CN114028417A - Application of dendrobium nobile lindl polysaccharide in preparation of medicine for treating retinoblastoma - Google Patents

Abstract

The application discloses application of dendrobium nobile polysaccharide in preparation of a medicine for treating retinoblastoma in the technical field of antitumor drugs, the dendrobium nobile polysaccharide is obtained after fresh stem extraction and alcohol precipitation, and the result shows that dendrobium nobile polysaccharide with different concentrations has significant tumor killing activity on retinoblastoma, and the tumor killing activity becomes concentration dependence, so that the dendrobium nobile polysaccharide has very good medicine research and development potential for treating retinoblastoma.

Description

Application of dendrobium nobile lindl polysaccharide in preparation of medicine for treating retinoblastoma

Technical Field

The invention relates to the technical field of antitumor drugs, and in particular relates to application of dendrobium nobile lindl polysaccharide in preparation of a drug for treating retinoblastoma.

Background

Retinoblastoma (Rb) is a malignant tumor derived from photoreceptor precursor cells. The traditional Chinese medicine is a physical treatment method which has obvious effect, is expensive and complex in operation, is difficult to accurately act on eyeground in the aspect of medicine application, and is difficult to accurately act on the eyeground due to the fact that the diameter of a capillary at the retina is very thin and is positioned at the tail end of a blood circulation system in vivo. The cytotoxic drugs are the mainstream chemotherapy method for various solid tumors of children at present; chemotherapy is limited to vincristine, carboplatin and cyclophosphamide, which are traditional cytotoxic drugs, and Vincristine (VCR) is the most commonly used, and the microtubule inhibitor vincristine has a broad-spectrum anti-tumor effect but has high toxic and side effects, so that a drug with higher safety and higher inhibition effect is urgently needed to be found.

Dendrobium nobile lindl is a fresh or dry stem of a plant of the genus Dendrobium of the family Orchidaceae, is mainly produced in Yunnan, Guizhou, Sichuan, Guangxi, Henan Funiu mountain and the like in China, and is a commonly used famous and precious Chinese medicinal material. The chemical components mainly comprise polysaccharide, alkaloid, bibenzyl, phenanthrene, sesquiterpene glucoside, volatile oil and steroid. The polysaccharide is one of the main components, has the functions of enhancing the immunity of the organism, reducing blood sugar, resisting tumor, resisting aging, resisting oxidation and the like, and has higher medicinal value.

In the research of the prior art, the dendrobium nobile lindl polysaccharide shows higher inhibition to S180 tumor cells (ascites tumor cells) and shows stronger resistance to the multiplication of human promyelocytic leukemia cells in vitro. However, the inhibitory activity of dendrobium nobile lindl polysaccharide on retinoblastoma is not reported at present.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides the application of dendrobium nobile lindl polysaccharide in preparing the medicine for treating retinoblastoma.

One of the purposes of the invention is to provide the application of dendrobium nobile lindl polysaccharide in the preparation of the medicine for treating retinoblastoma.

Specifically, the dendrobium nobile polysaccharide is prepared by the following steps:

step one, crushing: taking fresh dendrobium stem strips, removing leaves, cleaning and crushing for later use;

step two, adding 2-6 times of water into the fragments obtained in the step one according to the mass ratio, soaking for a period of time, filtering to obtain filter residues and filtrate, and repeating the soaking and filtering operation on the filter residues for 1-4 times to obtain 2-5 times of filtrate; step three, combining the filtrates, carrying out vacuum rotary evaporation and concentration to obtain a concentrated solution, centrifuging, and keeping a supernatant;

step four, adding 3-5 times of volume of absolute ethyl alcohol into the concentrated solution obtained in the step three to carry out alcohol precipitation;

and fifthly, centrifuging the solution precipitated in the fourth step, removing the supernatant, dissolving the precipitate with water, then carrying out vacuum rotary evaporation concentration, and carrying out freeze drying to obtain the dendrobium nobile polysaccharide.

Experiments show that the dendrobium nobile lindl polysaccharide has obvious tumor killing activity on retinoblastoma, and the tumor killing activity is concentration-dependent.

