHANGHAI TCM-INTEGRATED HOSPITAL
Current assignee: SHANGHAI TCM-INTEGRATED HOSPITAL
Priority date: 2018-11-12
Filing date: 2018-11-12
Publication date: 2021-04-20
Anticipated expiration: 2038-11-12
Also published as: CN109106893A

Abstract

The invention relates to a traditional Chinese medicine compound preparation for improving intestinal barrier function of tumor patients and regulating organism immunity and a preparation method thereof. The traditional Chinese medicine compound preparation is prepared from the following traditional Chinese medicine raw materials in parts by weight: 30-45 parts of astragalus membranaceus, 10-30 parts of codonopsis pilosula, 10-20 parts of poria cocos, 10-20 parts of bighead atractylodes rhizome, 10-15 parts of rhizoma smilacis glabrae, 10-30 parts of chinaroot greenbrier, 10-20 parts of fiveleaf akebia fruit, 5-20 parts of endothelium corneum gigeriae galli and 10-30 parts of Chinese actinidia root. The traditional Chinese medicine compound preparation has the effects of tonifying qi and spleen, eliminating dampness and resolving masses, can protect intestinal barrier injury and improve the life quality of patients in tumor patients with spleen deficiency as a main symptom caused by chemotherapy, has the advantages of low cost, safety, effectiveness, no toxic or side effect, and can be used for regulating PI3K/AKT expression in intestinal tumor microenvironment and remarkably reducing colorectal cancer postoperative metastasis.

Description

Traditional Chinese medicine compound preparation for regulating intestinal barrier function and preparation method thereof

Technical Field

The invention relates to the field of traditional Chinese medicines, in particular to a traditional Chinese medicine compound preparation for improving intestinal barrier function of tumor patients and regulating organism immunity and a preparation method thereof.

Background

Colorectal cancer is the fifth malignant tumor of the overall incidence rate in China, is the 3 rd among the gastrointestinal malignant tumors, and has a rising trend in recent years. The latest CONCORD-3 data shows that the five-year survival rate of colon cancer in China is obviously lower than that in European and American countries such as the United states, Canada, Iceland, Norway and is at a middle-to-low level in Asian countries and regions. At present, the first choice of a radical treatment method for colon cancer is still surgical treatment, but patients suffer from factors such as venous return, anatomical parts and the like after surgery, and relapse and metastasis occur in different proportions. The 5-year survival rate of the non-metastatic colorectal cancer patients can exceed 75 percent, and the 5-year survival rate of the metastatic colorectal cancer patients is 10 percent. Recurrence and metastasis after radical treatment become the leading cause of death of colorectal cancer patients, and the long-term survival rate of the patients is seriously influenced.

Intestinal tumor itself and after operation are easy to cause intestinal mucosa barrier dysfunction, inflammation, intestinal flora imbalance and the like, which can cause the loss of immune monitoring function, the reduction of the immune function of patients, the occurrence and even aggravation of intestinal mucosa inflammation, and the reduction of intestinal permeability, thereby causing the metastasis or recurrence of malignant tumor. Chemotherapy is a first-line adjuvant therapy method after radical tumor surgery, and in order to kill tumor cells, chemical drugs are used to inhibit the proliferation, infiltration and metastasis of the tumor cells, so that residual tumor cells or lesions with a malignant tendency can be eliminated while primary lesions are treated, and the effect of preventing relapse and metastasis is achieved to a certain extent. However, chemotherapy drugs inevitably have considerable toxic and side effects on the body, wherein intestinal barrier damage is the toxic response of some anticancer drugs. Therefore, chemotherapy induced changes in the intestinal microenvironment, including intestinal flora disturbance and impaired intestinal barrier function, may further cause immune response changes such as immune evasion, and even systemic inflammatory responses. Therefore, the composition can protect the intestinal barrier function, prevent and treat intestinal injury caused by chemotherapy, improve symptoms of patients, regulate systemic immunity, and has certain influence on the curative effect of chemotherapy and the progress of malignant tumors.

The traditional Chinese medicine is different from the holistic concept and the concept of treatment based on syndrome differentiation of western medicine, and has good curative effect on treating postoperative recurrence and metastasis of colorectal cancer patients by making up the limitation and deficiency of western medicine therapy. The early clinical subjects prove that the traditional Chinese medicine compound preparation for improving the intestinal barrier function of the tumor patients and regulating the organism immunity has good curative effect on improving the intestinal dysfunction of the malignant tumor patients after chemotherapy, reducing the relapse and metastasis after the operation and improving the organism immunity. The traditional Chinese medicine composition is proved to be effective for patients with middle and late malignant tumor spleen deficiency chemotherapy in early clinical subjects of a subject group, can protect intestinal barrier function of patients with malignant tumor chemotherapy, improves clinical symptoms of the patients, reduces chemotherapy toxic and side effects, improves the life quality of the patients, and is good in safety. The basic experiments also show that the traditional Chinese medicine compound preparation for improving the intestinal barrier function of the tumor patients and regulating the immunity of the organism can protect the intestinal barrier and regulate the PI3K/AKT expression in the intestinal tumor microenvironment to inhibit metastasis by combining chemotherapy.

Disclosure of Invention

The invention aims to provide a Chinese medicinal compound preparation which is safe and effective, simple and convenient to prepare, and moderate in price and can improve the intestinal barrier function of a tumor patient and adjust the immunity of an organism.

The invention also aims to provide a preparation method of the traditional Chinese medicine compound preparation for improving the intestinal barrier function of the tumor patients and regulating the immunity of the organism.

