Chinese Herbal Aqueous Extract Having Anti-Anxiety Activities and Method of In Vitro Evaluating the Same

a technology of anti-anxiety activity which is applied in the field of active substances having anti-anxiety activities and in vitro evaluation of the same, can solve the problems of anxiety that cannot be effectively improved, existing anti-anxiety drugs or sleeping drugs, and the nerve system suffers injury under oxidative pressure and hyper-excitatory electrical potential, and achieves the effect of enhancing the expression of the cannabinoid receptor type-1 (cb1) in the membran

Inactive Publication Date: 2012-08-30
NAT PINGTUNG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0011]Moreover, a Chinese herbal aqueous extract with anti-anxiety activity is provided. The Chinese herbal aqueous extract, which is evaluated by using the aforementioned method, can enhance a membrane expression of a Cannabinoid receptor type-1 (CB1).
[0017]According to an embodiment of the invention, an expression amount of a Cannabinoid receptor type-1 (CB1) is enhanced in vivo by the aforementioned Chinese herbal aqueous extract with anti-anxiety activity.

Problems solved by technology

However, the nervous system suffers injury under oxidative pressure and hyper-excitatory electrical potential.
Such anxiety cannot be effectively improved even through the administration of anti-anxiety drugs.
However, the existing chemical drugs of anti-anxiety drugs or sleeping drugs have some disadvantages, such as temporary and permanent amnesia in long-term usage due to high effect concentration and most stimulation of the inhibitory potential of such drugs.
Moreover, there is no effective, quick and economic in vitro evaluation method for fast screening natural anti-anxiety active constituents from numerous raw materials of Chinese herbal medicines.

Method used

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  • Chinese Herbal Aqueous Extract Having Anti-Anxiety Activities and Method of In Vitro Evaluating the Same
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  • Chinese Herbal Aqueous Extract Having Anti-Anxiety Activities and Method of In Vitro Evaluating the Same

Examples

Experimental program
Comparison scheme
Effect test

example 1

Establishment of In Vitro Cell Platform

[0046]1. Cell Culture of Human Neuroblastoma Cell Line IMR-32 and Human Glioblastoma U-87 MG

[0047]Human neuroblastoma cell line IMR-32 (deposited at ATCC under accession No.: ATCC CCL-127) and human glioblastoma U-87 MG (deposited at ATCC under accession No.: ATCC HTB-14) were purchased from the Food Industry Research and Development Institute (FIRDI), Bioresource Collection and Research Center (BCRC), Hsinchu, Taiwan). The cell lines IMR-32 and U-87 MG were cultured in the minimum essential medium (MEM; Gibco® BRL, Grand Island, N.Y.) that contained 2 mM L-glutamine, 1 mM non-essential amino acids, 10% fetal bovine serum (FBS), 0.1 mg / mL streptomycin and 0.5 mg / mL ampicillin. Those cells were incubated for 48 hours in an incubator (NUAIR NU4500, USA) with 5% CO2 humidified atmosphere at 37° C., and changed with the fresh medium every two or three days. When those cells reached about 90% confluence, the cells were gently washed in phosphate buf...

example 2

In Vitro Evaluating Chinese Herbal Aqueous Extract

[0055]This EXAMEPLE was directed to evaluate the anti-anxiety activities of the Chinese herbal aqueous extracts of EXAMPLE 1 in vitro by using the epileptic cell platform established by EXAMPLE 1.

[0056]1. Evaluation of Cell Viability

[0057]In this EXAMPLE, cell viability of EXAMPLE 1 was detected by using MTT test. At first, methylthiazoletetrazolium (MTT; Sigma, St. Louis, Mo., USA) solution was prepared by adding 5 mg MTT into 1 mL PBS. Next, each well (i.e. 1×105 cells / well) of 96-well microplate was added with 10 μL MTT solution, incubated for 24 hours in the incubator (NUAIR NU4500, USA) in the dark with 5% CO2 humidified atmosphere at 37° C., and then centrifuged for 10 minutes in a rotation speed of 1,000×g at 4° C. The supernatant was discarded and 100 μL dimethyl sulfoxide (DMSO; Merck, Darmstadt, Germany) solution is added. The microplate was vibrated gently for 10 minutes. Absorbance at 550 nm (OD550 nm) of each well was de...

example 3

In Vivo Evaluating Anti-Anxiety Activities of Chinese Herbal Aqueous Extract by Using Experimental Animal Model

[0070]This EXAMEPLE was directed to evaluate the anti-anxiety activities of the Chinese herbal aqueous extracts of EXAMPLE 1 in vivo by using an animal model.

[0071]1. Establishment of Experimental Animals

[0072]1.1 Care of Experimental Animals

[0073]In this EXAMPLE, fifty six-week-old specific pathogen-free (SPF) imprinting-control-region (ICR) mice (purchased from BioLASCO Taiwan Co., Ltd., Yi-Lan, Taiwan) were randomly allocated into four experimental groups (i.e. PO group, SBO group, AC group and SZS group) and one control (PBS) group, each group of which had ten mice.

[0074]All ICR mice were bred in a SPF environment with positive air pressure. Ambient temperature was controlled at 24° C., relative humidity at 65±5. In addition, the animals were maintained on a reverse 12 h light-dark cycle. Mice were provided with standard laboratory chow and water ad libitum. All experim...

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Abstract

A Chinese herbal aqueous extract having anti-anxiety activities and a method of in vitro evaluating the same are disclosed. Neural cells and / or glial cells are cultured in a medium containing the Chinese herbal aqueous extract, and then subjected to an electric pulse treatment to form epileptic cells. The epileptic cells are applied on evaluation of at least one criterion of anti-anxiety activities of the Chinese herbal aqueous extract.

Description

RELATED APPLICATIONS[0001]This application claims priority to Taiwan Application Serial Number 100106448, filed February 2011, which is herein incorporated by reference.BACKGROUND[0002]1. Field of Invention[0003]The present invention relates to an active substance having anti-anxiety activities and a method of evaluating the same. More particularly, the present invention relates to a Chinese herbal aqueous extract and a method of in vitro evaluating anti-anxiety activities of the same.[0004]2. Description of Related Art[0005]Animal central nervous system (CNS) has many kinds of receptor systems for receiving and responding signals that generate from body under environmental changes. Some active natural components, which can keep spirit and mind health, perhaps function on many CNS receptors including cannabinoid receptor 1 (CB1) and other receptors. However, the nervous system suffers injury under oxidative pressure and hyper-excitatory electrical potential. Therefore, reduction of ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K36/725A61P25/22G01N33/53A61K36/13C12Q1/02A61K36/254
CPCA23L1/3002G01N2800/301G01N2510/00G01N33/5058A23L33/105A61P25/22A61K36/254A61K36/725G01N33/502G01N2333/415G01N2500/10
Inventor CHAUNG, HSO-CHIYANG, TSUNG-HUICHEN, HOLICHEN, CHIH-CHENGCHIU, HSIEN-HSUEH
Owner NAT PINGTUNG UNIV OF SCI & TECH
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