Application of schisandrol A in preparing medicine for treating cancer embolus

Abstract

The present invention discloses the application of schisandrol A in the preparation of a medicament for treating cancer thrombus, and the cancer thrombus refers to hepatocellular carcinoma portal vein tumor thrombus. The application of schisandrol A in the preparation of a medicament for treating cancer thrombus provided by the present invention is such that schisandrol A can significantly reduce the cancer thrombus formed by the liver metastasis of PVTT cells. Schisandrol A can inhibit the metastasis of hepatocellular carcinoma cells by reducing the expression of immune factors that promote cancer cell metastasis in the microenvironment, thereby inhibiting the formation of portal vein tumor thrombus. Schisandrol A has a significant inhibitory effect on the portal vein tumor thrombus caused by the metastasis of hepatocellular carcinoma cells. Schisandrol A exerts its anti-portal vein tumor thrombus effect by reducing the blood viscosity of cancer thrombus patients, that is, by promoting blood circulation.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical preparation in medicine, and specifically relates to the application of schisandrol in the preparation of a medicament for treating cancer embolism. Background Art

[0002] Hepatocellular carcinoma (referred to as liver cancer) ranks sixth in the global incidence of malignant tumors. In China, the incidence and fatality rate of liver cancer rank fifth and second respectively among malignant tumors. Due to the lack of obvious clinical symptoms in the early stage of liver cancer, about 70% - 80% of liver cancer patients in China are in the middle and late stages at the initial stage, and the overall prognosis is not ideal. Hepatocellular carcinoma cells are prone to invade the vascular system in the liver and often colonize in the portal vein and its branches. Approximately 44% - 62.2% of liver cancer patients develop into portal vein tumor thrombus (PVTT). Once PVTT appears, liver cancer patients will develop intrahepatic and extrahepatic metastases, portal hypertension, jaundice, and ascites within a short period of time, and the average median survival period is only 2.7 months. Therefore, PVTT is an important factor leading to poor prognosis of liver cancer and brings a heavy economic burden to society.

[0003] The evidence-based medical evidence for the diagnosis and treatment of liver cancer complicated with PVTT is gradually increasing. As the Chinese classification standard for PVTT, the Cheng's classification is divided into types I - IV according to the range of PVTT invading the portal vein system. Type I, the cancer thrombus invades the portal vein branches of the hepatic lobe or hepatic segment; Type II, the cancer thrombus invades the left or right branch of the portal vein; Type III, the cancer thrombus invades the main trunk of the portal vein; Type IV, the cancer thrombus invades the superior mesenteric vein; Type I0, microvascular cancer thrombus is diagnosed by postoperative pathology. There is no consensus on the rational selection of treatment methods according to different types of portal vein cancer thrombi. Since it is difficult to remove portal vein cancer thrombi by surgery, corresponding drug treatments to inhibit the metastasis of liver cancer and the formation of portal vein cancer thrombi are very necessary. Currently, for patients with unresectable liver cancer complicated with PVTT, in addition to the commonly used chemotherapeutic drugs for inhibiting the metastasis of liver cancer, such as fluorouracil and oxaliplatin, and the molecular targeted drugs sorafenib and lenvatinib. Regorafenib is a second-line treatment drug applicable to patients who have failed sorafenib or lenvatinib treatment. Large-scale, multi-center clinical controlled experiments have shown that the incidence rate of PVTT with a hypercoagulable state in liver cancer patients is higher and the range is wider, which seriously affects the survival expectancy of cancer patients. Literature reports that the antiplatelet drug aspirin can reduce the incidence of liver cancer after surgery in patients with viral hepatitis-related liver cirrhosis and improve the long-term survival rate. Therefore, finding drugs to reduce the formation of cancer thrombi and inhibit the metastasis of liver cancer in the portal vein system is of great significance for improving the survival rate and quality of life of patients.

