Traditional Chinese medicine composition for treating lung cancer and application thereof

Abstract

The invention relates to a traditional Chinese medicine composition for treating lung cancer and application of the traditional Chinese medicine composition in inhibiting or reducing lung cancer cell viability, the traditional Chinese medicine composition comprises 3-30 parts by weight of dahurian angelica root, 3-30 parts by weight of baical skullcap root, 3-30 parts by weight of semen lepidii, 3-30 parts by weight of apricot kernel and 3-30 parts by weight of mirabilite. The content of the semen lepidii is 8-35 parts by weight, the content of the almond is 5-30 parts by weight, and the content of the mirabilite is 0.3-5 parts by weight, and cell experiments prove that the traditional Chinese medicine composition is applied to lung cancer cells in an effective dose of 1-100 [mu] g / ml so as to induce protein expression of LC3-I / LC3-II, so that the lung cancer cells can be effectively treated. The autophagy effect of the lung cancer cells is initiated, so that the activity of the lung cancer cells is inhibited or reduced.

Description

Technical Field

[0001] The invention relates to a traditional Chinese medicine for treating lung cancer, and in particular to a traditional Chinese medicine composition for treating lung cancer and the use of the composition in inhibiting or reducing the activity of lung cancer cells. Background Art

[0002] Lung cancer has a high mortality rate because it is difficult to diagnose in the early stages. When patients are found to have lung cancer cells, most of the lung cancer cells have already metastasized, and the treatment effect is limited. Currently, common treatments for lung cancer include: lung function resection surgery, radiotherapy, and chemotherapy. Among them, chemotherapy is to administer targeted drugs to patients to inhibit the growth of lung cancer cells, slow down the spread of lung cancer cells, and prevent cancer recurrence.

[0003] However, the targeted drugs currently used in chemotherapy can have significant side effects on the human body. In order to improve the side effects of cancer drugs, some related industries have developed natural compounds, such as Chinese herbal medicines. Many studies have pointed out that there are many specific Chinese herbal medicines that can serve as adjuvant therapy in the treatment of advanced non-small cell lung cancer (NSCLC).

[0004] For example, many research papers have shown that Scutellariae radix and Angelica dahurica have certain effects on cancer treatment. In short, Scutellariae radix can not only increase the number of immunoglobulins in patients to enhance their immunity, but the Scutellariae radix extract has been shown to be cytotoxic to non-small cell lung cancer cells and can significantly inhibit the metastasis of non-small cell lung cancer cells. Angelica dahurica contains the active ingredient furanocoumarin, which can promote the apoptosis of human lung cancer cells and has the effect of treating cancer.

[0005] However, although the above studies have confirmed that the natural components of certain Chinese herbal medicines have the effect of inhibiting lung cancer cells, the efficacy of individual Chinese herbal medicines against lung cancer is still limited. Therefore, how to select a combination of Chinese herbal medicines to effectively treat lung cancer is the purpose of the present invention. Summary of the invention

[0006] In view of this, the purpose of the present invention is to provide a traditional Chinese medicine composition for treating lung cancer and its preparation method for inhibiting or reducing the activity of lung cancer cells. The traditional Chinese medicine composition is composed of Angelica dahurica, Scutellaria baicalensis, Lemongrass seeds, Apricot kernel and Sodium sulfate, which can effectively induce autophagy of the non-small cell lung cancer and small cell lung cancer, thereby inhibiting or reducing the number of active cells of the non-small cell lung cancer and small cell lung cancer.

[0007] In order to achieve the above-mentioned object, the present invention provides a traditional Chinese medicine composition for treating lung cancer, comprising Angelica dahurica, Scutellaria baicalensis, Lemongrass seeds, Apricot kernel and Sodium sulfate, wherein the content of the traditional Chinese medicine composition is calculated by weight, the content of Angelica dahurica is between 3 and 30 parts by weight, the content of Scutellaria baicalensis is between 3 and 30 parts by weight, the content of Lemongrass seeds is between 8 and 35 parts by weight, the content of Apricot kernel is between 5 and 30 parts by weight, and the content of Sodium sulfate is between 0.3 and 5 parts by weight.

[0008] In one embodiment, the total content of the Angelica dahurica and the Scutellaria baicalensis is a first component amount, the total content of the Lemongrass seeds, the Apricot kernel and the Sodium Sulfate is a second component amount, and the first component amount is greater than the second component amount.

[0009] In one embodiment, the content of Angelica dahurica is between 5 and 10 parts by weight, the content of Scutellaria baicalensis is between 10 and 15 parts by weight, the content of Lemongrass seeds is between 10 and 15 parts by weight, the content of Apricot kernel is between 8 and 12 parts by weight, and the content of Sodium sulfate is between 0.5 and 1.5 parts by weight.

[0010] In one embodiment, the content of Angelica dahurica is between 8 and 10 parts by weight, the content of Scutellaria baicalensis is between 13 and 15 parts by weight, the content of Lemongrass seeds is between 10 and 12 parts by weight, the content of Apricot kernel is between 8 and 10 parts by weight, and the content of Sodium sulfate is between 1 and 1.5 parts by weight.

[0011] In one embodiment, the total content of the scutellaria baicalensis and the scutellaria baicalensis seeds is a third component amount, the total content of the angelica dahurica, the apricot kernel and the mirabilite is a fourth component amount, and the third component amount is greater than the fourth component amount.

[0012] In one embodiment, the content of Angelica dahurica is between 5 and 10 parts by weight, the content of Scutellaria baicalensis is between 20 and 30 parts by weight, the content of Lemongrass seeds is between 20 and 30 parts by weight, the content of Apricot kernel is between 8 and 12 parts by weight, and the content of Sodium sulfate is between 0.5 and 1.5 parts by weight.