The invention provides a traditional Chinese medicine compound preparation for improving intestinal barrier function and regulating organism immunity of tumor patients, which is prepared from the following traditional Chinese medicine raw materials in parts by weight: 30-45 parts of astragalus membranaceus, 10-30 parts of codonopsis pilosula, 10-20 parts of poria cocos, 10-20 parts of bighead atractylodes rhizome, 10-15 parts of rhizoma smilacis glabrae, 10-30 parts of chinaroot greenbrier, 10-20 parts of fiveleaf akebia fruit, 5-20 parts of endothelium corneum gigeriae galli and 10-30 parts of Chinese actinidia root.

The invention has rigorous, scientific and reasonable formula and accords with the theory of traditional Chinese medicine. The codonopsis pilosula is beneficial to tonifying spleen and lung, the poria cocos is beneficial to spleen and stomach, the bighead atractylodes rhizome is used for regulating the middle warmer, the three medicines are ministerial medicines and have the effects of tonifying qi and spleen, and the effect of tonifying qi of the monarch medicine astragalus is enhanced. On the basis of the four medicines, fiveleaf akebia fruit, glabrous greenbrier rhizome, chinaroot greenbrier rhizome and Chinese actinidia root are added as adjuvant medicines, the fiveleaf akebia fruit has the effects of harmonizing stomach, soothing liver, regulating qi, slightly activating blood circulation, softening hardness to dissipate stagnation, the glabrous greenbrier rhizome and the Chinese actinidia root are used for eliminating dampness and relieving toxicity of chemotherapy, the chinaroot greenbrier is used for promoting diuresis, removing turbidity, detoxifying and dissipating blood stasis, and the chicken's. The full prescription has the outstanding efficacy of tonifying qi and spleen, the qi tonifying in the prescription is assisted by evacuation, the qi tonifying is mainly dispersed, and the full prescription is used together with cold and warm herbs, but is not greasy. The formula can protect intestinal barrier injury and improve the life quality of patients in patients with spleen deficiency caused by chemotherapy, and has good clinical curative effect.

The raw material medicaments related by the invention are all natural Chinese herbal medicaments, are all Chinese medicaments collected in the first edition of the Chinese pharmacopoeia in 2000 years, and meet the quality requirements of the pharmacopoeia. Easily-accessible raw materials, simple preparation, wide medicinal sources, and low cost. The invention follows the medication principle of the prescription of the traditional Chinese medicine, and various component medicines are taken by decocting in water.

The invention also provides a preparation method of the traditional Chinese medicine compound preparation for improving the intestinal barrier function of a tumor patient and regulating the immunity of an organism, which comprises the following steps:

(1) weighing each traditional Chinese medicine raw material medicine according to the prescription amount for later use;

(2) taking half of the amount of each traditional Chinese medicine raw material medicine in the step (1), adding water, decocting twice, filtering, combining decoction, concentrating and drying to obtain dry extract, and crushing the dry extract to obtain extract fine powder;

(3) taking the other half amount of the raw materials of the traditional Chinese medicines in the step (1), crushing, sterilizing, mixing with the extract fine powder prepared in the step (2), further adding pharmaceutically acceptable auxiliary materials, and preparing the conventional preparation of the traditional Chinese medicine compound preparation by a conventional traditional Chinese medicine preparation method.

Wherein, when the first decoction in the step (2) is carried out: adding 3-8 times of water, soaking for half an hour, and decocting for 45 minutes; during the second decoction: adding 3-8 times of water, and decocting for 30 minutes.

Wherein, the sterilization method in the step (3) is Co60 irradiation sterilization, and the irradiation dose is 8 k.

The invention adopts various conventional preparation processes, adds forming auxiliary materials or flavoring auxiliary materials, and can prepare the traditional Chinese medicine compound preparation provided by the invention into pharmaceutical dosage forms such as granules, tablets, capsules, oral liquid and the like. The pharmaceutical excipients are selected from mannitol, lactose, sucrose powder, starch, magnesium stearate, aspartame, menthol and/or sweet orange oil essence.

In addition, the traditional Chinese medicine raw materials in parts by weight can also achieve the treatment effect of the invention by adopting a decoction prepared by a conventional decoction method.

Animal pharmacodynamic experiment results show that compared with simple chemotherapy, the length, width and crypt depth of villus on ileum epithelium after HE staining, and the serum DAO, DLA and IFABP of the traditional Chinese medicine compound preparation are obviously lower than those of a chemotherapy group. The results of an animal model inoculated in situ with the intestinal cancer of a nude mouse show that the combined chemotherapy of the formula can effectively control the size of a tumor body and effectively reduce the expression of PI3K and AKT protein, thereby reducing the formation of a microenvironment for tumor metastasis and reducing the chance of tumor recurrence and metastasis. Clinical research results such as Chinese medicine symptom integration, experimental indexes, quality of life index observation and the like show that: the traditional Chinese medicine compound preparation has obvious and reliable effect, mild medicine property and no toxic or side effect, can obviously reduce the D-LA serum content rise induced by chemotherapy, has obvious inhibiting effect on the increase of intestinal mucosa permeability caused by the chemotherapy, and protects the intestinal mechanical barrier function from being damaged; in addition, the formula can up-regulate serum contents of sIgA, IL-2 and IFN-gamma, play an immune protection role in intestinal tracts and improve the immune barrier function of the intestinal tracts. Can obviously reduce the colorectal cancer postoperative metastasis and improve the life quality of patients.