[0004] Plants of the genus Schisandra in Magnoliaceae are one of the important groups of medicinal plant resources in China. Schisanhenol (SAL) is a dibenzocyclooctene lignan with relatively high content in plants of this genus. It can be extracted from the dried mature fruits of Schisandra rubriflora Rehd. et Wils., Schisandra chinensis (Turcz.) Baill. (also known as Schisandra chinensis in the north), and Schisandra sphenanthera Rehd. et Wils., and has the effects of astringing and arresting sweating, replenishing qi and promoting fluid production, tonifying the kidney and tranquillizing the mind. The fruits of Schisandra show antagonistic effects on platelet aggregation induced by arachidonic acid, collagen, platelet-activating factor, and thrombin. Its lignans and their derivatives are antagonists of platelet-activating factor. There have been literature reports that schisanhenol can inhibit adenosine diphosphate- and platelet-activating factor-induced rabbit platelet aggregation in vitro. Patents on schisanhenol at home and abroad mostly focus on the treatment of Parkinson's disease, ischemic stroke, antidepressant effects, prevention and treatment of diarrhea caused by mycophenolic acid, etc. The patent application with the publication number CN1723879A, titled "Use of Schisanhenol in the Preparation of Drugs for Treating Ischemic Stroke", discloses that schisanhenol isolated from the fruits of Schisandra rubriflora produced in Lijiang, Yunnan, has anticoagulant and fibrinolytic, antiplatelet aggregation, and vasodilating activities in vitro, and has a therapeutic and protective effect on cerebral infarction and reperfusion injury in terms of brain function and morphology. The patent application with the publication number CN104546705A, titled "A New Use of Schisanhenol as a Drug and a Preparation Method of Its Homogeneous Suspension", discloses that schisanhenol is formulated into a homogeneous suspension. Through animal experiments, it is found that in an animal model of Parkinson's disease induced by MPTP, the homogeneous suspension of schisanhenol can significantly improve the motor disorders of experimental animals and shows a good dose-dependent trend.

[0005] The patent application with the publication number CN102627625A, titled "Derivatives of Schisandrol A, Schisandrin B and Schisandrin C and Their Uses", discloses the structural modification and transformation of schisandrin B, etc., with halogenation and / or oxidation carried out at the 4-substituted position and / or 11-substituted position respectively. The resulting derivatives have a significant protective effect on CCl4-induced liver injury in human liver HL-7702 cells and can be used as candidate compounds for novel anti-liver injury drugs, especially anti-chemical injury drugs. The patent application with the publication number CN108472502A, titled "Compositions, Methods and Pharmaceutical Compositions for Treating and Maintaining Liver Health", discloses a composition for treating and maintaining liver health, which includes a mixture of plant extracts, wherein the plant extracts contain at least one extract of Myristica, at least one extract of Astragalus, and at least one extract of Poria or at least one extract of Schizandra. However, it does not involve the evaluation of the anti-cancer embolism formation and metastasis effects of schisandrin B on the whole animal model in terms of liver portal vein cancer embolism.

[0006] Looking at the research reports at home and abroad, there is no literature report on the effect of schisandrin B in treating liver portal vein cancer embolism. Summary of the Invention

[0007] The purpose of the present invention is to provide an application of schisandrin B in the preparation of a drug for treating cancer embolism.

[0008] In order to achieve the above purpose, the technical scheme adopted by the present invention is as follows:

[0009] The first aspect of the present invention provides an application of schisandrin B in the preparation of a drug for treating cancer embolism.

[0010] The cancer embolism refers to liver cancer portal vein cancer embolism.

[0011] The drug for treating cancer embolism uses schisandrin B as the only active ingredient.

[0012] The schisandrin B can be made into a pharmaceutical preparation.

[0013] The dosage form of the pharmaceutical preparation is selected from at least one of powder, tablet, granule, capsule, solution, emulsion, and suspension.

[0014] The administration method of the drug is oral administration.

[0015] The structural formula of the schisandrin B is as follows:

[0016]

[0017] The application of schisandrol A provided by the present invention in the preparation of a medicament for treating cancer embolism. The present invention is based on an animal model in which a cell line PVTT of human portal vein cancer thrombus tissue induces the formation of liver cancer portal vein cancer thrombus in BALB / c-nu nude mice. Through experiments on the animal model of liver cancer portal vein cancer thrombus, it is confirmed that schisandrol A has the pharmacological effect of anti-liver cancer portal vein cancer thrombus and can be used to develop a medicament for treating liver cancer complicated with portal vein cancer thrombus. Through the mouse tail-cutting experiment, it is proved that schisandrol A has the effect of promoting blood circulation.