[0013] In one embodiment, the total content of the Angelica dahurica and the Scutellaria baicalensis is a first component amount, the total content of the Lemongrass seeds, the Apricot kernel and the Sodium Sulfate is a second component amount, and the second component amount is greater than the first component amount.

[0014] In one embodiment, the content of Angelica dahurica is between 15 and 25 parts by weight, the content of Scutellaria baicalensis is between 10 and 15 parts by weight, the content of Lemongrass seeds is between 20 and 30 parts by weight, the content of Apricot kernel is between 15 and 25 parts by weight, and the content of Sodium sulfate is between 2 and 4 parts by weight.

[0015] The present invention provides another embodiment of the use of the aforementioned Chinese medicine composition for preparing a method for inhibiting or reducing the activity of lung cancer cells, wherein the Chinese medicine composition is administered to the lung cancer cells at an effective dose of 1 to 100 μg / ml to induce the protein expression of LC3-I / LC3-II and trigger the autophagy of the lung cancer cells.

[0016] In one embodiment, the effective dose of the Chinese medicine composition is between 25 and 100 μg / ml, and the lung cancer cells include non-small cell lung cancer cells or small cell lung cancer cells.

[0017] The effect of the present invention is that the Chinese medicine composition is composed of specific weight portions of Angelica dahurica, Scutellaria baicalensis, Lemongrass seeds, Apricot kernel and Sodium sulfate and a Chinese medicine compound. Cell experiments have confirmed that the pharmacological addition and subtraction effects of Angelica dahurica, Scutellaria baicalensis, Lemongrass seeds, Apricot kernel and Sodium sulfate in the Chinese medicine composition can indeed more effectively inhibit and reduce the number of active non-small cell lung cancer cells and small cell lung cancer cells in a short period of time, and the Chinese medicine composition can significantly induce the expression of LC3-I / LC3-II proteins in non-small cell lung cancer cells and small cell lung cancer cells under the condition of an effective dose of 25 to 100 μg / ml, thereby inducing the autophagy of the lung cancer cells, thereby achieving the desired therapeutic effect of treating or improving lung cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1A The figure is a data graph showing the cell survival rate of lung adenocarcinoma cells (H460) corresponding to different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml when Scutellaria baicalensis was used alone.

[0019] Figure 1B The electrophoresis diagram of LC3-I / LC3-II proteins in lung adenocarcinoma cells (H460) corresponding to different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml of Scutellaria baicalensis alone.

[0020] Figure 2A The figure is a data graph showing the cell survival rate of lung adenocarcinoma cells (H460) corresponding to different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml of Angelica dahurica alone.

[0021] Figure 2B The electrophoresis diagram of LC3-I / LC3-II proteins in lung adenocarcinoma cells (H460) corresponding to different effective doses of 0μg / ml, 1μg / ml, 5μg / ml, 25μg / ml, 50μg / ml, and 100μg / ml of Angelica dahurica alone.

[0022] Figure 3A The figure is a data graph showing the cell survival rate of lung adenocarcinoma cells (H460) corresponding to different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml using a combination of Scutellaria baicalensis and Angelica dahurica.

[0023] Figure 3B The electrophoresis diagram of LC3-I / LC3-II proteins in lung adenocarcinoma cells (H460) corresponding to different effective doses of 0μg / ml, 1μg / ml, 5μg / ml, 25μg / ml, 50μg / ml, and 100μg / ml of the combination of Scutellaria baicalensis and Angelica dahurica.

[0024] Figure 4A The figure is a data graph showing the cell survival rate of lung adenocarcinoma cells (H460) corresponding to different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml of the Chinese medicine composition in a preferred embodiment of the present invention.

[0025] Figure 4B The figure is the electrophoresis diagram of LC3-I / LC3-II proteins of lung adenocarcinoma cells (H460) corresponding to different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml of the Chinese medicine composition in a preferred embodiment of the present invention.

[0026] Figure 5A The figure is a data graph showing the cell survival rate of small cell lung cancer cells (H1355) corresponding to different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml when Scutellaria baicalensis is used alone.

[0027] Figure 5B The LC3-I / LC3-II protein electrophoresis diagram of small cell lung cancer cells (H1355) corresponding to the administration of Scutellaria baicalensis alone at different effective doses of 0μg / ml, 1μg / ml, 5μg / ml, 25μg / ml, 50μg / ml, and 100μg / ml.

[0028] Fig. 6A The graph is a data graph of cell survival rate of small cell lung cancer cells (H1355) corresponding to different effective doses of 0μg / ml, 1μg / ml, 5μg / ml, 25μg / ml, 50μg / ml, and 100μg / ml of Angelica dahurica alone.

[0029] Figure 6BThe LC3-I / LC3-II protein electrophoresis diagram of small cell lung cancer cells (H1355) corresponding to the different effective doses of 0μg / ml, 1μg / ml, 5μg / ml, 25μg / ml, 50μg / ml, and 100μg / ml of Angelica dahurica alone.

[0030] Fig. 7A The figure is a data graph showing the cell survival rate of small cell lung cancer cells (H1355) corresponding to different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml using a combination of Scutellaria baicalensis and Angelica dahurica.

[0031] Figure 7B The LC3-I / LC3-II protein electrophoresis diagram of small cell lung cancer cells (H1355) corresponding to different effective doses of 0μg / ml, 1μg / ml, 5μg / ml, 25μg / ml, 50μg / ml, and 100μg / ml using the combination of Scutellaria baicalensis and Angelica dahurica.