Description of the drawings:

FIG. 1 is a graph showing the comparison of the body weights of the nude mice of each group in the animal experiment, wherein the difference of the body weights of the nude mice of each group has no statistical significance

FIG. 2 and FIG. 3 show the tumor weight comparison of nude mice in each group (FIG. 2 shows the tumor weight comparison of each group, FIG. 3 shows the intestinal tumor of each group) in animal experiments, which shows that the combination of the two chemotherapeutics of the formula can obviously reduce the intestinal tumor tissue compared with the single use of 5-fluorouracil, and the curative effect of the combination is equivalent to that of other groups

FIG. 4 is a comparison graph of protein electrophoresis of western blot detection of intestinal tissue protein in animal experiments, which shows that the formulation can effectively reduce the expression of PI3K and AKT in combination with chemotherapy

FIGS. 5-8 are graphs showing the expression of AKT and PI3K mRNA in intestinal mucosa detected by Real-Time PCR in animal experiments (FIG. 5 is PI3K pathological HE (200 times), FIG. 6 is the result of detecting the expression of PI3K mRNA in intestinal tissues by PCR, FIG. 7 is the result of detecting the expression of AKT pathological HE (200 times), and FIG. 8 is the result of detecting the expression of AKT mRNA in intestinal tissues by PCR), which shows that the formula can effectively reduce the expression of PI3K and AKT by combining chemotherapy with the traditional Chinese medicine

FIG. 9 is an evaluation of ileal epithelial injury index of each group, and the ileal epithelial injury degree of the experimental group is obviously lower than that of other four groups, and the experimental group has the function of relieving intestinal injury caused by chemotherapeutic drugs

FIG. 10 is a graph of multiple sampling (10 spots) HE (. about.100-fold) analysis of lung and liver metastasis counts, showing that the method reduces tumor metastasis

Detailed Description

The beneficial effects of the medicine of the present invention are further illustrated by the following test examples, and the tests include animal pharmacodynamics experiments and clinical efficacy observation experiments of the compound preparation for improving intestinal barrier function and regulating body immunity of tumor patients.

Example 1 an experimental study of the Chinese herbal compound preparation of the present invention for protecting intestinal barrier and regulating PI3K/AKT expression inhibition transfer 1. animal: 65 BALB/c female nude mice 4-5 weeks old, 30 NOD-SCID mice, 30 BALB/c nude mice, and 20-24 g of body mass, which are purchased from Shanghai Si Laike laboratory animals Co., Ltd, of Chinese academy, and are bred under SPF-level environment, and the qualification number of the laboratory animals is as follows: SYXK (Shanghai) 2014-0008.

2. The method comprises the following steps:

2.1 cell culture and subcutaneous transplantation of tumors

And (3) culturing the WT cells by using a DMEM medium, digesting and passaging by using 0.25% trypsin and 0.02% EDTA solution when the cells grow to 70% -80% fusion, and diluting to the cell number of 1.25 multiplied by 107ml < -1 > after counting. After the skin of each nude mouse is disinfected, 0.2mL of cell suspension is injected subcutaneously at the root of thigh in a clean bench, a local obvious skin dune is injected, the nude mouse is normally raised for 14 days after inoculation, the major diameter (a) and the minor diameter (b) of a tumor nodule are measured by a vernier caliper, and the approximate volume of the tumor is estimated according to the formula V which is ab 2/2.

2.2 screening of mouse Colon cancer subcutaneous transplantation high metastasis model [ screening and establishment of mouse Colon cancer high metastasis model in vivo ]

NOD-SCID mice are selected for in vivo primary screening of tumors. Mice were first resected for subcutaneous tumors to extend tumor-bearing life. In the dying state, lung metastases with obvious visibility of the fleshy eyes are transplanted to the next generation of NOD-SCID mice for subcutaneous amplification. After 3 times of repeated subcutaneousness implantation of mice → lung metastasis → subcutaneous amplification → lung metastasis screening, the lung metastasis focus is transplanted to the subcutaneous part of BALB/c nude mice with normal immune system, and the tumor in-vivo screening of BALB/c nude mice is continued (the method steps are the same as that of NOD-SCID mice, the tumor in-vivo primary screening is carried out, and the screening is carried out for 3 times).

2.3 subcutaneous transplantation tumor in situ inoculation

Normally raising nude mice with high metastasis model for 14 days after the inoculation of the nude mice with colon cancer subcutaneous transplantation, cutting tumor body tissues of the nude mice with tumor formation into small blocks with the diameter of about 0.1cm on the 15 th day, performing inhalation anesthesia by diethyl ether before 60 nude mice, sterilizing the skin conventionally, opening the abdominal cavity, pulling out intestinal segments, using an injection needle to cut a serous layer, using biological adhesive to adhere the tumor body and the intestinal wall, using normal saline to press for 2 minutes, returning back to the intestinal tract, suturing peritoneum (containing muscular layer) and the skin, and using antibiotics to prevent infection after the operation.

And (4) performing postoperative group cage-division feeding (SPF grade) and observation, dissecting and taking materials when the tumor is in an moribund state, recording and observing the tumor infiltration and metastasis conditions, observing the inoculated tumor and measuring the weight of the tumor body.

2.4 grouping and administration methods

Nude mice were divided into six groups of group a (model group), group B (5-fluorouracil group), group C (oxaliplatin group), group D (5-fluorouracil + oxaliplatin group), group E (chinese herbal medicine + 5-fluorouracil + oxaliplatin group), and group F (glutamine + 5-fluorouracil + oxaliplatin group) by body weight 24h after in situ inoculation (day 16), 10 mice were housed in each group, and 5 mice were housed in each cage with free diet.

Beginning on the third day of the orthotopic transplantation operation, the administration is started according to the injection dosage of 0.1ml/10g, the dose of the chemotherapy drug is converted according to the body surface area of an adult, and the calculation formula of the drug dose is converted: dB dA × (RB/RA) × (WA/WB) 1/3. Group B, the intraperitoneal injection is given every other day according to the dose of 25mg/kg, and 5 times of medication are given in total. Group C: the preparation is administered by intraperitoneal injection at 6.6mg/kg every other day for 5 times. Group D: 6.6mg/kg of oxaliplatin is intraperitoneally injected every other day, and 25mg/kg of 5-fluorouracil is intraperitoneally injected every other day, and the medicines are taken for 5 times. Gavage was continued from day 17 (the second day after orthotopic transplantation surgery) to day 38 for 21 days, and gavage was given to four groups AB C D with physiological saline. The group E and the group F are administered with Chinese medicinal materials for intragastric administration and 2g/kg of glutamine for intraperitoneal injection. Water was only provided to fast on day 39. On day 40, the cervical vertebra was sacrificed, blood was taken from the eyeball, and ileum tissue and intestinal tumor tissue were dissected out.