[0018] Due to the adoption of the above technical solution, the present invention has the following advantages and beneficial effects:

[0019] The application of schisandrol A provided by the present invention in the preparation of a medicament for treating cancer embolism. Schisandrol A can significantly reduce the cancer thrombus formed by the liver metastasis of PVTT cells. Schisandrol A can inhibit the metastasis of liver cancer cells by reducing the expression of immune factors that promote cancer cell metastasis in the microenvironment, thereby inhibiting the formation of portal vein cancer thrombus; Schisandrol A has a significant inhibitory effect on the portal vein cancer thrombus caused by the metastasis of liver cancer cells. Schisandrol A exerts the effect of anti-portal vein cancer thrombus by reducing the blood viscosity of cancer thrombus patients, that is, the effect of promoting blood circulation. Description of the Drawings

[0020] Figure 1 It is a schematic diagram of the HE staining result of the mouse liver portal vein tissue section of the present invention.

[0021] Figure 2 It is a schematic diagram of the percentage of the metastatic cancer thrombus area calculated by the ImageProPlus software of the present invention.

[0022] Figure 3 It is a schematic diagram of the measurement result of the TGF-β1 content in the serum of the present invention.

[0023] Figure 4 It is a schematic diagram of the Western Blot evaluation result of the E-CAD protein in the liver cancer tissue of the present invention.

[0024] Figure 5 It is a schematic diagram of the statistical result of the bleeding volume of mice in different groups of the present invention through the tail-cutting experiment. Detailed Embodiments

[0025] In order to more clearly illustrate the present invention, the present invention will be further described below in conjunction with preferred embodiments. Those skilled in the art should understand that the specific content described below is illustrative rather than restrictive, and should not be used to limit the protection scope of the present invention.

[0026] Example 1

[0027] Establishment of an animal model for inducing hepatocellular carcinoma portal vein tumor thrombus in BALB / c-nu nude mice by the cell line PVTT (tumor thrombus type hepatocellular carcinoma cells) cultured from the portal vein tumor thrombus tissue of liver cancer patients, including the following steps:

[0028] Culture of PVTT cell line and Control Liver Cancer (CLC) cell line: The Shanghai Eastern Hepatobiliary Surgery Hospital (the Third Affiliated Hospital of Naval Medical University) provided the cell line PVTT (tumor thrombus type hepatocellular carcinoma cells) cultured from the portal vein tumor thrombus tissue of liver cancer patients and the cell line CLC (non-tumor thrombus type hepatocellular carcinoma cells) cultured from the in-situ hepatocellular carcinoma tissue.

[0029] Cell culture: The cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin double antibody in an incubator at 37 °C and 5% CO 2 .

[0030] Cell passage: When the cell density reached 90%, the original culture medium was discarded, the cells were washed once with PBS, the PBS was discarded, 2 mL of 0.25% trypsin was added to the culture dish, digested for 3 min, 2 mL of complete medium was added to terminate the digestion, the cells were gently pipetted and mixed with a 5 mL pipettor, transferred to a 15 mL centrifuge tube, centrifuged at 1300 rpm for 3 min, the supernatant was discarded, and the cells were resuspended with complete medium in the centrifuge tube and passaged at a ratio of 1:2.

[0031] Establishment of the portal vein tumor thrombus animal model and hepatocellular carcinoma animal model: Five-week-old male BALB / c-nu nude mice were randomly divided into 3 groups of 8 mice each. The cell line PVTT (tumor thrombus type hepatocellular carcinoma cells) cultured from the portal vein tumor thrombus tissue of liver cancer patients with good growth status was digested and collected with 0.25% trypsin, washed with PBS, counted, and resuspended to a concentration of 1.67×10 8 cells / mL and kept on ice to maintain cell viability. The mice were anesthetized with 1.0% isoflurane gas, the side abdomen of the mice was cut open with ophthalmic scissors, the spleen of the mice was pulled out with ophthalmic forceps, 30 μL of cell suspension was aspirated with an insulin syringe and injected into the spleen of the mice, then the spleen was put back in place, and the wound was sutured with 4-0 suture. Drugs were administered 24 h after modeling to establish the portal vein tumor thrombus animal model. The same operation was performed with the cell line CLC cultured from the in-situ hepatocellular carcinoma tissue to obtain the hepatocellular carcinoma animal model.