[0032] Fig. 8A The figure is a data graph showing the cell survival rate of small cell lung cancer cells (H1355) corresponding to different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml of the Chinese medicine composition in a preferred embodiment of the present invention.

[0033] Figure 8B The figure is the LC3-I / LC3-II protein electrophoresis diagram of small cell lung cancer cells (H1355) corresponding to different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml of the Chinese medicine composition in a preferred embodiment of the present invention. DETAILED DESCRIPTION

[0034] A preferred embodiment of the present invention provides a traditional Chinese medicine composition for treating lung cancer, the basic ingredients of which include Angelica dahurica, Scutellaria baicalensis, Lemongrass seeds, Apricot kernel and Sodium Sulfate, wherein the Angelica dahurica, Scutellaria baicalensis, Lemongrass seeds and Apricot kernel in the traditional Chinese medicine composition are all extracted from natural plants.

[0035] The pharmacological properties of each component in the Chinese medicine composition are as follows:

[0036] Angelica dahurica: The active ingredients include angelicae dahuricae, angelicae ether and angelicae dahuricae coumarin, which have antipyretic and analgesic, antibacterial and anti-inflammatory, antioxidant and other effects, and have certain inhibitory effects on bacteria, molds and yeasts.

[0037] Scutellaria baicalensis: The active ingredients include acacetin, apigenin, baicalin, chrysin and wogonin. Each of these active ingredients has certain special therapeutic effects, such as anti-cancer, liver protection, anti-cardiovascular disease, anti-allergy and anti-inflammatory effects, and also has anti-tumor effects.

[0038] Semen Lepidii: The main active ingredients are isothiocyanate and its glycosides and flavonoids. Among them, thiocyanate and its glucosides are common components of cruciferous plants and have antitussive and antiasthmatic effects.

[0039] Almonds: The active ingredients include bitter almonds (amygadalin), almond oil (amygadalic acid), various free amino acids (amino acid), chlorogenic acid (chlorogenic acid), benzoic acid (benzoic acid), etc. Bitter almonds will decompose and produce a small amount of hydrocyanic acid after oral administration, which can inhibit the cough center and achieve antitussive and antiasthmatic effects. Almonds can also be used in a variety of clinical medicines to treat cough and asthma.

[0040] Glauber's salt: The pharmacological effects include clearing heat and promoting bowel movements, moistening dryness and softening hard masses, clearing away heat and reducing swelling, and treating sore throat, mouth ulcers, swollen and painful gums, red and swollen eyes, and red poison.

[0041] The contents of angelica dahurica, scutellaria baicalensis, scutellaria baicalensis seeds, apricot kernel and mirabilite in the traditional Chinese medicine composition can be configured according to the spread of cancer cells. The components of the traditional Chinese medicine composition are calculated by weight. The content of angelica dahurica is between 3 and 30 parts by weight, the content of scutellaria baicalensis is between 3 and 30 parts by weight, the content of scutellaria baicalensis seeds is between 8 and 35 parts by weight, the content of apricot kernel is between 5 and 30 parts by weight, and the content of mirabilite is between 0.3 and 5 parts by weight.

[0042] In a first preferred embodiment, the content configuration of the Chinese medicine composition refers to the following combination: the content of the Angelica dahurica and the Scutellaria baicalensis is a first component, the content of the Lepidium tiliaceum, the Apricot kernel and the Sodium sulfate is a second component, and the first component is greater than the second component; further illustrating that the content of the Angelica dahurica is between 5 and 10 parts by weight, the content of the Scutellaria baicalensis is between 10 and 15 parts by weight, the content of the Lepidium tiliaceum is between 10 and 15 parts by weight, the content of the Apricot kernel is between 8 and 12 parts by weight, and the content of the Sodium sulfate is between 0.5 and 1.5 parts by weight; in a preferred embodiment, the content of the Angelica dahurica and the Scutellaria baicalensis is between 10 and 15 parts by weight, the content of the Apricot kernel is between 8 and 12 parts by weight, and the content of the Sodium sulfate is between 0.5 and 1.5 parts by weight. In the embodiment, the content of Angelica dahurica is between 8 and 10 parts by weight, the content of Scutellaria baicalensis is between 13 and 15 parts by weight, the content of Lepidium tiliaceum is between 10 and 12 parts by weight, the content of Apricot kernel is between 8 and 10 parts by weight, and the content of Sodium sulfate is between 1 and 1.5 parts by weight. In actual clinical practice of traditional Chinese medicine, if the lung cancer cells in the patient's body have not spread or metastasized to other organs, the Chinese medicine composition of the first embodiment, through the specific weight parts of Angelica dahurica, Scutellaria baicalensis, Lepidium tiliaceum, Apricot kernel and Sodium sulfate and the Chinese medicine compound composition, can indeed effectively inhibit and reduce the number of active lung cancer cells in a short period of time.

