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Application of oldenlandia diffusa acting on retinol X receptor

A technology of Hedyotis diffusa and retinol, which is applied in the application field of Hedyotis diffusa acting on retinol X receptors, and can solve problems such as unclear anti-tumor mechanism

Inactive Publication Date: 2012-05-02
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Hedyotis diffusa has been proved to have obvious killing effect on various tumors in vitro, animal and clinical experiments, but its anti-tumor mechanism is still unclear

Method used

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  • Application of oldenlandia diffusa acting on retinol X receptor
  • Application of oldenlandia diffusa acting on retinol X receptor
  • Application of oldenlandia diffusa acting on retinol X receptor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 , Preparation of BHP component of Hedyotis diffusa extract

[0026] Take the whole herb of Hedyotis diffusa (purchased from Wujiang Shanghai Caitongde Chinese Medicine Decoction Pieces Company), cut into pieces, soak in 95% ethanol, and reflux for 3 times for 2 hours each time, discard the dregs, combine and concentrate the filtrate, Obtain the crude extract of Hedyotis diffusa.

[0027] Suspend the crude extract of Hedyotis diffusa with water, extract 4 times with petroleum ether until the residual liquid is colorless, and obtain the extract, which is the BHP component (ie, the petroleum ether component of Hedyotis diffusa).

Embodiment 2

[0028] Example 2 : Combination of BHP fraction of Hedyotis diffusa extract and RXR

[0029] Get the Hedyotis diffusa extract BHP component obtained in Example 1, and detect its combination with RXR, the specific process is as follows:

[0030] Add to TEG buffer 3 H 9cis-RA, GST-RXR LBD, and different amounts of Hedyotis diffusa extract BHP components, so that the final reaction system 3 The concentration of H 9cis-RA was 1% (V / V), the concentration of GST-RXR LBD was 0.02g / ml, and the concentrations of BHP components of Hedyotis diffusa extract were 0.1, 1, 10, 100, 1000 μg / mL, Among them, BSA was used instead of GST-RXR LBD as a blank control.

[0031] Incubate the obtained reaction system at 4°C for 16 hours, add GST agarose gel beads, shake at room temperature for 1 hour, centrifuge at 1000 rpm for 1 minute, discard the supernatant, add TEG buffer to wash 3 times, centrifuge, and use Binding buffer to resuspend , add scintillation liquid, measure the cpm value with liq...

Embodiment 3

[0033] Example 3 : BHP fraction of Hedyotis diffusa extract inhibits the transcription of RXR

[0034] CV1 cells (purchased from ATCC) cultured in 10% DMEM (purchased from Hyclone) were divided into 2×10 4 Each well was placed in a 48-well plate, and after 24 hours, each well was co-transfected with the following plasmids with 25 μL Lipofectamine2000 liposomes:

[0035] 10 ng GFP-RXRa, 50 ng TREpal, 50 ng β-Gal, 390 ng pBluescript (purchased from ATCC).

[0036] 6 hours after transfection, use new DMEM medium to replace the medium. After the cells grow for 8 hours, add BHP components to treat for 24 hours. Discard the culture medium, wash with PBS buffer three times, and add 40ul Reporter Lysis Buffer to each well. Cells were lysed and freeze-thawed 3 times.

[0037] Then, detect β-Gal and CAT (chloramphenicol acetyltransferase) activity, the result is as follows figure 2 shown.

[0038] figure 2 The results showed that with the increase of the drug concentration, the...

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Abstract

The invention provides an oldenlandia diffusa extract BHP component used for preparing a drug for regulating the activity and the function of a retinol X receptor. The retinol X receptor is used for selecting an oldenlandia diffusa extract and active ingredients contained therein and used as a receptor binding site for guiding the application of structural modification of the active ingredients contained in oldenlandia diffusa. The oldenlandia diffusa extract BHP component contains ingredients combined with the retinol X receptor. While inhibiting the transcriptional activation of the retinolX receptor, the ingredients can remarkably inhibit the growth of a plurality of cancer cells, wherein the two inhibitions are closely related to each other. As the receptor binding site, the retinol Xreceptor can be used for selecting the active ingredients of the oldenlandia diffusa extract BHP component, obtains a compound drug for regulating the activity of RXR Alpha from the active ingredients, and can be used for guiding the application of the structural modification of the oldenlandia diffusa extract BHP component so as to obtain a compound with higher activity.

Description

technical field [0001] The invention belongs to the field of Chinese herbal medicines, and in particular relates to the application of Hedyotis diffusa acting on retinol X receptors. Background technique [0002] Members of the nuclear hormone receptor family include steroid hormone receptors, thyroid hormone receptors, vitamin D receptors, various orphan receptors, and receptors activated by metabolic intermediates such as fatty acid-activated peroxisome proliferators receptor (PPAR), liver X receptor (LXR) activated by cholesterol metabolites, farnesoid X receptor (FXR) activated by bile acids, and pregnane X receptor activated by exogenous organisms. As a transcription factor, nuclear receptors play an important role in embryonic development, cell differentiation, metabolism and maintenance of body homeostasis. The abnormality of its signaling pathway can lead to proliferation, regeneration and metabolic diseases such as cancer, infertility, obesity and diabetes, etc. . ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K36/74
Inventor 张晓坤任春妍曾锦章
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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