3. Detecting the index

3.1 Western Blot to detect PI3K, AKT protein.

3.2 Real-Time PCR detection of AKT, PI3K mRNA expression in the intestinal mucosa.

3.3 ileal epithelial injury index assessment.

3.4 immunohistochemistry (ileum): AKT, PI3K expression in intestinal mucosa.

3.5 HE multiple sampling (10 points) the lung and liver metastasis counts were analyzed.

4. Results

4.1 weight of nude mice (see FIG. 1) and weight of intestinal tumor (see FIGS. 2 and 3, Table 1)

TABLE 1 weight comparison of nude mice and intestinal tumors in each group

Note: compared to group a (model group), # p < 0.05; p <0.05 compared to group E

4.2 WB test intestinal tissue protein expression results (see FIG. 4)

The intestinal tissue is cracked by RIPA lysate, protein quantification is carried out according to the kit step method, primary antibody and secondary antibody are added for incubation, washing is carried out, and finally semi-quantitative analysis is carried out.

4.3 PCR detection of PI3K, AKT mRNA expression in intestinal tumor tissue (see FIGS. 5-8, Table 3)

The upstream and downstream primer sequences were designed by Shanghai Kilton (see Table 2). The procedures of the kit instructions were followed.

TABLE 2 mouse Up and Down primers

TABLE 3 comparison of PI3K, AKT expression in various groups of intestinal tumor tissues

Note: p <0.05 compared to group a (model group); p <0.05 compared to group E (body resistance strengthening and spleen invigorating plus 5-fluorouracil plus oxaliplatin).

4.4 ileal epithelial Damage index assessment (see FIG. 9, Table 4)

The extent of ileal mucosal injury was assessed by determining the ileal epithelial injury index according to the Chiu's 6 scale scoring method: level 0 is normal; grade 1 is the widening of the subepithelial space at the top of the villus; grade 2, the gap under the epithelium at the top of the villus is further enlarged, and the epithelium at the top of the villus is lifted and peeled from the inherent membrane; grade 3, the villous epithelium is broken off; grade 4, complete desquamation of epithelium, only inherent membrane; grade 5 is the disintegration of the lamina propria, with bleeding and ulceration. The ileal intestinal villus length, width and crypt depth were measured using a microscope scale, and 10 fields were observed at random by 3 pathologists per section, averaged, and the images were read and photographed under a microscope.

TABLE 4 comparison of ileal epithelium in nude mice of each group

Note: compared to group a (model group), # p < 0.05; p >0.05 compared to group E;

4.5 pulmonary and hepatic metastasis counts (see FIG. 10)

The lung and liver tissues are dehydrated conventionally, embedded in paraffin, and subjected to histopathological examination after section HE staining. The lung and liver metastases were calculated using a multi-point sampling and sectioning procedure (10 points per tissue).

5. Statistical treatment:

statistical analysis software used SPSS 20.0. If the measured data are in accordance with normal distribution and the variance is uniform, the data are represented by +/-meter, variance analysis is adopted for comparison among groups, and variance analysis designed by repeated measurement is adopted for repeated measured data; if the distribution is not in accordance with the normal distribution and the variance is irregular, non-parametric test is used, and if the p is less than 0.05, the difference has statistical significance.

6. Discussion:

the preparation can inhibit tumor cell proliferation, reduce intestinal inflammatory cytokine expression, and relieve intestinal injury caused by chemotherapy, and can effectively control tumor size and effectively reduce expression of PI3K and AKT by combined chemotherapy. The formula is proved to protect intestinal barrier, regulate PI3K/AKT expression in intestinal tumor microenvironment and inhibit metastasis.

Example 2 the efficacy of the present invention is compared to the efficacy of the enteral decoction formulation

1. Animals: 20 BALB/c nude mice 4-5 weeks old, 20-24 g in body mass, purchased from Shanghai Si Laike laboratory animals Co., Ltd, Chinese academy, raised in SPF-level environment, and certified animal number: SYXK (Shanghai) 2014-0008.

2. The method comprises the following steps:

2.1 cell culture and subcutaneous transplantation of tumors

Method of same animal experiment 1

2.3 subcutaneous transplantation tumor in situ inoculation

Method of same animal experiment 1

2.4 grouping and administration methods

Nude mice were divided into a group a (traditional Chinese medicine formulation + 5-fluorouracil + oxaliplatin group) and a group B (intestine easy decoction + 5-fluorouracil + oxaliplatin group) by weight 24h (day 16) after in situ inoculation, 10 mice were fed in cages each with 5 mice per cage, and were fed with free diet. The intestine-benefiting decoction formula comprises: 10 parts of codonopsis pilosula, 15 parts of poria cocos, 10 parts of bighead atractylodes rhizome, 15 parts of rhizoma smilacis glabrae, 15 parts of chinaroot greenbrier, 12 parts of akebia fruit, 12 parts of endothelium corneum gigeriae galli and 9 parts of pericarpium citri reticulatae.

A. Group B: 6.6mg/kg of oxaliplatin is intraperitoneally injected every other day, and 25mg/kg of 5-fluorouracil is intraperitoneally injected every other day, and the medicines are taken for 5 times. The two groups AB are administered with the Chinese medicinal prescription and the intestine-benefiting decoction respectively. Water was only provided to fast on day 39. On day 40, the cervical spine was sacrificed and intestinal tumor tissue was dissected out.

3. Results

3.1 weight of nude mice and weight of intestinal tumor in each group (see Table 5)

TABLE 5 weight comparison of nude mice and intestinal tumors in each group

Note: compared to group B (model group), # p < 0.05.

