Medicine for inducing cells to die, and its application

A cell and drug technology, applied in the field of biomedicine, can solve problems such as unclear function and mechanism of action, and achieve a significant effect of drug efficacy

Inactive Publication Date: 2011-09-21
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, its exact function and m

Method used

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  • Medicine for inducing cells to die, and its application
  • Medicine for inducing cells to die, and its application
  • Medicine for inducing cells to die, and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1 Astragalus Induces HEL Cell Apoptosis Experiment

[0065] Cell culture and induction of apoptosis: HEL and K562 cells (purchased from the Cell Bank of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) were used in RPMI-1640 medium (GIBCO-BRL) containing 10% calf serum at 37°C with 5% CO 2 cultured in a cell culture incubator. Boil Astragalus (produced in Inner Mongolia) with distilled water for 60-90 minutes, remove lumps, and filter to obtain Astragalus solution. The astragalus solution was centrifuged at low temperature at high speed, and the supernatant was freeze-dried to obtain the astragalus extract, which was stored at -20°C for later use. During induction, add astragalus extract at 4.5 mg / ml to the cultured HEL cells and culture for 3-5 days.

[0066] Flow cytometry analysis: The HEL cells induced and uninduced by Astragalus extract were collected by centrifugation and resuspended in citrate solution. Cell suspensions were fr...

Embodiment 2

[0069] Example 2 Astragalus Induces Up-regulation of Apaf-1 Expression in HEL Cells

[0070] Nucleus staining and morphological observation: HEL cells were collected and fixed, resuspended in Hoechst 33258 solution for staining, and after smearing, observed with a fluorescent microscope and photographed.

[0071] Immunofluorescence staining: The HEL cells induced and uninduced by Astragalus extract were collected by centrifugation, fixed and blocked for 45 minutes, and the primary antibody (Apaf-1 polyclonal antibody was purchased from Santa Cruz Company, sc-8339) was added to react for 30 minutes After washing, Rhodamine-labeled secondary antibody (anti-rabbit, purchased from Rockland Company) was added to react for 30 minutes. Fluorescence microscope observation after washing. After Astragalus induced HEL cells, it promoted the expression of apoptosis protease-activating factor 1 (Apoptotic protease-acting factor 1, APAF1) in the cells and the formation of apoptotic bodies ...

Embodiment 3

[0073] Example 3 Astragalus induced the activity upregulation of caspase-3 in HEL cells

[0074] a) Immunofluorescence staining: The HEL cells induced and uninduced by Astragalus extract were collected by centrifugation, fixed and blocked for 45 minutes, and reacted for 30 minutes with primary antibody (Caspase 3 monoclonal antibody purchased from Santa Cruz Company), washed and then FITC-labeled secondary antibody (anti-mouse, purchased from Rockland Company) was added to react for 30 minutes. Fluorescence microscope observation after washing. After Astragalus induced HEL cells, the activity of protease-3 (Caspase-3) was activated. The results of immunofluorescence analysis showed that after induction of HEL cells with Astragalus, in apoptotic cells, Caspase-3 was activated and stained positively (green, gray in 3a in the attached figure), while in non-apoptotic cells In apoptotic HEL cells, protease-3 activity was negatively stained ( image 3 ).

[0075] The above resul...

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Abstract

A Chinese medicine for treating tumor, especially the tumor in blood system is prepared from the extract of astragalus root and the pharmacologically acceptable carrier and for excipient.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a new application of Astragalus membranaceus and its extract. Astragalus extract can induce apoptosis of HEL cells, indicating that the extract of Astragalus can be prepared as a drug for treating tumors, especially hematological tumors. Background technique [0002] Chinese herbal medicine is a bright pearl in the treasure house of traditional medicine of the motherland. In recent years, it has attracted great attention from western countries. However, little is known about their molecular mechanism of action, which needs to be elucidated using modern knowledge and technology. It is generally believed that astragalus has the effect of enhancing the immune function of the body. Recently, in clinical practice, it is usually used to reduce the toxic and side effects caused by chemotherapy and radiotherapy. However, its exact function and mechanism of action are still unclear. [0003...

Claims

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Application Information

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IPC IPC(8): A61K36/481A61K31/7048A61K9/20A61K9/48A61K9/08A61K9/10A61P35/00
Inventor 程晓东钱若兰侯春晖张学军谢恒月周维影张树冰
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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