CN115192617A - Application of Babaodan in preparation of medicine for preventing relapse of early liver cancer after operation - Google Patents
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Abstract
The invention provides an application of Babaodan in preparing a medicine for preventing postoperative recurrence of liver cancer. Pharmacological experiments find that the Babaodan has obvious effect of preventing postoperative recurrence of the liver cancer. Therefore, the eight-treasure pill can be used for preparing the medicine for preventing the postoperative recurrence of the liver cancer and has good development and application prospects.
Description
The application is a divisional application of an invention patent application with the application number of 2016110656266, the application date of 2016, 11 and 28, and the title of the invention is "application of eight-treasure pill in preparation of medicine for preventing postoperative recurrence of early stage hepatocarcinoma".
Technical Field
The invention belongs to the field of new drug indications, and particularly relates to application of Babaodan in preparing a drug for preventing postoperative recurrence of liver cancer.
Background
Liver cancer is one of common malignant tumors in China, and has high mortality rate. The best treatment means for the early liver cancer is radical surgical resection at present. However, patients with liver cancer are easy to relapse and transfer after operation, early liver cancer relapse concentrates in two years after operation, and the relapse rate exceeds 50 percent.
The tumor microenvironment which is the hotspot of the current research plays an indispensable role in the recurrence and metastasis of liver cancer. A great deal of research shows that the tumor micro-environment has a non-negligible effect on the occurrence and development of tumors. According to the "seed and soil" theory, the microenvironment of the target organ for recurrent metastasis is one of the determining factors influencing whether recurrent metastasis occurs. It is now recognized that metastasis is a process of interaction between tumor cells and the host microenvironment, which can affect the biological properties of the tumor and promote or inhibit colonization of the target organ by tumor cells. If the inflammatory state of the target organ favorable for metastasis formation can be weakened, the occurrence of metastasis can be reduced. However, the metastasis and recurrence after the liver cancer operation mostly occur in the residual liver after the operation, namely the metastasis and recurrence target organ is the liver, so the microenvironment of the liver, especially the inflammatory immune microenvironment, can directly influence the prognosis of the patient after the liver cancer operation. Postoperative recurrence of liver cancer is due to the selective amplification of disseminated neoplastic cells by the residual liver after surgery, forming recurrent foci, or the selective amplification of newly mutated cells, forming new foci of liver cancer. The inflammatory state of the residual liver after liver cancer surgery and the regenerative capacity of the residual liver are inversely related, and the heavier the inflammatory reaction is, the poorer the regenerative capacity of the residual liver is, the more easily a tumor recurrence focus is formed. The precancerous liver tissue well represents the state of the residual liver after operation. In conclusion, treatment of liver inflammation helps to inhibit tumor recurrence and metastasis.
The eight-treasure pill is prepared with ox gallstone, snake gall, antelope horn, pearl, notoginseng, musk and other famous and precious medicinal materials, and contains several effective components, such as taurine, musk ketone, taurocholic acid, saponin and flavone. The monarch drug natural bezoar is cool in nature and bitter in taste, calms the liver and relieves convulsion, clears away heat and toxic materials, mainly contains taurine as an effective component, contains bilirubin and bile acid, can dilate capillary vessels and relax smooth muscles of biliary tracts, and has the effects of benefiting the liver and benefiting the gallbladder and promoting the recovery of hepatitis; the snake gall as a ministerial drug is cool in nature and slightly bitter in taste, has the effects of clearing away heat and toxic materials, relieving pain and improving eyesight, contains more than 12 kinds of bile acids, mainly contains ursodeoxycholic acid, and has the effect of dissolving bile salt calculus; the adjuvant drug pearl has cold nature and salty taste, has the effects of relieving palpitation and arresting convulsion, astringing and promoting granulation, and clearing liver and removing nebula; the antelope horn as adjuvant drug has the efficacies of antipyresis, antivirus and immunity enhancement; the natural musk of the guiding drug has warm and pungent properties, promotes blood circulation, removes blood stasis, reduces swelling and relieves pain, and the main components are musk ketone and polypeptide which can relieve hepatitis symptoms; the active component of the medicinal notoginseng is the total glycoside of notoginseng, which has the functions of eliminating free radical, activating mercaptoenzyme system, protecting liver and resisting oxidation.