Drawings

FIG. 1 is a graph showing the comparison of the inhibitory activity of Dendrobium nobile Lindl polysaccharides against retinoblastoma at different concentrations.

Detailed Description

The following is further detailed by way of specific embodiments:

1. preparation of dendrobium nobile polysaccharide

Taking 500 g of fresh dendrobium stem strips, removing leaves, cleaning, cutting, crushing in a crusher, adding 1 liter of water into the crushed dendrobium stem fragments, soaking for 60 minutes, filtering with 500-mesh gauze, adding 1 liter of water into filter residues, continuously soaking for 60 minutes, filtering with 500-mesh gauze, adding 1 liter of water into the filter residues, soaking for 60 minutes, and filtering with 500-mesh gauze.

And combining the three filtrates, concentrating by vacuum rotary evaporation at 70 ℃ until the volume is 500 ml, centrifuging at 5000rpm for 10 minutes, adding 2L of absolute ethyl alcohol into the supernatant, precipitating with ethanol overnight, centrifuging at 5000rpm for 10 minutes, discarding the supernatant, adding 1L of distilled water into the precipitate, dissolving by vacuum rotary evaporation at 70 ℃ until the volume is 200 ml, and freeze-drying by a freeze-dryer to obtain 12.64 g of the dendrobium nobile polysaccharide (the polysaccharide yield is 2.53%).

2. Inhibition activity assay

2.1 collecting RB-Y79 (human retinoblastoma) log phase cells, adjusting the concentration of RB-Y79 cell suspension to adjust the density of RB-Y79 cells to 3000 cells/well, adding 100. mu.L of cell suspension per well and spreading in 96-well plates (marginal wells filled with sterile PBS). 5% CO₂Incubation was performed at 37 ℃ for about 12 h.

2.2, adding medicine after culturing cells for 12h, taking dendrobium nobile polysaccharide, setting 5 medicine concentrations, namely 0, 0.5, 1, 2 and 5mg/mL, for treatment, setting 5 compound holes in each hole with 100 mu L, and continuing to incubate for 72 h.

2.3, adding 20 mu L of CCK-8 solution into each well under the condition of keeping out of the light, and continuing culturing for 4 hours.

And 2.4, after culturing for 4h, measuring the light absorption value of each hole at 450nm of an enzyme-linked immunosorbent assay detector under a dark condition, and analyzing the significant difference between each drug treatment group and a control group by using 0mg/mL as the control group through single-factor variance analysis.

2.5, as shown in fig. 1, the results show that the dendrobium nobile lindl polysaccharides with different concentrations have significant tumor killing activity on retinoblastoma, and the tumor killing activity is concentration-dependent, so that the dendrobium nobile lindl polysaccharides have very good drug development potential for treating retinoblastoma.

The foregoing is merely an example of the present invention and common general knowledge of known specific structures and features of the embodiments is not described herein in any greater detail. It should be noted that, for those skilled in the art, without departing from the structure of the present invention, several changes and modifications can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicability of the patent. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.

Claims (2)

1. Application of herba Dendrobii polysaccharide in preparing medicine for treating retinoblastoma is provided.

2. The use of claim 1, wherein the dendrobium nobile polysaccharide is prepared by the following steps:

step one, crushing: taking fresh dendrobium stem strips, removing leaves, cleaning and crushing for later use;

step two, adding 2-6 times of water into the fragments obtained in the step one according to the mass ratio, soaking for a period of time, filtering to obtain filter residues and filtrate, and repeating the soaking and filtering operation on the filter residues for 1-4 times to obtain 2-5 times of filtrate; step three, combining the filtrates, carrying out vacuum rotary evaporation and concentration to obtain a concentrated solution, centrifuging, and keeping a supernatant;

step four, adding 3-5 times of volume of absolute ethyl alcohol into the concentrated solution obtained in the step three to carry out alcohol precipitation;

and fifthly, centrifuging the solution precipitated in the fourth step, removing the supernatant, dissolving the precipitate with water, then carrying out vacuum rotary evaporation concentration, and carrying out freeze drying to obtain the dendrobium nobile polysaccharide.

Images