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Homogeneous snakegourd root polysaccharide with immunization activity and antitumor effect, and application thereof

A technology of trichosanthin polysaccharide and anti-tumor effect, applied in the field of medicine and biology, can solve the problem of little research on polysaccharide components, and achieve the effect of good immune activity and good anti-tumor effect

Inactive Publication Date: 2015-08-12
JIAXING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are few studies on the polysaccharide components in trichosanthes

Method used

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  • Homogeneous snakegourd root polysaccharide with immunization activity and antitumor effect, and application thereof
  • Homogeneous snakegourd root polysaccharide with immunization activity and antitumor effect, and application thereof
  • Homogeneous snakegourd root polysaccharide with immunization activity and antitumor effect, and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1: Preparation of Trichosanthus trichosanthis total polysaccharide

[0017] Grind the medicinal material Trichosanthes kirilowii Maxim (dried root of Trichosanthes kirilowii Maxim, place of origin: Pinghu, Zhejiang) with a pulverizer, take 500g of the powder, reflux and degrease with petroleum ether for 8 hours, add 15 times the amount of water to the dregs, extract at 70°C for 2 hours, repeat 3 times, Centrifuge, combine the supernatants, concentrate under reduced pressure to 1 / 4 of the original volume, add 95% ethanol to make the final alcohol concentration 80%, stand still at room temperature for 10 hours, and precipitate the crude polysaccharide precipitate. Use trichloroacetic acid (TCA) method to deproteinize, make the TCA concentration in the solution reach 6%, place, centrifuge to remove protein precipitate, there is no absorption peak of protein and nucleic acid by ultraviolet scanning 260, 280nm. The supernatant was dialyzed against distilled water ...

Embodiment 2

[0018] Embodiment 2: the preparation of homogeneous trepan pollen polysaccharide (RTPS-I)

[0019] Get the trichosanthes total polysaccharide that embodiment 1 obtains through DEAE-cellulose (OH - , 4cm×60cm) column chromatography, with 0 ~ 0.5mol. L -1 Gradient elution with NaCl, one tube was collected every 10 minutes, 6 mL per tube. Sulfuric acid-phenol method was used for color development, the sugar-containing fraction was collected, dialyzed, concentrated, and dried, and the polysaccharide fraction was put on DEAE-Sepharose FF (Cl - , 2.5cm×50cm) chromatographic column, 0~0.8mol﹒ L -1 NaCl (pH7.5) gradient elution, sulfuric acid-phenol method for color development, the eluted fractions were collected, dialyzed, concentrated, and freeze-dried. Among them, the trichosanthin polysaccharide RTPS-I was the most abundant. The above polysaccharide fraction RTPS-I was then treated with Sephadex G-200 (Cl - , 2.6cm×90cm) gel filtration chromatography for further purificatio...

Embodiment 3

[0020] Embodiment 3: The immune activity test of uniform day pollen polysaccharide (RTPS-I)

[0021] (1) All day pollen polysaccharide (RTPS-I) promotes the proliferation of human PBMC lymphocytes.

[0022] Human peripheral blood mononuclear cells (PBMCs) were seeded in 96-well culture plates at 100 μL per well, and incubated at 37°C, 5% CO 2 After 24 hours of pre-cultivation in the incubator, the homogeneous 10mmol. L -1 Phytohemagglutinin (PHA) was used as the positive control group, and the same amount of 10% FCS RPMI 1640 was used as the negative control group. After 37°C, 5% CO 2 After cultivating for 72 hours, 2000r. min -1 Centrifuge for 10min, carefully absorb the supernatant, and add 5mg﹠ to each well. mL -1 Tetrazolium blue (MTT) 50 μ L, continue to incubate for 4 hours. After the supernatant was discarded by centrifugation, 150 μL of dimethyl sulfoxide (DMSO) was added to each well, shaken by an oscillator for 20 min, and the absorbance value was detected at...

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Abstract

The invention provides a homogeneous snakegourd root polysaccharide with immunization activity and antitumor effect, and an application thereof. The polysaccharide is separated and extracted from a traditional Chinese medicine snakegourd root, is a white flocculent solid, can be easily dissolved in water, and is composed of C, H and O, the specific rotation [alpha]<D><50>(H2O) is +165.58, the weight-average molecular weight (Mw) is 1.76*10<4>, and the main glycosyl group of the polysaccharide is glucose. RTPS-I can promote proliferation of human peripheral blood mononuclear cells (PBMCs), increases T lymphocyte subgroups in the PBMCs, and stimulates the human PBMCs to secrete TNF-alpha and IL-6 cytokines, and the immune activity of the RTPS-I is obviously better than that of snakegourd root total polysaccharides; and the RTPS-I can substantially inhibit the growth of human cervical carcinoma cells (HeLa cells) and human breast cancer cells (MCF-7 cells). The homogeneous snakegourd root polysaccharide can be used for exploiting immunoloregulation adjuvants, and can also be used as an antitumor drug lead compound.

Description

technical field [0001] The invention relates to the polysaccharide of the bioactive component of the traditional Chinese medicinal material Trichosanthemi chinensis, and belongs to the field of medical biotechnology. Background technique [0002] As a traditional Chinese herbal medicine, Radix Trichosanthis has more than a thousand years of experience in clinical application of traditional Chinese medicine. Clinically, it mainly treats ectopic pregnancy, hydatidiform mole, residual placenta and trophoblastic tumor. Modern studies have shown that trichosanthum is rich in various chemical components such as protein, polysaccharide, lectin, saponin and flavonoids. Among them, trichosanthin has been isolated and identified, and it has been confirmed that it has various biological activities such as anti-tumor. There are few studies on the polysaccharide components in trichosanthes. Contents of the invention [0003] The first object of the present invention is to provide a ...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61P37/02A61P35/00
Inventor 屠婕红徐水凌黄佳
Owner JIAXING UNIV
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