Pharmacological research finds that the eight-treasure pill has strong anti-inflammatory and analgesic, liver and gallbladder protection equivalent effects, is clinically used for treating various damp-heat syndromes, including jaundice and digestive tract symptoms caused by damp-heat, and liver protection treatment, but reports on the aspect of preparing a medicine for preventing early liver cancer postoperative recurrence are not seen yet.
Disclosure of Invention
The invention aims to provide the application of Babaodan in preparing a medicine for preventing postoperative recurrence of early liver cancer according to the current situation that the Chinese medicinal composition has the effect of preventing postoperative recurrence of early liver cancer.
The BABAODAN Capsule is prepared from Xiamen Chinese medicinal plant Limited company, and has Chinese medicine standard Z10940006, and comprises calculus bovis, fel Serpentis, cornu Saigae Tataricae, margarita powder, notoginseng radix, and Moschus as main ingredients.
The invention relates to an application of Babaodan in preparing a medicine for preventing relapse after early liver cancer surgery, which belongs to the first disclosure.
Has the advantages that: the invention has definite and obvious curative effect through preclinical verification. Pharmacological experiments find that the medicine has obvious effect of preventing early liver cancer postoperative recurrence. Therefore, the eight-treasure pill can be used for preparing the medicine for preventing the early-stage liver cancer postoperative recurrence and has good development and application prospects.
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FIG. 1 is a graph showing the intervention effect of Babaodan on liver cancer occurrence in a rat Diethylnitrosamine (DEN) induced primary liver cancer model, liver function detection and staining of pathological sections; FIG. 1A shows gross specimens of rat liver after various treatments (black arrows represent tumor tissue); FIG. 1B shows the incidence of liver cancer in each group; figure 1C shows the mean levels of serum albumin, glutamic-pyruvic transaminase and glutamic-oxaloacetic transaminase of each group of rats (student-t test,. P < 0.01); FIG. 1D shows HE staining of liver tissue of each group;
FIG. 2 is a diagram showing the development of hepatoblastoma by Lipopolysaccharide (LPS) treatment of six groups of rats and the size and number of liver cancers of the rats treated with babaodan at different stages;
FIG. 3 is a pathological section of liver cancer tissue and tissues adjacent to the cancer after rat dissection; the cancer tissue was stained by hematoxylin-eosin (HE) staining (upper panel), and the cancer-adjacent tissue was stained by HE (lower panel) (microscope magnification is 200-fold).
Detailed Description
The following examples serve to illustrate the invention.
SD rats were purchased from Shanghai Stark laboratory animals Co., ltd and diethylnitrosamines were purchased from Sigma Co., ltd.
Example 1
Study on the effect of Babaodan on the development of primary liver cancer in rats caused by diethylnitrosamine (DEN water, DEN volume concentration of 1.
After 30 SD rats were adaptively fed for one week, they were randomly divided into 4 groups, i.e., a normal group (Ctrl group, 5), a model group (DEN group, 10), a test drug eight-treasure lead group (BBD group, 5), and a liver cancer model administration group (DEN + BBD group, 10).
