CN114558000B - Application of sesamin in preparation of medicine for treating melanoma - Google Patents

Abstract

The invention relates to application of praeruptorin in preparing a medicine for treating melanoma. The invention discloses the effect of the white flower falcarind in the aspect of treating melanoma, and experimental results show that the white flower falcarind has obvious anti-tumor effect on in-vivo melanoma cells, can effectively enhance the activities of cytokines and NK cells in serum of tumor-bearing mice, and indicate that the anti-tumor effect can be related to enhancing immunity. Can be used for preparing anti-melanoma medicine.

Description

Application of sesamin in preparation of medicine for treating melanoma

Technical Field

The invention relates to the field of medicines, in particular to an application of praeruptorin in preparation of a medicine for treating melanoma.

Background

Human melanoma, particularly malignant melanoma, is a highly metastatic tumor that is markedly resistant to chemotherapy. After decades of treatment and research, the survival rate of melanoma patients is 3% -5%, especially in adult primary tumors, increases more rapidly. Therefore, searching for high-efficiency low-toxicity traditional Chinese medicines to change the biological behaviors of tumors has become an important strategy for tumor treatment.

Peucedanin (Praerupterin B, CAS number 81740-07-0; molecular formula C) ₂₄ H ₂₆ O ₇ The method comprises the steps of carrying out a first treatment on the surface of the Molecular weight: 426.46 Is an angular pyranocoumarin component extracted from radix Peucedani, and has effects of reducing qi phlegm, dispelling pathogenic wind and clearing heat. In recent years, it has been found that the compounds can improve diet-induced hyperlipidemia and alleviate insulin resistance by modulating SREBP signaling pathways. However, no report on the treatment of melanoma by the compound is currently available.

Disclosure of Invention

The invention provides a method for filling the blank of the prior art, firstly discloses the pharmaceutical activity of the praeruptorin in treating melanoma, and provides the following technical scheme:

in a first aspect of the invention, there is provided the use of peucedanum praeruptorum as active ingredient in the manufacture of a medicament for the treatment of melanoma.

In one embodiment, the use comprises inhibiting melanoma growth.

In one embodiment, the use includes increasing IFN-gamma expression levels and decreasing IL-4 secretion levels in serum.

In one embodiment, the use comprises enhancing NK cell killing activity in vivo.

In one embodiment, the medicament is a compound preparation prepared by compounding the peucedanum praeruptorum and auxiliary materials conventional in the art.

In one embodiment, the compound preparation is in the form of a tablet, granule, pill, capsule or injection.

Detailed Description

The following description of the preferred embodiments of the present invention is provided for the purpose of illustration and explanation only and is not intended to limit the present invention.

EXAMPLE 1 tumor-inhibiting effect of Peucedanin

1. Experimental materials: the praeruptorin (hereinafter referred to as PB) is purchased from the biological technology Co., ltd. Of Dou Geli (purity not less than 98%); thiazole blue (MTT) was purchased from Amersco, USA, RPMI1640 medium, trypsin, fetal bovine serum was purchased from Gibco, USA, and mouse IFN-gamma and IL-4 detection kits were purchased from Nanjing Diels Biotechnology Co.

2. Experimental cells and animals:

human melanoma a375 cell line was purchased from Shanghai Bomaide biotechnology Co., ltd; BALB/c mice, 18-20g in weight, were purchased from Peking Vitre Liwa laboratory animal technologies Co.

3. The experimental method and the grouping are as follows:

after the A375 cells growing in logarithmic phase are digested with pancreatin, the cells are centrifuged to prepare a cell suspension, and the right side of the mice is inoculated with about 2X 10 injections ⁵ Cells/cells. After 3 weeks, mice with successfully modeled transplants (diameter not less than 5 mm) were screened, and the transplants were further divided into 5 groups, i.e., physiological saline group (0.9% sodium chloride injection, NS control group), acacia control group (5% acacia aqueous solution), PB low dose group (5 mg/(kg·d)), PB medium dose group (10 mg/(kg·d)) and PB high dose group (20 mg/(kg·d)), 10 per group, by adopting a random number table method. PB and acacia were suspended, and each mouse was perfused with either PB or NS at 0.5ml/d for 28d (culling if tumor length > 17 mm), then the mice were sacrificed and the tumor mass was weighed and tumor inhibition was calculated (tumor growth inhibition rate, TGI).

TGI calculation formula:

TGI (%) = (average mass of experimental control group graft versus average mass of experimental group graft)/average mass of experimental control group graft x 100%

Statistical methods: all data are analyzed by adopting SPSS 17.0 statistical software, metering data are expressed by mean ± standard deviation, single-factor analysis of variance is carried out among groups, and P < 0.05 is taken as a difference to have statistical significance.

4. Experimental results:

compared with normal mice, the food intake and water intake of tumor-bearing mice are reduced, the hair color becomes dry and yellow, and the hair is partially dehaired and has poor mental condition. After 28d of administration, the PB treated mice had improved mental status and gradually restored their hair gloss, with an increase in dietary intake. As shown in Table 1, the difference in mass of mice in each of the groups of the transplanted tumor models before administration was not statistically significant, and P was > 0.05. Mice became lighter in body mass after PB treatment, with differences statistically significant, p=0.05, relative to NS control; wherein the mass of the medium dose group body is reduced to the minimum, and P is less than 0.01. The cytoma mass is compared, and compared with an NS control group, the difference of each experimental group has statistical significance, and P is less than 0.05; wherein the difference is most pronounced in the high dose group, P < 0.01.