[0043] In a second preferred embodiment, the configuration of the Chinese medicine composition is compared with the aforementioned first embodiment, and the content of the scutellaria baicalensis and the scutellaria leucophylla is increased, with reference to the following combination: the content of the scutellaria baicalensis and the scutellaria leucophylla is a third component, the content of the angelica dahurica, the apricot kernel and the mirabilite is a fourth component, and the third component is greater than the fourth component; further illustrating that the content of the angelica dahurica is between 5 and 10 parts by weight, the content of the scutellaria baicalensis is between 20 and 30 parts by weight, and the content of the scutellaria baicalensis is between 20 and 30 parts by weight. ~30 weight parts, the content of almond is between 8 and 12 weight parts, and the content of mirabilite is between 0.5 and 1.5 weight parts; in actual Chinese medicine clinical practice, if the lung cancer cells in the patient's body have not spread and metastasized to other organs and are combined with symptoms of pulmonary edema, the Chinese medicine composition of the second embodiment is increased by adjusting the content of the scutellaria baicalensis and the scutellaria baicalensis seeds, and after the patient regularly takes the Chinese medicine composition, it can not only inhibit and reduce the number of active lung cancer cells, but also correspondingly improve the symptoms of pulmonary edema in the patient.

[0044] In a third preferred embodiment, the configuration of the Chinese medicine composition increases the content of the Angelica dahurica, the Lemongrass seeds, the Apricot kernel and the Sodium sulfate compared to the aforementioned first embodiment, with reference to the following combination: the second component amount of the total content of the Lemongrass seeds, the Apricot kernel and the Sodium sulfate is greater than the first component amount of the total content of the Angelica dahurica and the Scutellaria baicalensis, wherein the content of the Angelica dahurica is between 15 and 25 parts by weight, the content of the Scutellaria baicalensis is between 10 and 15 parts by weight, the content of the Lemongrass seeds is between 20 and 30 parts by weight, the content of the Apricot kernel is between 15 and 25 parts by weight, and the content of the Sodium sulfate is between 2 and 4 parts by weight; in actual Chinese medicine clinical practice, if the lung cancer cells in the patient's body have spread and metastasized to other organs, the Chinese medicine composition of the third embodiment can effectively regulate and improve the spread and metastasis of cancer cells in the patient's body by adjusting the content of the Angelica dahurica, the Lemongrass seeds, the Apricot kernel and the Sodium sulfate, and the patient takes the Chinese medicine composition regularly.

[0045] In addition, the cell experiments of the Chinese medicine composition in the first to third embodiments above have confirmed that it can be used to prepare a method for inhibiting or reducing the activity of lung cancer cells. The test process of the Chinese medicine composition is to administer the lung cancer cells with an effective dose of 1 to 100 μg / ml. The test found that the effective dose of the Chinese medicine composition can effectively induce the expression of LC3-I / LC3-II protein in non-small cell lung cancer cells and small cell lung cancer cells, and trigger the autophagy of the lung cancer cells. The effective dose of the Chinese medicine composition is preferably between 25 and 100 μg / ml, and the expression of LC3-I / LC3-II protein in non-small cell lung cancer cells and small cell lung cancer cells is more significant, and the number of active lung cancer cells is significantly inhibited and reduced accordingly, thereby achieving the desired therapeutic effect of treating or improving lung cancer.

[0046] In order to fully understand the purpose, characteristics and technical effects of the present invention, the implementation content of the Chinese medicine composition described in the first embodiment is provided herewith, and different effective doses are respectively administered to non-small cell lung cancer cells and small cell lung cancer cells, wherein the non-small cell lung cancer cells described in this experiment are lung adenocarcinoma cells (H460) as cell experiments, and the small cell lung cancer cells are small cell lung cancer cells (H1355) as cell experiments, and the growth and survival rate of lung cancer cells are analyzed accordingly, thereby illustrating that the Chinese medicine composition described in this embodiment does provide the effect of inhibiting or reducing the activity of lung cancer cells.

[0047] The following specific embodiments can further demonstrate the practical application scope of the present invention, and any technician in the relevant technical field can make appropriate changes to its parameters or settings after referring to the present invention, and it should still fall within the scope of the present invention.

[0048] The groups of this experiment include control groups 1 to 3 and experimental groups:

[0049] Control group 1: Scutellaria baicalensis was used alone, and the effective doses of Scutellaria baicalensis were respectively prepared as 0μg / ml, 1μg / ml, 5μg / ml, 25μg / ml, 50μg / ml, and 100μg / ml, and each effective dose of Scutellaria baicalensis was administered to lung adenocarcinoma cells (H460) and small cell lung cancer cells (H1355), respectively.

[0050] Control group 2: Angelica dahurica was used alone, and the effective doses of the Angelica dahurica were respectively prepared as 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml, and each effective dose of the Angelica dahurica was administered to lung adenocarcinoma cells (H460) and small cell lung cancer cells (H1355).

[0051] Control group 3: The scutellaria baicalensis and the angelica dahurica were prepared in a 1:1 ratio. The effective doses of the combination of scutellaria baicalensis and the angelica dahurica were 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml. The effective doses of the combination of scutellaria baicalensis and the angelica dahurica were administered to lung adenocarcinoma cells (H460) and small cell lung cancer cells (H1355), respectively.

[0052] Experimental group: The Chinese medicine composition contained 10 parts by weight of Angelica dahurica, 15 parts by weight of Scutellaria baicalensis, 12 parts by weight of Semen Lepidii, 10 parts by weight of Apricot kernel, and 1.5 parts by weight of Sodium Sulfate. The Chinese medicine composition was formulated at 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml. The effective doses of the Chinese medicine composition were administered to lung adenocarcinoma cells (H460) and small cell lung cancer cells (H1355), respectively.