The 4 groups of SD rats were treated separately as follows: (1) normal group: feeding purified water; (2) model group: feeding purified water containing diethylnitrosamine (DEN volume concentration of 0.1 g/L); (3) the tested medicine Babaodan group: drinking purified water, and performing intragastric administration twice a week, wherein the administration mode is that the powder of the Babaodan in the Babaodan capsule is dissolved in the purified water and administered intragastric administration, and the administration dosage of each time is 0.5g/kg calculated by the powder of the Babaodan; (4) administration group of liver cancer model: feeding purified water containing diethylnitrosamine (wherein DEN volume concentration is 1 10000) and performing intragastric administration twice a week, wherein the administration mode is that the powder of the eight-treasure pill in the eight-treasure pill capsule is dissolved in the purified water and is subjected to intragastric administration, and the administration dose is 0.5g/kg in terms of the powder of the eight-treasure pill every time, and feeding for 12 weeks. 12 weeks after the start of the experimental treatment, the patients were sacrificed under anesthesia, and the liver and blood were taken for further analysis.
The experimental results are as follows:
the eight-treasure pill can inhibit the occurrence and the development of liver cancer: as seen by obtaining liver tissue specimens through gross dissection, the size and the number of the liver tumors of the rats in the liver cancer model administration group are obviously smaller than those in the model group, which indicates that the eight-treasure pill can inhibit the occurrence and the development of the liver tumors of the rats (figures 1A and 1B).
The eight-treasure pill can protect liver from being damaged: the liver function index detected by serum shows that the Babaodan can obviously reduce the numerical values of alanine Aminotransferase (ALT) and aspartate Aminotransferase (AST), and the Babaodan is proved to be capable of effectively protecting the liver from being damaged (figure 1C); the result of HE staining of the pathological liver section shows that the eight-treasure pill can significantly reduce the liver inflammatory response (fig. 1D).
Example 2
Effect of Babaodan on tumorigenesis of LPS-treated Hepatic Precursor Cells (HPC) (LPS-treated hepatic precursor cells are abbreviated as LPS-HPC).
The preparation method of the LPS-HPC physiological saline cell suspension comprises the following steps: rat liver precursor cell line WB-F344 was placed in a culture medium of Dulbecco's modified Eagle's medium (DEME) containing 10% fetal bovine serum and was subjected to 5% CO conversion at 37 ℃ in the presence of 10% fetal bovine serum 2 Culturing in incubator, changing culture medium once for 2-3 days, allowing cells to grow adherent to single layer, digesting with 0.5% trypsin, subculturing with experiment group cells containing Lipopolysaccharide (LPS) with concentration of 10 μ g/mL and DEME culture medium containing 10% fetal calf serum, culturing for 2 weeks, digesting with 0.5% trypsin, and introducing to obtain culture medium containing serum extractSaline water according to 5X 10 6 Individual cells/mL concentration were resuspended for subsequent experiments. After 60 SD rats were acclimatized for one week, they were randomly divided into 6 groups, i.e., a Normal control group (Normal ctrl group, 10), a diethylnitrosamine group (DEN group, 10), a postoperative recurrence model group (DEN + LPS-HPC group, 10), a pre-BBD group (pre-BBD group, 10), a post-BBD group (post-BBD group, 10), and a full-BBD group (full-BBD group, 10).
The treatment of each group is as follows:
(1) Normal control group (Normal ctrl group): feeding purified water and injecting 0.2mL of normal saline into spleen at 4 weeks;
(2) Diethylnitrosamine group (DEN group): feeding purified water containing diethylnitrosamine (wherein DEN volume concentration is 1 10000) and intrasplenically injecting 0.2mL of physiological saline at 4 weeks;
(3) Postoperative recurrence model group (DEN + LPS-HPC group): purified water containing diethylnitrosamine (in which DEN was 1-10000 in volume) was fed, and 0.2mL of LPS-HPC physiological saline cell suspension (LPS-HPC-containing 1X 10 6 One);
(4) Eight-treasure pill administration front group (BBD front group): feeding purified water containing diethylnitrosamine (wherein DEN volume concentration is 1 10000), simultaneously perfusing purified water containing Babaodan (perfusing with 0.5g/kg dose of Babaodan twice a week for 4 weeks), and intrasplenically injecting 0.2mL LPS-HPC physiological saline cell suspension (containing LPS-HPC 1 × 10) at 4 weeks 6 And (2)) stopping the baobandan gavage after cell inoculation, and continuing feeding purified water containing diethylnitrosamine (wherein the volume concentration of DEN is 1:10000 To the end of the experiment by week 10;
(5) Eight-treasure pill administration group (BBD group): purified water containing diethylnitrosamine (in which the DEN volume concentration was 1 10000) was fed and 0.2mL of LPS-HPC physiological saline cell suspension (LPS-HPC-containing 1X 10) was intraperitoneally injected at week 4 6 And 4), meanwhile, after 4 weeks of inoculation of LPS-HPC, the stomach is filled with purified water containing Babaodan (0.5 g/kg dose of Babaodan is filled with the stomach twice a week for 6 weeks), and meanwhile, purified water containing diethylnitrosamine (wherein the DEN volume concentration is 1:10000 To the end of the experiment by week 10;
(6) The eight-treasure pill administration whole course group (BBD whole course group): during the experiment, pure water containing diethylnitrosamine (DEN volume concentration is 1 6 One).