[0032] Example 2

[0033] Schisanhenol used in the present invention was purchased from Shanghai Yuanye Bio-Technology Co., Ltd.

[0034] The experiment on schisanhenol against hepatocellular carcinoma portal vein tumor thrombus includes the following steps:

[0035] (1) Dosage of drug administration:

[0036] They were divided into three groups: the non-cancer thrombus type hepatocarcinoma cell CLC group (the control group, i.e., the CLC group), the cancer thrombus type hepatocarcinoma cell group (the model group, i.e., the PVTT group), and the cancer thrombus type hepatocarcinoma cell schizandrol treatment group (the administration group, i.e., the Schi. group).

[0037] On the basis of the model obtained in Example 1, the non-cancer thrombus type hepatocarcinoma cell CLC group (the control group, i.e., the CLC group) and the cancer thrombus type hepatocarcinoma cell group (the model group, i.e., the PVTT group) were intragastrically administered 0.5% CMC-Na (carboxymethyl cellulose sodium) every two days. The cancer thrombus type hepatocarcinoma cell schizandrol treatment group (the administration group, i.e., the Schi. group) was intragastrically administered 1 mg / kg of schizandrol every two days for six consecutive weeks.

[0038] (2) Tissue sample collection: Four mice were randomly selected from each group for HE staining of liver tissue sections to observe the metastasis of portal vein cancer thrombus cells in blood vessels; reduce the number of cancer thrombi in veins. After sacrificing the mice by cervical dislocation at the sixth week, the abdominal cavity was opened, and the whole liver was removed and immersed in 4% paraformaldehyde for 48 hours for fixation.

[0039] (3) HE staining tissue fixation and sectioning: First, the mouse liver tissue samples were longitudinally placed and embedded in paraffin to make it easier to obtain longitudinal sections of large blood vessels. Liquid paraffin was added to the mold, and the tissue samples to be embedded were placed in the paraffin to ensure regular tissue position. After adding a little liquid paraffin, it was cooled and frozen to make the paraffin become solid to achieve the effect of tissue fixation, and then a paraffin slicer was used for sectioning, with a thickness of about 4 - 8 μm. The cut tissue sections were placed on glass slides and soaked in warm water at 40°C to fully stretch the tissue.

[0040] (4) Dewaxing of HE staining samples: The tissue sample sections to be tested were placed in xylene and soaked thoroughly for 10 min. After soaking, the xylene was replaced and soaked for another 10 min. The purpose was to dissolve the paraffin in the sections with xylene, so that when staining with the dye solution, the dye solution could fully enter the tissue. At the same time, xylene could also play a role in making the sections transparent, making it easier to observe the sections.

[0041] (5) Hydration of HE staining samples: The tissue samples to be tested soaked in xylene were first placed in absolute ethanol and soaked for 5 min to wash out the xylene used for dewaxing and allow water to enter the tissue; then they were successively placed in 95%, 85%, and 70% ethanol and soaked for 5 min each to achieve the effect of full hydration.

[0042] (6)HE staining section hematoxylin staining, differentiation and blueing: Immerse the section of the hydrated tissue sample in PBS solution for cleaning, soak for 5 min each time, and clean a total of 3 times. Then use a pipette to aspirate the pre-prepared hematoxylin staining solution, add 100 ul to each tissue section, and stain fully for 10 min. After staining, wash away the excess hematoxylin staining solution with distilled water. Then use 1% hydrochloric acid ethanol for differentiation to remove the excessive dye bound in the cell nucleus and the excess dye in the cytoplasm. After differentiation, rinse the tissue section with double-distilled water. To make the hematoxylin turn blue, add a weakly alkaline blueing solution to the tissue section to stain the cell nucleus blue. After blueing, first wash with clear water, and then rinse the tissue section with double-distilled water.

[0043] (7)HE staining section eosin staining and dehydration: Add eosin staining solution to the tissue sample section of the previous step, and allow the tissue to stain fully for 3 min. After staining, dehydrate the tissue section step by step, using ethanol with concentrations of 80%, 95% and absolute ethanol respectively. Dehydrate with 80% ethanol for 5 s, 95% ethanol for 2 min, and absolute ethanol for 2 min.