[0053] Cell viability assay (MTT):

[0054] In this experiment, 5×10 lung adenocarcinoma cells (H460) and small cell lung cancer cells (H1355) were placed in each well. 3The cells were cultured in 96-well culture dishes for 24 hours. After the lung adenocarcinoma cells (H460) and the small cell lung cancer cells (H1355) were attached to the wells, the cell culture medium was removed. Then, the effective doses of 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml and 100 μg / ml were prepared for each of the control groups 1 to 3 and the experimental group, respectively, and injected into the wells corresponding to the lung adenocarcinoma cells (H460) and the small cell lung cancer cells (H1355), so that the different effective doses of each of the control groups 1 to 3 and the experimental group were respectively injected into the wells corresponding to the lung adenocarcinoma cells (H460) and the small cell lung cancer cells (H1355). The cells were treated with MTT in the wells of the lung adenocarcinoma cells (H460) and the small cell lung cancer cells (H1355) for 1 day and 2 days, and then the culture medium was removed, and the cells were washed twice with sterile phosphate buffered saline (PBS), and 0.2 ml of MTT culture medium was added to the wells of the lung adenocarcinoma cells (H460) and the small cell lung cancer cells (H1355) corresponding to the control groups 1 to 3 and the experimental groups, and placed in a constant temperature incubator for 2 hours, and the MTT culture medium was removed, and the cells were washed once with non-sterile phosphate buffered saline (PBS), and finally 0.2 ml of DMSO was added to dissolve the crystals, and the optical density (OD) value was measured at a wavelength of 570 nm using an enzyme immunoassay (ELISA reader) analyzer to calculate the relative cell survival rate.

[0055] The test data of each of the control groups 1 to 3 are based on 0 μg / ml without drug treatment as the control group, and the absorbance value measured in the control group is quantified as the percentage of 100%, and then the OD value of each of the control groups 1 to 3 is divided by the control group to calculate the relative lung adenocarcinoma cell (H460) survival rate percentage and the small cell lung cancer cell (H1355) survival rate percentage; the test data of the experimental group are based on 0 μg / ml without drug treatment as the control group, and the absorbance value measured in the control group is quantified as the percentage of 100%, and then the OD value of the experimental group is divided by the control group to calculate the relative lung adenocarcinoma cell (H460) survival rate percentage and the small cell lung cancer cell (H1355) survival rate percentage.

[0056] Autophagy cell function test - Western blot:

[0057] Detection of the expression of autophagy corresponding to antibodies LC3-I, LC3-II and B-Actin. First, the nitrocellulose filter membrane (NC) and the antibodies were incubated in phosphate buffered saline (PBS) containing 2.5% bovine serum albumin (BSA) for 3 hours, and then rinsed with phosphate buffered saline (PBS) for 3 times, and then horse-radish peroxidase (HRP) antibody was added and incubated for another hour. Finally, the antigen-antibody complex was detected with Immobilion Western HRP enzyme cold light reagent, and the protein blotting was quantified using an optical density instrument (Appraise, Beckman-Coulter, Brea, California, USA).

[0058] In particular, autophagy is a physiological function in cells, which mainly decomposes old proteins in cells, metabolizes ineffective organelles and foreign microorganisms. There are many proteins involved in the regulation of autophagy. Microtubule-associated protein 1A / 1B-light chain 3 (LC3 for short) located in the cytoplasm is often used as a fluorescent marker for autophagy activation. When autophagy is activated, LC3 will combine with lipid phosphatidyl-ethanolamine to form LC3-II, so that LC3-II is embedded in the membrane of the phagocytic vesicle. Therefore, the amount of LC3-II generated in the vesicle can be used as an activation indicator of autophagy.

[0059] 1. To investigate the cell survival rate and I1057TW of the lung adenocarcinoma cells (H460) in response to the administration of the control groups 1 to 3 and the experimental groups.

[0060] Autophagy:

[0061] 1. The cell viability test of the lung adenocarcinoma cells (H460): analyzing the survival status of the lung adenocarcinoma cells (H460) corresponding to the control groups 1 to 3 and the experimental groups, wherein the control groups 1 to 3 and the experimental groups are respectively detected at different effective doses of the control groups 1 to 3 and the experimental groups after administration of the lung adenocarcinoma cells (H460), corresponding to the cell viability of the lung adenocarcinoma cells (H460) on the first day and the second day after administration.

[0062] The observation results are as follows Figure 1A , Figure 2A , Figure 3A and Figure 4A,in Figure 1A The cell survival rate data of lung adenocarcinoma cells (H460) corresponding to the control group 1 on the first day and the second day after administration of different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml; Figure 2A The cell survival rate data of lung adenocarcinoma cells (H460) corresponding to the control group 2 on the first day and the second day after administration of different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml; Figure 3A The cell survival rate data of lung adenocarcinoma cells (H460) corresponding to the control group 3 on the first day and the second day after administration of different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml; Figure 4A The figures are the cell survival rate data of lung adenocarcinoma cells (H460) on day 1 and day 2 corresponding to the experimental group when different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml were administered.

[0063] Figure 1A , Figure 2A , Figure 3A and Figure 4A The standard error (SE) data results of the control groups 1 to 3 and the experimental groups are shown, which are the detection values ​​of three independent experiments (n=3) performed on each group of cell tests, wherein "*" indicates a comparison of the values ​​of the control groups 1 to 3 and the experimental group, p<0.05.

[0064] like Figure 1A As shown, from the analysis of the control group 1 on the first day, it was found that the cell survival rate of the lung adenocarcinoma cells (H460) corresponding to different effective doses of the control group 1 did not decrease significantly; from the analysis of the control group 1 on the second day, it was found that the cell survival rate of the lung adenocarcinoma cells (H460) corresponding to the effective dose of 100 μg / ml of the control group 1 significantly decreased to about 50%, indicating that the control group 1 using only Scutellaria baicalensis at an effective dose of 100 μg / ml has a certain ability to inhibit the activity of lung adenocarcinoma cells (H460) and reduce the number of lung adenocarcinoma cells (H460).

