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New use of tripchlorolide,T4

A technology of chloractone and tripterygium, which is applied in the new field of application of tripterygium chloracterol T4, can solve the problems of high toxicity of crude extracts and limited clinical application, achieve high development and application value, and reduce the number of neurons The damage, the effect of protecting survival

Inactive Publication Date: 2008-12-31
FUJIAN MEDICAL UNIV UNION HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In summary, a large number of basic research and many clinical studies have shown that the extract of Tripterygium wilfordii has obvious anti-inflammatory and immunoregulatory effects on the process of neuroimmune inflammation, but the toxicity of the crude extract of Tripterygium wilfordii limits its use. Clinical application

Method used

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  • New use of tripchlorolide,T4
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  • New use of tripchlorolide,T4

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1. Tripterygium Chlorolactone T 4 Effects on neuronal and microglial survival

[0030] method:

[0031] MTT reduction method detection

[0032] (1) Neruo-2A cells, BV-2 cells, and primary microglia were inoculated in a 96-well culture plate, and 100 μl (1×10 4 cells / well), the cells can be used for drug intervention after attachment; the primary mouse cortical neuron cells are 100 μl per well (initial inoculation 1×10 5 cells / well), on the 6th day of in vitro culture for drug intervention.

[0033] (2) Different concentrations (0, 5, 10, 20, 40, 80nM) of triptolide T 4 After acting for 48 hours, 10 μl of 5 mg / ml MTT solution was added to each well, and incubated at 37° C. for 4 hours.

[0034] (3) Add stop solution (containing 10% SDS, 5% isopropanol, 0.012N hydrochloric acid) to terminate the culture, and incubate overnight at 37°C.

[0035] (4) The next day, measure the OD570 value of each well with a 570nm wavelength with a microplate instrument, and...

Embodiment 2

[0040] Embodiment 2. Tripterygium chloractone T 4 Alleviate oligomeric Aβ 1-42 / LPS-induced glial-inflammatory response to neuronal toxic damage

[0041] method:

[0042] 1. MTT reduction method detection: the specific operation method is the same as the MTT reduction method detection described in Example 1.

[0043] 2. Immunofluorescence Cytochemistry

[0044] (1) Primary cortical neurons cultured in vitro on day 6 were fixed with 4% paraformaldehyde at room temperature for 30 min;

[0045] (2) Remove the fixative, wash with PBS three times, each time for 3 minutes;

[0046] (3) Block with TBSTx containing 5% BSA for 60 min, and shake gently on a shaker;

[0047] (4) Remove the blocking solution, incubate with anti-MAP-2 antibody (1:1000) diluted in TBSTx containing 5% BSA, overnight at 4°C;

[0048] (5) Remove the primary antibody diluent, wash with TBSTx three times, each time for 5 minutes, and shake gently on the shaker;

[0049] (6) Remove the washing solution, ad...

Embodiment 3

[0062] Example 3. Tripterygium Chlorolactone T 4 oligomeric Aβ 1-42 Activated microglia produced effects on the levels of inflammatory mediators (TNF-α, IL-1β, PGE2 and NO).

[0063] method:

[0064] 1. Cell culture supernatant TNF-α, IL-1β and PGE 2 Determination of products

[0065] triptolide T 4 Treat for 60min and then use Aβ 1-42 Stimulate for 24h to detect TNF-α, PGE 2 , or stimulated for 3h for the detection of IL-1β. The supernatant was collected after centrifugation and stored at -80°C if not detected immediately. Mouse TNF-α, IL-1β were detected by ELISA kit (purchased from BioSource Company), supernatant PGE 2 Detected by EIA kit (purchased from Cayman Company). The operating procedure was according to the kit instructions.

[0066] 2. Determination of NO metabolite nitrite

[0067] Triptolide T 4 Treat for 60min and then use Aβ 1-42 After being stimulated for 24 hours, the supernatant aspirated after centrifugation was used to detect the extracellular...

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Abstract

The present invention relates to the usage of wilfordii chlorine lactonic alcohol T4 for curing neural immune inflammatory diseases and provides a new usage of the wilfordii chlorine lactonic alcohol T4 for curing neural immune inflammatory diseases caused by the neural toxicity of microglia-mediated ABeta1-42 and LPS. The traditional medicine monomer of the wilfordii chlorine lactonic alcohol T4 has the following functions: obviously reducing the toxic damages of the oligmeric ABeta1-42 and LPS-induced gel-inflammation reaction on nerve cells, so as to protect the survival of nerve cells; inhibiting the level of inflammation medium produced by the oligmeric ABeta1-42 and the LPS-induced microglia cells; inhibiting the increase of iNOS and COX-2 expression of the oligmeric ABeta1-42 and the LPS-induced microglia cells; inhibiting the activation of NF-KappaB and JNK signal access of the oligmeric ABeta1-42 and the LPS-induced microglia cells. Therefore, the present invention provides direct powerful experimental evidence and theoretical basis for the application of the wilfordii chlorine lactonic alcohol T4 in neural immune inflammatory diseases, especially prevention and treatment of Alzheimer disease, thus having strong scientificity and creativity, as well as high development and application value.

Description

technical field [0001] The present invention relates to tripterygium chloractone T 4 Use for the treatment / prevention of neuroimmune inflammatory diseases, in particular tripterylide chloractone T 4 In the treatment / prevention of endotoxin lipopolysaccharide and Aβ 1-42 New uses for neuroimmune-inflammatory diseases caused by mediated neurotoxic effects. Background technique [0002] Alzheimer's disease (AD) is a common neurodegenerative disease with memory and cognitive dysfunction as the main clinical features, which seriously endangers the physical and mental health of middle-aged and elderly people. The formation of neuroinflammatory plaques (Neuriticplagues, NPs) and neurofibrillary tangles (Neurofibrillary tangles, NFTs) in neurons, as well as the loss of neurons and synapses. Although the etiology and pathogenesis of AD have not yet been fully elucidated, it is generally believed that β-amyloid (amyloid-beta, Aβ) is the main pathogenic factor of AD, so the "Aβ casc...

Claims

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Application Information

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IPC IPC(8): A61K31/585A61P25/28A61P25/00A61P29/00
Inventor 陈晓春
Owner FUJIAN MEDICAL UNIV UNION HOSPITAL
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