[0044] (8)Air-dry and mount the HE staining sample section: Immerse the dehydrated tissue sample section in xylene twice, each time for 4 min, then dry the tissue sample section and mount it with neutral balsam.

[0045] (9)Finally, observe under the microscope. The long strip-shaped hollow part in the section is the large blood vessel. The dark purple, irregularly shaped cells inside and around the large blood vessel are the cancer cells invading the large blood vessels of the liver. Randomly select three fields of view on each section and take pictures at 5 times and 20 times respectively under the microscope. Use ImageProPlus software to calculate the percentage of the area of the metastatic cancer thrombus, that is, the ratio of the area of the cancer thrombus in each selected field of view to the area of the liver tissue in the whole field of view.

[0046] According to the above method steps, evaluate the inhibitory effect of schisandrol on the formation of cancer thrombus in the portal vein of liver cancer. The results are as Figure 1 and Figure 2 shown. Figure 1 This is a schematic diagram of the HE staining result of the mouse liver portal vein tissue section of the present invention. Figure 2 This is a schematic diagram of calculating the percentage of the area of the metastatic cancer thrombus by ImageProPlus software of the present invention. Figure 1 of the liver tissue HE staining and Figure 2From the statistical results of the cancer thrombus area, it can be concluded that compared with the cancer thrombus type liver cancer cell group (the model group, i.e., the PVTT group), the cancer thrombus type liver cancer cell schizandrol treatment group (the administration group, i.e., the Schi. group) can significantly reduce the cancer thrombus formed by the liver metastasis of PVTT cells, and there is a significant difference (P<0.01), indicating that schizandrol has a definite therapeutic effect on inhibiting the metastasis of PVTT cells to the portal vein and forming portal vein cancer thrombus.

[0047] Example 3

[0048] Determination of the content of TGF-β1 in serum includes the following steps:

[0049] (1) Making a standard curve: Prepare a standard solution with a maximum concentration of 2000 pg / mL and perform a 2-fold serial dilution to obtain a total of 8 points to make a standard curve.

[0050] (2) Soaking the enzyme-linked immunosorbent assay (ELISA) plate: Add 300 μl of 1× washing solution and soak for 30 seconds. After discarding the washing solution, pat dry on absorbent paper.

[0051] (3) Adding standards: Add 100 μl of the 2-fold serially diluted standards to the standard wells. Add 100 μl of 1× detection buffer to the blank well.

[0052] (4) Adding samples: Add 50 μl of 1× detection buffer and 50 μl of the sample to the sample wells.

[0053] (5) Adding detection antibody: Add 50 μl of the diluted detection antibody (diluted 1:100) to each well

[0054] (6) Incubation: Seal the plate with a sealing film, shake at 300 revolutions per minute, and incubate at room temperature for 1.5 hours.

[0055] (7) Washing: Discard the liquid, add 300 μl of washing solution to each well to wash the plate, and wash 6 times.

[0056] (8) Adding enzyme and incubating: Add 300 μl of washing solution to each well to wash the plate, wash 6 times, and pat dry on absorbent paper each time after washing the plate.

[0057] (9) Incubation: Seal the plate with a new sealing film. Shake at 300 revolutions per minute and incubate at room temperature for 30 minutes.

[0058] (10) Repeat step 6.

[0059] (11) Adding substrate for color development: Add 100 μl of the chromogenic substrate TMB (3,3',5,5'-tetramethylbenzidine) to each well, protect from light, and incubate at room temperature for 8 minutes.

[0060] (12) Adding stop solution: Add 100 μl of stop solution to each well, and the color changes from blue to yellow.

[0061] (13) Detection reading: Use an enzyme-linked immunosorbent assay (ELISA) reader to perform double-wavelength detection within 30 minutes, and measure the OD values at the maximum absorption wavelength of 450 nm and the reference wavelength of 570 nm. Subtract the measured value at 570 nm from the measured value at 450 nm to obtain the calibrated OD value.

[0062] (14) Result calculation: Calculate the average OD values of the standard products and samples, then subtract the OD value of the zero-concentration standard product, and calculate the measured concentration of TGF-β1 according to the standard curve.