[0065] like Figure 2AAs shown, the analysis of the control group 2 on the first day and the second day showed that the cell survival rate of the lung adenocarcinoma cells (H460) corresponding to different effective doses of the control group 2 did not decrease significantly, indicating that the control group 2, which was administered Angelica dahurica alone, could not provide the ability to effectively inhibit and reduce the activity of lung adenocarcinoma cells (H460).

[0066] like Figure 3A As shown, from the analysis of the control group 3 on the first day, it was found that the cell survival rate of the lung adenocarcinoma cells (H460) corresponding to different effective doses of the control group 3 did not decrease significantly; from the analysis of the control group 3 on the second day, it was found that the cell survival rate of the lung adenocarcinoma cells (H460) corresponding to the effective dose of 100 μg / ml of the control group 3 significantly decreased to about 20-30%, that is, the control group 3 using the combination of Scutellaria baicalensis and Angelica dahurica was compared with the control group 1 using Scutellaria baicalensis alone, and the control group 3 more significantly inhibited the activity of lung adenocarcinoma cells (H460), and correspondingly reduced the number of lung adenocarcinoma cells (H460).

[0067] like Figure 4A As shown, from the analysis of the experimental group on the first day, it was found that the cell survival rate of the lung adenocarcinoma cells (H460) in the experimental group at the effective doses of 25 μg / ml, 50 μg / ml and 100 μg / ml was significantly reduced to about 5-20%, wherein the cell survival rate of the lung adenocarcinoma cells (H460) in the experimental group at the effective dose of 100 μg / ml was significantly reduced; from the analysis of the experimental group on the second day, it was found that the cell survival rate of the lung adenocarcinoma cells (H460) in the experimental group at the effective doses of 25 μg / ml, 50 μg / ml and 100 μg / ml was significantly reduced to about 5-20%. The experimental group showed a more significant decrease in the cell survival rate of the lung adenocarcinoma cells (H460) at an effective dose of 100 μg / ml, that is, the Chinese medicine composition of the experimental group, compared with the control group 3 using the combination of the scutellaria baicalensis and the angelica dahurica, more significantly inhibited the activity of the lung adenocarcinoma cells (H460) and correspondingly reduced the number of the lung adenocarcinoma cells (H460). When the experimental group was administered with effective doses of 25 μg / ml, 50 μg / ml and 100 μg / ml on the first day, the efficacy of significantly reducing the number of the lung adenocarcinoma cells (H460) was obtained.

[0068] 2. Autophagy ability of the lung adenocarcinoma cells (H460): The control groups 1 to 3 and the experimental groups were respectively injected with effective doses of 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml and 100 μg / ml to analyze the expression of LC3-I / LC3-II proteins in the corresponding lung adenocarcinoma cells (H460). The control groups 1 to 3 and the experimental groups were respectively subjected to electrophoresis to detect the protein expression of LC3-I / LC3-II in the lung adenocarcinoma cells (H460) after administration of the lung adenocarcinoma cells (H460).

[0069] The observation results are as follows Figure 1B , Figure 2B , Figure 3B and Figure 4B ,in Figure 1B The electrophoresis of LC3-I / LC3-II proteins in lung adenocarcinoma cells (H460) corresponding to the control group 1 when different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml were administered; Figure 2B The electrophoresis of LC3-I / LC3-II proteins in lung adenocarcinoma cells (H460) corresponding to the control group 2 when different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml were administered; Figure 3B The electrophoresis of LC3-I / LC3-II proteins in lung adenocarcinoma cells (H460) corresponding to the control group 3 when different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml were administered; Figure 4B The electrophoresis of LC3-I / LC3-II proteins in the corresponding lung adenocarcinoma cells (H460) when the experimental groups were administered with different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml.

[0070] like Figure 1B , Figure 2B , Figure 3B and Figure 4BAs shown, the experimental group at effective doses of 25 μg / ml, 50 μg / ml and 100 μg / ml can significantly increase the LC3-II protein expression in the lung adenocarcinoma cells (H460) compared to the control groups 1 to 3, indicating that the experimental group at effective doses of 25 μg / ml, 50 μg / ml and 100 μg / ml can significantly induce the protein expression of LC3-I / LC3-II in the lung adenocarcinoma cells (H460), thereby triggering the autophagy of the lung adenocarcinoma cells (H460), so as to achieve the effect of effectively inhibiting and reducing the activity of the lung adenocarcinoma cells (H460).

[0071] 2. To investigate the cell survival rate and autophagy of the small cell lung cancer cells (H1355) administered to the control groups 1 to 3 and the experimental groups:

[0072] 1. Cell viability test of the small cell lung cancer cells (H1355): analyzing the survival status of the small cell lung cancer cells (H1355) corresponding to the control groups 1 to 3 and the experimental groups, wherein the control groups 1 to 3 and the experimental groups are respectively detected at different effective doses of the control groups 1 to 3 and the experimental groups after the small cell lung cancer cells (H1355) are administered, and the cell viability of the small cell lung cancer cells (H1355) on the first day and the second day after the administration is detected.

[0073] The observation results are as follows Figure 5A , Fig. 6A , Fig. 7A and Fig. 8A ,in Figure 5A The cell survival rate data of the control group 1 on the first day and the second day corresponding to the small cell lung cancer cells (H1355) administered with different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml; Fig. 6A The cell survival rate data of the small cell lung cancer cells (H1355) corresponding to the control group 2 on the first day and the second day after administration of different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml; Fig. 7A The cell survival rate data of the small cell lung cancer cells (H1355) corresponding to the control group 3 on the first day and the second day after administration of different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml; Fig. 8AThe cell survival rate data of small cell lung cancer cells (H1355) on day 1 and day 2 corresponding to the experimental group when different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml were administered.