Compared with the intestine-benefiting decoction formula, the weight of the mice and the tumor-removing weight are both higher than those of the intestine-benefiting decoction group, the difference has statistical significance (P is less than 0.05), and compared with the tumor weight of the mice, the model group is higher than that of the A group (P is less than 0.05). The prescription can regulate the immune function of tumor-bearing animals, and has better curative effect on increasing the weight of mice and inhibiting tumor bodies than the intestine-benefiting decoction prescription.

Example 3 clinical trial study

1. Case selection:

60 patients who were diagnosed as colorectal cancer at the time of admission in Xinhua hospital affiliated to Shanghai university of transportation during 2017, month 06 to month 03 in 2018, and patients who were diagnosed as colorectal cancer at my hospital (the combined Hospital and Western medicine hospital affiliated to Shanghai university of medicine) were selected, and chemotherapy was performed on all patients during this period.

The Western diagnosis was made according to the diagnostic criteria of the clinical oncology society of China "CSCO diagnostic guidelines for colorectal cancer" (2016 edition). TNM pathological staging (pTNM) AJCC/UICC 7 th edition [8], see in detail "staging of colorectal cancer 2.4".

The syndrome of traditional Chinese medicine is diagnosed by researchers qualified by doctors in the department according to the clinical research guiding principle of traditional Chinese medicine for treating spleen deficiency syndrome [9] whether the diagnosis meets the diagnosis standard of the spleen deficiency syndrome or whether the two doctors have the same syndrome differentiation conclusion. In clinical manifestations, more than or at least 2 primary symptoms of spleen deficiency are required, or 1 primary symptom of spleen deficiency is satisfied with 2 secondary symptoms of spleen deficiency, wherein the tongue pulse must meet the diagnosis standard of spleen deficiency.

1.1 case inclusion criteria

(1) The diagnosis standard after colorectal cancer surgery is met, and pathological examination is definitely diagnosed as stages III and IV;

(2) postoperative adjuvant chemotherapy indications;

(3) age 18-75 years old, male and female;

(4) KPS score is more than or equal to 60 points, and life time is more than or equal to 3 months;

(5) meets the diagnosis standard of spleen deficiency syndrome.

(6) The subject was subjected to the clinical trial.

1.2 exclusion criteria:

(1) those that do not meet the inclusion criteria.

(2) Serious primary diseases such as cardiovascular and cerebrovascular diseases, or mental diseases;

(3) suffering from other severe complications, such as pancreatitis, burns, wounds, etc.;

(4) 1 month before the group, the history of the antibiotics is available;

(5) patients suffering from or associated with other intestinal disorders, such as enteritis, intestinal obstruction, etc., in addition to the cancer;

(6) any unfavorable selection was considered in the study.

3. The medicine taking method comprises the following steps:

the patients meeting the included standard of 60 middle-stage colorectal cancer chemotherapy patients are randomly divided into 30 treatment groups and 30 control groups, the treatment groups are treated by the drug preparation combined chemotherapy, and the control groups are treated by simple chemotherapy. All patients underwent chemotherapy for more than 2 courses.

The chemotherapy scheme comprises a common chemotherapy scheme for colorectal cancer, and the chemotherapy scheme is established according to the NCCN guideline according to the individual condition of a patient. 21 days is 1 course of treatment, and the treatment course is more than 2.

The treatment groups take the medicine preparation orally at the same time, 2 times a day, 1 bag each time. 21 days is a treatment course, and two treatment courses are taken. 4. Evaluation indexes are as follows: traditional Chinese medicine symptom integral and KPS score, chemotherapy toxic and side effect, adopt two sets of patients to treat the front and back periphery serum one respectively simultaneously, adopt ELISA method to detect two sets of patients periphery blood intestinal barrier function, including mechanical intestinal barrier index: DAO, D-LA, IFABP, ET; immune intestinal barrier indexes, sIgA, IL-2, TNF-beta, TGF-beta, and IFN-gamma.

4.1 Chinese medicine syndrome integration

The clinical syndrome curative effect judgment standard.

The effect is shown: the total integral value of clinical symptoms after treatment is reduced by more than or equal to 70 percent compared with that before treatment;

the method has the following advantages: the total integral value of clinical symptoms after treatment is reduced by less than 70 percent and more than or equal to 30 percent than before treatment;

and (4) invalidation: the total integral value of clinical symptoms after treatment is reduced by less than 30 percent compared with that before treatment.

Note: the calculation formula is as follows: [ (pre-treatment integral-post-treatment integral) ÷ pre-treatment integral ] × 100%.

4.2 quality of life

4.2.1 behaviour KPS score: and respectively recording the KPS scores once before and after grouping by referring to a KPS score scale.

TABLE 6 KPS Scoring Scale

The evaluation criteria for efficacy on the Ka-score are as follows:

the improvement is as follows: the integral value after treatment is increased by more than or equal to 10 points compared with the integral value before treatment;

and (3) stabilizing: (ii) a score increase or decrease of less than 10 points after treatment compared to pre-treatment score;

and (3) descending: the score after treatment is reduced by more than or equal to 10 points compared with the score before treatment.

4.2.2 scoring method (rough score RS) in each field of QLQ-C30, dividing QLQ-C30 scale into 15 fields according to application and scoring method of QLQ-C30 of Qiu color peak and Zhao Jun, and calculating score of each field:

TABLE 7 QLQ-C30 statistics Table

Normalized score (standard score, ss): in the QLQ-C30 scale, the entries are all reversed entries (the larger the value, the worse the quality of life). The scoring rules clearly specify that higher scores for the functional domain and the general health domain indicate better functional status and quality of life; a higher score for a symptom domain indicates more symptoms or problems (worse quality of life). Therefore, the scaling of the computing function domain is also changed in direction. The calculation is as follows (where R is the total distance of the scores of the respective domains or items).