[0063] The ELISA results of mouse serum are as Figure 3 shown, Figure 3 which is a schematic diagram of the measurement results of the TGF-β1 content in the serum of the present invention. It can be Figure 3 concluded that the content of TGF-β1 in the serum of mice in the cancer thrombus type hepatocarcinoma cell group (model group, i.e., PVTT group) is significantly increased compared with that in the non-cancer thrombus type hepatocarcinoma cell CLC group (control group, i.e., CLC group), while the content of TGF-β1 in the serum of the cancer thrombus type hepatocarcinoma cell schisandrol treatment group (drug administration group, i.e., Schi. group) is significantly decreased compared with that in the cancer thrombus type hepatocarcinoma cell group (model group, i.e., PVTT group). This shows that schisandrol can inhibit the metastasis of hepatocarcinoma cells by reducing the expression of immune factors that promote cancer cell metastasis in the microenvironment, and thus inhibit the formation of portal vein cancer thrombus.

[0064] Example 4

[0065] Western Blot evaluation of tissue E-CAD (E-cadherin) protein includes the following steps:

[0066] (1) Protein sample preparation: Cut 10 mg of tissue, add 550 μl of RIPA lysis buffer containing protease inhibitor, grind it into a homogenate, lyse it at 4°C for 15 minutes, then centrifuge at 14000 g for 15 minutes. Carefully transfer the supernatant to a new 1.5 mL EP tube, add 5× protein electrophoresis buffer and mix well, then boil for 5 minutes to denature the protein.

[0067] (2) Protein concentration determination: Before adding 5× protein electrophoresis buffer and boiling for denaturation, use a BCA protein quantification kit to measure the concentration of each sample and calculate the loading amount.

[0068] (3) Electrophoresis: Load 25 μg of each sample, and add protein molecular weight marker on both sides of the sample wells. First, perform constant voltage electrophoresis at 90 V until the sample runs out of the stacking gel, then adjust the voltage to 120 V, and stop electrophoresis when the bromophenol blue runs to the lower edge of the gel.

[0069] (4) Wet transfer of membrane: Cut a PVDF membrane that matches the size of the gel and soak it in methanol for 2 minutes for activation. Place the filter paper, sponge, and the electrophoresed gel in the electrotransfer buffer for wetting and equilibration. Make a "sandwich" in the order of sponge - filter paper - PVDF membrane - gel - filter paper - sponge, and place it in the electrotransfer tank in the correct direction (the gel side faces the black electrotransfer tank at the negative electrode, and the PVDF membrane side faces the red electrotransfer tank at the positive electrode). Fill the electrotransfer tank with the electrotransfer buffer, place the electrotransfer tank device in an ice bath environment, and perform electrotransfer at a constant current of 230 mA for about 60 minutes.

[0070] (5) Blocking: After the electrotransfer is completed, wash the PVDF membrane with TBST and block it with a rapid blocking solution for 15 min.

[0071] (6) Immune reaction: Immerse the blocked PVDF membrane completely in the primary antibody working solution and incubate it overnight on a shaker at 4°C. Recover the primary antibody, wash the membrane 3 times with TBST for 5 minutes each time, add the fluorescent secondary antibody working solution, and incubate it in the dark at room temperature for 1 hour. Recover the secondary antibody, wash the membrane 3 times with TBST for 5 minutes each time.

[0072] (7) Detection: Place the PVDF membrane in an Odyssey dual-color infrared fluorescence imaging system for membrane scanning.

[0073] Figure 4 This is a schematic diagram of the Western Blot evaluation results of E-CAD protein in liver cancer tissues of the present invention. The loss of E-CAD is one of the key features of the EMT (epithelial-mesenchymal transition) process, and EMT is considered a pre-metastatic cellular event that promotes tumor cell invasion and malignant tumor progression. Figure 4 Protein quantification of WB in mouse liver tissues shows that the expression level of E-CAD in the cancer thrombus type liver cancer cell group (the model group, i.e., the PVTT group) is significantly lower than that in the non-cancer thrombus type liver cancer cell CLC group (the control group, i.e., the CLC group), and the expression level of E-CAD in the cancer thrombus type liver cancer cell schizandrol treatment group (the drug administration group, i.e., the Schi. group) significantly rebounds, indicating that schizandrol has a significant inhibitory effect on portal vein cancer thrombus caused by liver cancer cell metastasis. Figure 4 In [description], GAPDH is an enzyme in the glycolysis reaction, which is widely distributed in cells of various tissues and serves as an internal reference protein in this experiment.