[0074] Figure 5A , Fig. 6A , Fig. 7A and Fig. 8A The standard error (SE) data results of the control groups 1 to 3 and the experimental groups are shown, which are the detection values ​​of three independent experiments (n=3) performed on each group of cell tests, wherein "*" indicates a comparison of the values ​​of the control groups 1 to 3 and the experimental group, p<0.05.

[0075] like Figure 5A As shown, the analysis of the control group 1 on the first day and the second day showed that the cell survival rate of the small cell lung cancer cells (H1355) corresponding to different effective doses of the control group 1 did not decrease significantly, indicating that the control group 1, which only administered Scutellaria baicalensis, could not provide effective inhibition and reduction of the number of active small cell lung cancer cells (H1355).

[0076] like Fig. 6A As shown, the analysis of the control group 2 on the first and second days revealed that the cell survival rates of the small cell lung cancer cells (H1355) corresponding to different effective doses of the control group 2 did not decrease significantly, indicating that the administration of Angelica dahurica alone to the control group 2 was the same as that of the control group 1, that is, it could not provide effective inhibition and reduction of the activity of small cell lung cancer cells (H1355).

[0077] like Fig. 7A As shown, the analysis of the control group 3 on the first and second days revealed that the cell survival rates of the small cell lung cancer cells (H1355) corresponding to different effective doses of the control group 3 did not decrease significantly, indicating that the combination of Scutellaria baicalensis and Angelica dahurica used in the control group 3 was the same as that of the control groups 1 and 2, that is, it could not provide the efficacy of effectively inhibiting and reducing the activity of the small cell lung cancer cells (H1355).

[0078] like Fig. 8AAs shown, from the analysis of the experimental group on the first day, it was found that the cell survival rate of the small cell lung cancer cells (H1355) in the experimental group at the effective doses of 25 μg / ml, 50 μg / ml and 100 μg / ml was significantly reduced to about 20-60%, wherein the experimental group at the effective dose of 100 μg / ml showed a decreased cell survival rate of the small cell lung cancer cells (H1355); from the analysis of the experimental group on the second day, it was found that the cell survival rate of the small cell lung cancer cells (H1355) in the experimental group at the effective doses of 25 μg / ml, 50 μg / ml and 100 μg / ml was significantly reduced to about 20-60%. The survival rate was significantly reduced to about 10-50%. Similarly, the experimental group had a more significant reduction in the cell survival rate of the small cell lung cancer cells (H1355) at an effective dose of 100 μg / ml. That is, the Chinese medicine composition of the experimental group significantly inhibited the activity of small cell lung cancer cells (H1355) compared with the control groups 1-3, and correspondingly reduced the number of small cell lung cancer cells (H1355). When the experimental group was administered with effective doses of 25 μg / ml, 50 μg / ml and 100 μg / ml on the first day, the efficacy of significantly reducing the number of small cell lung cancer cells (H1355) was obtained.

[0079] 2. Autophagy ability of the small cell lung cancer cells (H1355): Analyze the expression of LC3-I / LC3-II proteins in the small cell lung cancer cells (H1355) corresponding to the control groups 1 to 3 and the experimental groups, wherein the control groups 1 to 3 and the experimental groups are respectively detected at different effective doses of the control groups 1 to 3 and the experimental groups after the small cell lung cancer cells (H1355) are administered, and the expression of LC3-I / LC3-II proteins in the corresponding small cell lung cancer cells (H1355) is detected by electrophoresis.

[0080] The observation results are as follows Figure 5B , Figure 6B , Figure 7B and Figure 8B ,in Figure 5B The LC3-I / LC3-II protein electrophoresis of the small cell lung cancer cells (H1355) corresponding to the control group 1 when different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml were administered; Figure 6B The LC3-I / LC3-II protein electrophoresis of the small cell lung cancer cells (H1355) corresponding to the control group 2 when different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml were administered; Figure 7BThe LC3-I / LC3-II protein electrophoresis of the small cell lung cancer cells (H1355) corresponding to the control group 3 when different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml were administered; Figure 8B The LC3-I / LC3-II protein electrophoresis of the corresponding small cell lung cancer cells (H1355) in the experimental group after administration of different effective doses of 0 μg / ml, 1 μg / ml, 5 μg / ml, 25 μg / ml, 50 μg / ml, and 100 μg / ml.

[0081] like Figure 5B , Figure 6B , Figure 7B and Figure 8B As shown, the experimental group at an effective dose of 50 μg / ml and 100 μg / ml can significantly increase the LC3-II protein expression in the small cell lung cancer cells (H1355) compared to the control groups 1 to 3, indicating that the experimental group at an effective dose of 50 μg / ml and 100 μg / ml can significantly induce the protein expression of LC3-I / LC3-II in the small cell lung cancer cells (H1355), thereby triggering the autophagy of the small cell lung cancer cells (H1355), so as to achieve the effect of effectively inhibiting and reducing the activity of the small cell lung cancer cells (H1355).