The functional field is as follows: SS ═ 1- (RS-1)/R ═ 100

Symptom field and general health field: SS ═ R [ (RS-1)/R ]. 100

QLQ-C30 score evaluation criteria for efficacy:

the effect is shown: the integral value after treatment is increased by more than or equal to 20 points compared with the integral value before treatment;

the method has the following advantages: the score increases more than 10 points and less than 20 points after treatment compared with before treatment;

no change: the score increased or decreased by 10 points after treatment compared to before treatment.

Deterioration: the score after treatment is reduced by more than or equal to 10 points compared with the score before treatment

4.3 chemotherapy toxic and side effects: before and after each chemotherapy, the routine examination of blood and the functions of liver and kidney are performed, and the examination times are increased according to actual needs.

Judging according to WHO antitumor drug acute and subacute toxic reaction graduation standard, dividing into 0 degree, I degree, II degree, III degree, IV degree, and treating with colony cell stimulating factor if bone marrow suppression above II degree occurs during chemotherapy.

TABLE 8 graduation Standard for acute and subacute toxicity reactions of antitumor Agents (WHO)

Note: n: normal value

4.4 intestinal barrier function: intestinal barrier function was measured and recorded before treatment and after observation for all patients in the group.

Including the mechanical intestinal barrier indexes: DAO, D-LA, IFABP, ET; immune intestinal barrier index: sIgA), IL-2, TNF-beta, TGF-beta, IFN-gamma. All the indexes are detected by Elisa at scientific and technological experiment center of Shanghai medical university.

5. Statistical analysis

Using SPSS 22.0 software to carry out statistics, firstly adopting a normality test for metering data, and carrying out a bilateral test by using a statistical method of a t test if the metering data accords with the normally distributed metering data; and carrying out bilateral test on the metering data and the grade data which do not conform to normal distribution by adopting a statistical method of rank sum test. Differences were considered statistically significant if p <0.05 or p < 0.001.

6. The results show that:

6.160 comparison of Chinese medicine syndrome before and after treatment (see Table 9)

Table 9 comparative units of clinical efficacy of traditional chinese medical symptoms after treatment of two groups of cases: example (%)

Note: the Mann-Whitney U test shows that the difference is statistically significant when Z is-4.178 and P is less than 0.001

6.2 comparison of quality of Life between control and treatment groups

6.2.1 comparison of KPS scores before and after treatment with the control group (see Table 10)

Table 10 two groups of case quality of life comparison units: example (%)

Note: the Mann-Whitney U test shows that the difference is statistically significant, Z is-2.881, P is less than 0.05

6.2.2 comparison of QLQ-C30 scores between control and treatment before and after treatment

6.2.2.1 comparison of Life function between control and treatment patients (see Table 11)

TABLE 11 comparison of QLQ-30 score efficacy before and after treatment of two groups of cases

Note: by Mann-Whitney U test, two groups are compared, P is less than 0.05, and the difference has statistical significance.

In the symptom domain and the aspects of breathlessness, insomnia, loss of appetite, constipation and diarrhea, the score of tiredness, pain, breathlessness, insomnia, loss of appetite, constipation and diarrhea of the treatment group is reduced after the treatment compared with that before the treatment, and the score of nausea and vomiting is unchanged, wherein the difference between tiredness and loss of appetite is statistically significant (P is less than 0.05) compared with that before the treatment, and the difference between tiredness, nausea and vomiting, loss of appetite and diarrhea is statistically different (P is less than 0.05) compared with that of the control group. The pain and constipation scores of the control group decreased after treatment compared to before treatment, while the scores for tiredness, nausea and vomiting, breathlessness, insomnia, loss of appetite, diarrhea all increased earlier, with the values for tiredness, nausea and vomiting and diarrhea statistically significant compared to the differences before treatment (P < 0.05). The prescription can improve symptoms of fatigue, nausea and vomiting, breathlessness, insomnia, loss of appetite and diarrhea caused by chemotherapy, wherein the improvement effect on the fatigue, the nausea and vomiting, the loss of appetite and the diarrhea is particularly obvious.

6.3 comparison of the toxicity and adverse effects of chemotherapy in the control and treatment groups (see Table 12)

TABLE 12 comparison of blood routine and liver and kidney function indexes before chemotherapy in two groups of cases (unit: case)

Note: by Mann-Whitney U test, white blood cells: z ═ 1.472, P ═ 0.141; neutrophils: z ═ 2.817, P ═ 0.005; hemoglobin: z ═ 2.220, P ═ 0.026; platelets: z ═ 1.762, P ═ 0.078; glutamic-oxalacetic transaminase: z ═ 0.399, P ═ 0.690; glutamic-pyruvic transaminase: z ═ 0.447, P ═ 0.655; creatinine: z ═ 1, P ═ 0.317; urea nitrogen: z is 0, P is 1.000; nausea and vomiting: -2.201, 0.028 for Z; diarrhea: z is-2.246 and P is 0.025. P is less than 0.05, and the difference has statistical significance

Statistical analysis shows that the difference between the two groups in terms of neutrophilic granulocyte, hemoglobin, nausea, vomiting and diarrhea has statistical significance (P is less than 0.05). The prescription has good safety, can improve the reduction of neutrophils and hemoglobin caused by chemotherapy, and can improve the digestive tract toxic and side effects of nausea, vomiting and diarrhea caused by chemotherapy.

6.4 comparison of peripheral serum indices before and after treatment in two groups of patients

6.4.1 intestinal tract mechanical Barrier function (see Table 13)

TABLE 13 serum levels of DAO, D-LA, IFABP, ET in two groups of patients before and after treatment

Note: (1) comparison of treatment groups with control groups: through independent t-test analysis, compared with a control group, P is less than 0.001, and the difference has statistical significance; (2) comparison before and after treatment: through the test of a matched sample t, compared with the sample before treatment, the # P is less than 0.001, the delta P is less than 0.05, and the difference has statistical significance.