[0074] Example 5

[0075] The schizandrol mouse tail blood circulation promotion experiment includes the following steps:

[0076] (1) Animal administration: Male C57BL / 6 mice at 7 weeks of age were selected and randomly divided into 6 groups with 8 mice in each group. Each mouse in the blank group was given 0.5% CMC-Na solution, and each mouse in the schisandrol group was given schisandrol at 3 mg / kg / day, 10 mg / kg / day, 30 mg / kg / day, and 60 mg / kg / day respectively. Each mouse in the Warfarin group was given warfarin at 5 mg / kg / day. The administration was continued for 4 days.

[0077] (2) Tail cutting experiment: 4 hours after the last administration, the mice were anesthetized and placed on a thermostatic operating table. Their limbs were clamped, and the tails of the mice were cut off with scissors at a distance of 1.5 cm from the tip and quickly placed into 10 ml of physiological saline at a constant temperature of 37°C. After soaking for 10 minutes, the mouse tails were quickly taken out.

[0078] (3) Red blood cell treatment: 5 ml of red blood cell lysis buffer salt (containing 155 mM NH 4 Cl, 10 mM KHCO 3 , 0.1 mM Na 2 EDTA, pH adjusted to 7.3) was added to the above solution to lyse the red blood cells in the blood and shaken evenly.

[0079] (4) Establishment of the blood volume standard curve for mice: 0, 10, 30, 100, 300, and 600 μl of fresh mouse blood were respectively added to 10 ml of physiological saline at 37°C. 5 ml of red blood cell lysis buffer salt was added to lyse the red blood cells in the blood and shaken evenly. After placing in a 96-well plate, the absorbance value was measured with an enzyme-linked immunosorbent assay (ELISA) reader at 405 nm to establish a standard curve.

[0080] (5) Detection and calculation of blood volume: 50 μl of samples from different groups were taken out respectively and placed in a 96-well plate. The absorbance value was measured with an ELISA reader at 405 nm, and the blood volume (μl) was calculated using the standard curve.

[0081] According to the above method steps, the blood-activating effect of schisandrol on mice was evaluated. The results are as Figure 5 shown, Figure 5 This is a schematic diagram of the statistical results of the blood loss of mice in different groups of this invention after the tail cutting experiment. Figure 5It can be concluded that compared with the blank group, schisandrol A (30 mg / kg / day and 60 mg / kg / day) can significantly increase the blood loss of mice in a dose-dependent manner (P<0.05); the blood loss caused by the schisandrol A 60 mg / kg / day group is better than that of the warfarin 5 mg / kg / day group. This shows that both schisandrol A and warfarin have a certain effect of reducing blood viscosity after oral administration to mice and are potential blood-activating drugs. Schisandrol A may play a role in anti-portal vein cancer thrombus by reducing the blood viscosity of cancer thrombus patients, that is, by activating blood circulation.

[0082] The above are only the preferred embodiments of the present invention, and do not impose any form of limitation on the present invention. Although the present invention has been disclosed above with the preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art of this patent can make some changes or modifications to equivalent embodiments by using the technical content prompted above within the scope of the technical solution of the present invention. However, as long as it does not deviate from the content of the technical solution of the present invention, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention still fall within the scope of the present invention's solution.

Claims

1. Use of schisandrol A in the preparation of a medicament for treating cancer embolism, Characterized in that, The cancer embolism refers to portal vein cancer embolism of liver cancer; The medicament for treating cancer embolism uses schisandrol A as the sole active ingredient.

2. The use of schisandrol A in the preparation of a medicament for treating cancer embolism according to claim 1, Characterized in that, The dosage form of the medicament is selected from at least one of powder, tablet, granule, capsule, solution, emulsion, suspension.

3. The use of schisandrol A in the preparation of a medicament for treating cancer embolism according to claim 2, Characterized in that, The administration method of the medicament is oral administration.

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