In the experiment, a liver is injured by DEN for 4 weeks, and then an HPC simulation liver cancer recurrence model is inoculated to observe the influence of the eight-treasure pill on the formation of the liver cancer. The time for inoculating the liver cancer precursor cells is taken as a division point, and the characteristic conditions of the eight-treasure pill on the liver cancer diseases of clinical postoperative relapse patient groups are truly simulated. After 10 weeks from the start of the experimental treatment, the liver was sampled under anesthesia for appearance observation, and the degree of hepatocyte damage was observed in a pathological section.
The experimental results are as follows:
babaodan inhibited the tumorigenic capacity of LPS-treated precursor cells (HPCs): the experimental animal roughly dissects and preliminarily observes that each administration group of the eight-treasure pill has obvious cancer inhibition effect. The effect is best when eight-treasure pill is given in the whole course. Administration before HPC only (pre-BBD group), and administration after HPC (post-BBD group) had a slightly inferior effect than full-course administration, see pre-BBD, post-BBD, and full-course BBD groups in fig. 2. The results of fig. 2 show that: the eight-treasure pill medicine can obviously reduce the number and the volume of liver tissue tumors, and shows that the eight-treasure pill medicine has obvious inhibition effect on liver cancer recurrence.
The eight-treasure pill relieves the inflammatory reaction of the liver and relieves the damage of liver cells: the HE staining of the cancer tissue shows that the infiltration and damage degree of inflammatory cells around the cancer tissue can be obviously reduced by using the eight-treasure pill in the whole process. The whole course of the observation of the HE staining of the tissues beside the cancer can obviously inhibit the inflammatory reaction of the tissues beside the cancer and relieve the damage of liver cells by using the Babaodan (figure 3).
The experimental results show that the Babaodan can obviously reduce the inflammatory cell infiltration of cancer tissues and tissues beside the cancer, reduce the damage degree of liver cells and inhibit the development of liver cancer.
The above description is only an embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be made by those skilled in the art without inventive work within the technical scope of the present invention disclosed herein are intended to be covered by the scope of the present invention.
Claims (2)
1. Use of BABAODAN in preparing medicine for preventing postoperative recurrence of hepatocarcinoma is provided.
2. The use of claim 1, wherein the eight-treasure pill comprises calculus bovis, fel Serpentis, cornu Saigae Tataricae, margarita powder, notoginseng radix, and Moschus as main ingredients.
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CN202210686172.3A CN115192617A (en) | 2016-11-28 | 2016-11-28 | Application of Babaodan in preparation of medicine for preventing relapse of early liver cancer after operation |
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CN202210686172.3A CN115192617A (en) | 2016-11-28 | 2016-11-28 | Application of Babaodan in preparation of medicine for preventing relapse of early liver cancer after operation |
CN201611065626.6A CN108114010A (en) | 2016-11-28 | 2016-11-28 | Purposes of the pill of Eight Treasures in the drug for preparing prevention early liver cancer postoperative recurrence |
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