Compared with the control group, the D-LA content in the treatment group is obviously reduced, and the difference has statistical significance (P is less than 0.001).

Compared with the treatment group before and after treatment, the D-LA treatment is obviously reduced compared with the treatment group before and after treatment, the difference between the D-LA treatment and the treatment group before and after treatment has obvious statistical significance (P is less than 0.001), and the DAO, IFABP and ET treatment are all increased compared with the treatment group before treatment but have no statistical significance (P is more than 0.05).

Compared with the control group before and after treatment, the DAO, D-LA, IFABP and ET are all increased compared with the control group before treatment, wherein the D-LA values are statistically different before and after treatment (P is less than 0.05).

6.4.2 intestinal immune barrier function (see Table 14)

TABLE 14 serum levels of sIgA, IL-2, TNF-beta, TGF-beta, IFN-gamma for two groups of patients before and after treatment

Note: (1) treatment and control groups: through independent t-test analysis, compared with a control group, P is greater than 0.05, and the difference has no statistical significance; (2) comparison before and after treatment: after the matched sample t test, compared with the sample before treatment, the P is more than 0.05, and the difference has no statistical significance.

Compared with the control group, after treatment, sIgA, IL-2 and IFN-gamma tend to be increased, and TNF-beta and TGF-beta serum contents tend to be reduced, but the comparison has no statistical significance (P is more than 0.05).

Compared with the treatment group before and after treatment, sIgA and IL-2 show an increasing trend, and the difference between the front and the back has no significant statistical significance (p is more than 0.05), TNF-beta, TGF-beta and IFN-gamma show a decreasing trend after treatment, but have no statistical significance (p is more than 0.05).

Compared with the control group before and after treatment, IFN-gamma is in a decreasing trend, sIgA, IL-2, TNF-beta and TGF-beta are in an increasing trend, wherein the increasing trend of the sIgA index is not obvious as that of the treatment group, and all indexes before and after the treatment of the control group have no statistical difference (P is more than 0.05).

7. And (4) conclusion:

after chemotherapy, the content of D-LA in serum of patients with middle and late colorectal cancer and spleen deficiency syndrome is obviously increased, and the serum content of sIgA, IL-2 and IFN-gamma is reduced, so that the formula can obviously reduce the content of D-LA and increase the indexes of sIgA, IL-2 and IFN-gamma. The suggestion is that the traditional Chinese medicine composition can inhibit intestinal mucosa permeability increase and intestinal inflammatory reaction caused by chemotherapy to a certain extent, improve the mechanical barrier function of the intestinal tract, play an immune protection role in the intestinal tract and improve the immune barrier function of the intestinal tract. The medicinal preparation can improve Chinese medicine symptom integration by combining chemotherapy treatment; the pharmaceutical preparation can improve KPS scores of patient behavior by combining chemotherapy; by comparing various indexes in QLQ-C30 scales of two groups of patients, the pharmaceutical preparation can improve the body function of the patients, improve the role function of the patients, relieve the bad mood of the patients, and improve the symptoms of fatigue, nausea and vomiting, breathlessness, insomnia, loss of appetite and diarrhea caused by chemotherapy in various indexes of the life functions of the patients, wherein the improvement effect on the fatigue, nausea and vomiting, loss of appetite and diarrhea is particularly obvious. Improving the overall health status of the patient; the pharmaceutical preparation can not increase the toxic and side effects of chemotherapy when being used by patients after colorectal cancer operation in combination with chemotherapy, has good safety, has no obvious influence on the economic burden of the patients after the chemotherapy, and is equivalent to the simple chemotherapy.

Preparation examples

The preparation method of the compound Chinese medicine preparation of the present invention is further illustrated by the following examples.

Preparation of the Chinese medicinal compound granule

(1) Weighing 30g of astragalus, 10g of codonopsis pilosula, 10g of poria cocos, 10g of bighead atractylodes rhizome, 10g of rhizoma smilacis glabrae, 10g of smilax china, 10g of fiveleaf akebia fruit, 5g of endothelium corneum gigeriae galli and 10g of Chinese actinidia root for later use; (2) taking half of the raw materials (15 g of astragalus, 5g of codonopsis pilosula, 5g of poria cocos, 5g of bighead atractylodes rhizome, 5g of glabrous greenbrier rhizome, 5g of chinaroot greenbrier rhizome, 5g of fiveleaf akebia fruit, 2.5g of chicken gizzard membrane and 5g of Chinese actinidia root) of the traditional Chinese medicines, adding 8 times of water, soaking for half an hour, decocting for 45 minutes, adding 8 times of water again, decocting for 30 minutes, filtering, combining decoction, concentrating under reduced pressure, drying under reduced pressure to obtain dry extract, and crushing into extract fine powder (passing through 80 meshes.

(3) Taking the other half amount of the raw materials (15 g of astragalus, 5g of codonopsis pilosula, 5g of poria cocos, 5g of bighead atractylodes rhizome, 5g of glabrous greenbrier rhizome, 5g of chinaroot greenbrier rhizome, 5g of fiveleaf akebia fruit, 2.5g of chicken's gizzard-membrane and 5g of Chinese actinidia root) of the traditional Chinese medicines, crushing the raw materials into 80 meshes, sterilizing the crushed raw materials by using Co60 (the irradiation dose is 8k), mixing the sterilized fine powder with the extract, adding mannitol, lactose, sucrose powder, aspartame and menthol, uniformly mixing the mixture and then bonding the mixture by using 70% ethanol to prepare.