Table 1 PB effect on mouse body weight and tumor mass (n=10)

Note that: compared with the NS control group, the P is less than 0.05, and the #P is less than 0.01

EXAMPLE 2 Effect of Peucedanin on mouse IFN-gamma and IL-4 secretion levels

ELISA method for detecting IFN-gamma and IL-4 expression in serum

The procedure is described in detail with reference to IFN-gamma and IL-4 detection kits: taking a kit and standing at room temperature, coating a plate with a negative control hole, 8 standard substance holes (2000, 1000, 500, 250, 125, 62.5, 31.25 and 15.625 ng/L) with different concentrations for detecting IFN-gamma, 8 standard substance holes (500, 250, 125, 62.5, 31.25 and 15.625, 7.8125 and 3.91 pg/L) with different concentrations for detecting IL-4, directly adding 50 mu L of sample diluent into the negative hole, and adding 40 mu L of sample diluent into the rest holes and 10 mu L of sample; incubating for 60min at 37deg.C with sealing plate membrane, discarding supernatant, washing, repeating for 5 times, and drying; adding 50 mu L of enzyme-labeled reagent into each hole, continuously incubating for 40min, discarding supernatant, washing with washing solution for 5 times, and drying; 50 mu L of each of the color developing agents A and B is respectively added into each hole, and the mixture is gently mixed, and the color is developed for 10min at 37 ℃ in a dark place; the color development was stopped by adding 50. Mu.L of stop solution to each well. Absorbance (a value) of each well was measured at a wavelength of a blank Kong Diaoling, 450nm and a corresponding concentration value was calculated.

2. Experimental results

As shown in Table 2, PB enhances IFN-gamma expression level in serum of experimental mice, and compared with NS control group, the difference of each group of PB experiment has statistical significance, and P is less than 0.05; wherein, the secretion level of the high-dose group is highest and reaches 721.98 +/-12.47 ng/L, and P is less than 0.01. While the bleeding level of IL-4 in each group is reduced by PB treatment, the reduction of the high-dose group is most obvious, and the dosage reaches 120.68 +/-5.26 ng/L and P is less than 0.05.

Table 2 PB effect on IFN-. Gamma.and IL-4 expression in serum of tumor-bearing mice (n=10)

Note that: compared with the NS control group, the P is less than 0.05, and the #P is less than 0.01

EXAMPLE 3 Effect of Peucedanin on NK cell killing Activity of transplanted tumor mice

Determination of NK cell killing Activity of mice by MTT method

1.1 preparation of potent and target cells mice were taken from spleen and inguinal lymph nodes and made into lymphocyte suspensions using RP containing 10% calf serum as effector cellsMI1640 culture medium was adjusted to 1X 10 cell concentration ⁶ mL ^-1 、5×10 ⁵ mL ^-1 And 2X 10 ⁵ mL ^-1 . The target cells are YAC-1 cells, and the cell concentration is adjusted to be 1 multiplied by 10 ⁴ mL ^-1 。

1.2 NK cell killing activity assay in 96 well plates, assay Kong Jiaxiao, 50. Mu.L each of target cells; effector control wells plus 50 μl of effector and 1640 medium each; target cells Kong Jiaba and 1640 were each 50. Mu.L in culture. 3 compound wells are made for each specimen, and the effective target ratios are 100:1, 50:1 and 20:1 respectively. Placing the culture plate into CO ₂ Culturing in an incubator for 20h, and adding 20 mu L of 5mg/ml MTT into each well for further incubation for 4h; after completion, 150. Mu.L of dimethyl sulfoxide (DMSO) was added to each well to dissolve the precipitate, and the mixture was subjected to shaking by a micro-oscillator for 10 minutes, and absorbance (A value) of each well was measured at a wavelength of 490nm on an microplate reader.

Killing activity (%) = (1- (experimental group mean a value-effector group mean a value)/target cell group mean a value) ×100%

2. Experimental results

As shown in Table 3, PB can enhance the killing activity of NK cells of mice, the increase of the killing activity of NK cells is obvious when the effective target ratio is 50:1, the killing activity of NK cells of each treatment group of PB is higher than that of an NS control group, the difference is statistically significant, and P is less than 0.05; and the NK cell activity of the high-dose group reaches 50.32%, the increase is most obvious, and P is less than 0.01.

Table 3 PB effect on NK cell activity of tumor-bearing mice (n=10)

Note that: compared with the NS control group, the P is less than 0.05, and the #P is less than 0.01

It will be apparent to those skilled in the art that various modifications and variations can be made to the present invention without departing from the spirit or scope of the invention. Thus, it is intended that the present invention also include such modifications and alterations insofar as they come within the scope of the appended claims or the equivalents thereof.

Claims (3)

1. Application of sesamin as active ingredient in preparing medicine for treating melanoma is provided.

2. The use according to claim 1, wherein the medicament is a pharmaceutical formulation of praeruptorin in combination with adjuvants conventional in the art.

3. The use according to claim 2, wherein the pharmaceutical formulation is in the form of a tablet, granule, pill, capsule or injection.