[0082] Summarizing the above experiments, the Chinese medicine composition described in this embodiment is composed of specific weight portions of Angelica dahurica, Scutellaria baicalensis, Lemongrass seeds, Apricot kernel and Sodium sulfate and a Chinese medicine compound. Cell experiments have confirmed that the pharmacological addition and subtraction effects of Angelica dahurica, Scutellaria baicalensis, Lemongrass seeds, Apricot kernel and Sodium sulfate in the Chinese medicine composition, compared with the use of Angelica dahurica or Scutellaria baicalensis alone, and the combination of Angelica dahurica and Scutellaria baicalensis, the Chinese medicine composition can effectively inhibit the activity of lung adenocarcinoma cells (H460) and small cell lung cancer cells (H1355) in a short time when applied to a specific effective dose, and correspondingly reduce the number of lung adenocarcinoma cells (H460) and small cell lung cancer cells (H1355), wherein the Chinese medicine composition is applied at 25 μg / ml , 50 μg / ml and 100 μg / ml have the effect of preferably inhibiting and reducing the number of active lung adenocarcinoma cells (H460) and small cell lung cancer cells (H1355). In a preferred embodiment, the effective doses of 50 μg / ml and 100 μg / ml of the Chinese medicine composition have the best effect of inhibiting and reducing the number of active lung adenocarcinoma cells (H460) and small cell lung cancer cells (H1355), and can effectively induce the protein expression of LC3-I / LC3-II in the lung adenocarcinoma cells (H460) and small cell lung cancer cells (H1355), thereby triggering the lung adenocarcinoma cells (H460) and small cell lung cancer cells (H1355) to produce autophagy, thereby achieving the desired therapeutic effect of treating or improving lung cancer.

[0083] The above description is only a preferred feasible embodiment of the present invention and is not intended to limit the present invention. A person skilled in the art of the present invention, after referring to the above teachings, should be able to make appropriate modifications to the contents of the above embodiments or implement the technical effects claimed by the present invention according to different experimental methods.

Claims

1. A Chinese medicine composition for treating lung cancer, comprising Angelica dahurica, Scutellaria baicalensis, Lemongrass seeds, Apricot kernel and Sodium sulfate, wherein the content of the Chinese medicine composition is in parts by weight: 3 to 30 parts by weight of Angelica dahurica, 3 to 30 parts by weight of Scutellaria baicalensis, 8 to 35 parts by weight of Lemongrass seeds, 5 to 30 parts by weight of Apricot kernel and 0.3 to 5 parts by weight of Sodium sulfate.

2. The Chinese medicine composition for treating lung cancer as claimed in claim 1, wherein the total content of the Angelica dahurica and the Scutellaria baicalensis is a first component amount, the total content of the Lemongrass seeds, the Apricot kernel and the Sodium Sulfate is a second component amount, and the first component amount is greater than the second component amount.

3. The Chinese medicine composition for treating lung cancer as claimed in claim 2, wherein the content of Angelica dahurica is between 5 and 10 parts by weight, the content of Scutellaria baicalensis is between 10 and 15 parts by weight, the content of Lemongrass seeds is between 10 and 15 parts by weight, the content of Apricot kernel is between 8 and 12 parts by weight, and the content of Sodium sulfate is between 0.5 and 1.5 parts by weight.

4. The Chinese medicine composition for treating lung cancer as claimed in claim 3, wherein the content of Angelica dahurica is between 8 and 10 parts by weight, the content of Scutellaria baicalensis is between 13 and 15 parts by weight, the content of Lemongrass seeds is between 10 and 12 parts by weight, the content of Apricot kernel is between 8 and 10 parts by weight, and the content of Sodium sulfate is between 1 and 1.5 parts by weight.

5. The Chinese medicine composition for treating lung cancer as claimed in claim 1, wherein the total content of the scutellaria baicalensis and the scutellaria baicalensis seeds is a third component amount, the total content of the angelica dahurica, the apricot kernel and the mirabilite is a fourth component amount, and the third component amount is greater than the fourth component amount.

6. The Chinese medicine composition for treating lung cancer as claimed in claim 5, wherein the content of Angelica dahurica is between 5 and 10 parts by weight, the content of Scutellaria baicalensis is between 20 and 30 parts by weight, the content of Lemongrass seeds is between 20 and 30 parts by weight, the content of Apricot kernel is between 8 and 12 parts by weight, and the content of Sodium sulfate is between 0.5 and 1.5 parts by weight.

7. The Chinese medicine composition for treating lung cancer as claimed in claim 1, wherein the total content of the Angelica dahurica and the Scutellaria baicalensis is a fifth component amount, the total content of the Lemongrass seeds, the Apricot kernel and the Sodium Sulfate is a sixth component amount, and the amount of the sixth component is greater than the amount of the fifth component.

8. The Chinese medicine composition for treating lung cancer as claimed in claim 7, wherein the content of Angelica dahurica is between 15 and 25 parts by weight, the content of Scutellaria baicalensis is between 10 and 15 parts by weight, the content of Lemongrass seeds is between 20 and 30 parts by weight, the content of Apricot kernel is between 15 and 25 parts by weight, and the content of Sodium sulfate is between 2 and 4 parts by weight.

9. A use of a Chinese medicine composition as claimed in any one of claims 1 to 8 for preparing a method for inhibiting or reducing the activity of lung cancer cells, wherein the Chinese medicine composition is administered to the lung cancer cells at an effective dose of 1 μg / ml to 100 μg / ml to induce the protein expression of LC3-I / LC3-II and trigger the autophagy of the lung cancer cells.

10. Use of the Chinese medicine composition according to claim 9 for preparing a method for inhibiting or reducing the activity of lung cancer cells, wherein the effective dose of the Chinese medicine composition is between 25 μg / ml and 100 μg / ml, and the lung cancer cells comprise non-small cell lung cancer cells or small cell lung cancer cells.

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