Preparation of the Chinese medicinal compound tablet

(1) Weighing 45g of astragalus mongholicus, 30g of codonopsis pilosula, 20g of poria cocos, 20g of bighead atractylodes rhizome, 15g of rhizoma smilacis glabrae, 30g of smilax china, 20g of fiveleaf akebia fruit, 20g of endothelium corneum gigeriae galli and 30g of Chinese actinidia root for later use;

(2) taking half of the raw materials of the traditional Chinese medicines (namely 22.5g of astragalus, 15g of codonopsis pilosula, 10g of poria cocos, 10g of bighead atractylodes rhizome, 7.5g of glabrous greenbrier rhizome, 15g of chinaroot greenbrier rhizome, 10g of fiveleaf akebia fruit, 10g of chicken gizzard membrane and 15g of Chinese actinidia root), adding 5 times of water, soaking for half an hour, decocting for 45 minutes, adding 3 times of water, decocting for 30 minutes, filtering, combining the decoction, concentrating under reduced pressure, drying under reduced pressure to obtain dry extract, and crushing into extract fine powder (passing through a 80-.

(3) Taking the other half amount of the raw materials (22.5 g of astragalus, 15g of codonopsis pilosula, 10g of poria cocos, 10g of bighead atractylodes rhizome, 7.5g of glabrous greenbrier rhizome, 15g of chinaroot greenbrier rhizome, 10g of fiveleaf akebia fruit, 10g of chicken gizzard membrane and 15g of Chinese actinidia root) of the traditional Chinese medicines, crushing the raw materials into 80 meshes, sterilizing the powder by using Co60 (irradiation dose is 8k), mixing the powder with the extract fine powder, adding mannitol, lactose, sucrose powder, aspartame and menthol, uniformly mixing, bonding by using 70% ethanol, drying, adding a proper amount of magnesium stearate, uniformly mixing, and pressing to obtain the traditional Chinese.

Preparation of the traditional Chinese medicine compound capsule of the invention

(1) Weighing 30g of astragalus, 20g of codonopsis pilosula, 15g of poria cocos, 15g of bighead atractylodes rhizome, 10g of rhizoma smilacis glabrae, 15g of smilax china, 15g of fiveleaf akebia fruit, 12g of endothelium corneum gigeriae galli and 15g of Chinese actinidia root for later use;

(2) taking half of the raw materials of the traditional Chinese medicines (namely 15g of astragalus, 10g of codonopsis pilosula, 7.5g of poria cocos, 7.5g of bighead atractylodes rhizome, 5g of glabrous greenbrier rhizome, 7.5g of chinaroot greenbrier rhizome, 7.5g of fiveleaf akebia fruit, 6g of chicken's gizzard-membrane and 7.5g of Chinese actinidia root), adding 3 times of water, soaking for half an hour, decocting for 45 minutes, adding 8 times of water, decocting for 30 minutes, filtering, combining decoction, concentrating under reduced pressure, drying under reduced pressure to obtain dry extract, and crushing into extract fine powder (passing through 80.

(3) Taking the other half amount of the raw materials (namely 15g of astragalus, 10g of codonopsis pilosula, 7.5g of poria cocos, 7.5g of bighead atractylodes rhizome, 5g of glabrous greenbrier rhizome, 7.5g of chinaroot greenbrier rhizome, 7.5g of fiveleaf akebia fruit, 6g of chicken's gizzard-membrane and 7.5g of Chinese actinidia root), crushing and sieving by 80 meshes, sterilizing by Co60 (irradiation dose is 8k), mixing with the extract fine powder, adding mannitol, lactose, sucrose powder, aspartame and menthol, uniformly mixing, bonding by 70% ethanol, drying, crushing, adding a proper amount of starch, uniformly mixing, and encapsulating to prepare the hard capsule.

It will be understood by those of ordinary skill in the art that the foregoing embodiments are specific examples for carrying out the invention, and that various changes in form and details may be made therein without departing from the spirit and scope of the invention in practice.

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Claims

1. A traditional Chinese medicine compound preparation for improving intestinal barrier function and regulating body immunity of tumor patients is characterized by being prepared from the following traditional Chinese medicine raw materials in parts by weight: 30-45 parts of astragalus membranaceus, 10-30 parts of codonopsis pilosula, 10-20 parts of poria cocos, 10-20 parts of bighead atractylodes rhizome, 10-15 parts of rhizoma smilacis glabrae, 10-30 parts of chinaroot greenbrier, 10-20 parts of fiveleaf akebia fruit, 5-20 parts of endothelium corneum gigeriae galli and 10-30 parts of Chinese actinidia root.

2. The compound traditional Chinese medicine preparation of claim 1, which is prepared from the following traditional Chinese medicine raw materials in parts by weight: 30 parts of astragalus membranaceus, 20 parts of codonopsis pilosula, 15 parts of poria cocos, 15 parts of bighead atractylodes rhizome, 10 parts of rhizoma smilacis glabrae, 15 parts of smilax china, 15 parts of fiveleaf akebia fruit, 12 parts of endothelium corneum gigeriae galli and 15 parts of Chinese actinidia root.

3. A method for preparing the compound traditional Chinese medicine preparation of claim 1, comprising the following steps:

(1) weighing the traditional Chinese medicine raw materials according to the prescription amount for later use;

(3) taking the other half amount of the raw materials of the traditional Chinese medicines in the step (1), crushing, sterilizing, mixing with the extract fine powder prepared in the step (2), further adding pharmaceutically acceptable auxiliary materials, and preparing the traditional Chinese medicine compound preparation by a conventional traditional Chinese medicine preparation method.

4. The preparation method according to claim 3, wherein 3 to 8 times of water is added during the first decoction in the step (2), and after the water is soaked for half an hour, the water is decocted for 45 minutes; and 3-8 times of water is added during the second decoction, and the decoction is carried out for 30 minutes.

5. The method of claim 3, wherein the sterilization method in the step (3) is Co60 irradiation sterilization.