Method for enhancing heat generation efficiency of brown adipocytes and application

A technology of brown fat and cells, which is applied in the field of enhancing the heat production efficiency of brown fat cells, and can solve the problems of low heat production efficiency

Pending Publication Date: 2018-01-30
上海聿健生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, at present, drug research using BAT thermogenesis to treat obesity has encountered obstacles (Carey et al., 2013; Larsen et al.,

Method used

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  • Method for enhancing heat generation efficiency of brown adipocytes and application
  • Method for enhancing heat generation efficiency of brown adipocytes and application
  • Method for enhancing heat generation efficiency of brown adipocytes and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0239] Example 1 NE-induced thermogenic response of brown adipocytes

[0240] The mitochondrial temperature detection method based on rhodamine B methyl ester (RhB-ME) and rhodamine 800 (Rh800) is used to detect the heat production in cells (for specific methods, please refer to: New temperature-sensitive fluorescent compounds and their applications, patent application number 201410850735.3 ). In short, there are a large number of mitochondria in brown adipocytes, which are the main places for generating heat in cells. Using the mitochondrial temperature measurement method to study the heat production of brown adipocytes, a large number of mitochondrial heat production can be obtained at the single-cell level. Data, thus reflecting the thermogenesis of brown adipocytes. Fluorescent dyes RhB-ME and Rh800 can be located in the mitochondria of brown fat cells. The fluorescence intensity of RhB-ME changes with temperature, while the fluorescence intensity of Rh800 is basically co...

Embodiment 2

[0243] Example 2 NE activates the ATP-dependent proton pump (H + -ATPase)

[0244] The primary cultured brown adipocytes after electroporation expressed the ATP-sensitive fluorescent protein, placed in Tyrode solution, stimulated with norepinephrine, and the FRET signal of the ATP-sensitive fluorescent protein was observed and imaged in real time by confocal laser. Graphical data analysis is then performed.

[0245] To test whether NE activates the ATP-dependent proton pump (H + -ATPase), using cytoplasmic and mitochondrial ATP-sensitive fluorescent proteins AT1.03 and mitAT1.03 (Imamura et al., 2009), to detect changes in cytoplasmic and mitochondrial ATP when NE stimulated brown adipocytes ( figure 2 , A-E). It was found that 0.1μM NE stimulated brown adipocytes to cause a decrease in cytoplasmic and mitochondrial ATP, suggesting that NE stimulated activation of a certain ATPase (ATPase). Considering that only mitochondrial complex V of mitochondrial complexes I-V can c...

Embodiment 3

[0246] Example 3 Oligomycin or EGCG combined with NE can significantly improve the heat production efficiency of brown adipocytes

[0247] Primary cultured brown adipocytes stained with rhodamine B methyl ester (RhB-ME) and rhodamine 800 (Rh800) were placed in Tyrode solution, co-stimulated with norepinephrine and mitochondrial complex V function inhibitor, laser Confocal real-time observation and imaging of thermosensitive and mitochondrial membrane voltage signals, followed by graphic data analysis.

[0248] If it is mitochondrial complex V that causes the hypothermia of NE-induced brown adipocytes, the present invention speculates that inhibiting the function of mitochondrial complex V should enhance the heat production efficiency of NE-induced brown adipocytes. Experimental results ( image 3 ) showed that NE could induce thermogenesis in almost all brown adipocytes in the presence of oligomycin to inhibit the function of mitochondrial complex V ( image 3 , A-C), the mi...

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Abstract

The invention provides a method for enhancing the heat generation efficiency of brown adipocytes and application, and concretely provides a purpose of a heat generation enhancing compound for preparing a preparation or a composition. The preparation or the composition is used for enhancing the heat generation efficiency of the brown adipocytes induced by a downstream cAMP-PKA signal channel and anNE (Norepinephrine) receptor, wherein the heat generation enhancing compound is a plastosome compound V/ATP (Adenosine Triphosphate) hydrolase inhibitor. When the heat generation enhancing compound and a first compound (an NE receptor and a downstream cAMP-PKA signal channel enhancer) are combined, the heat generation efficiency of the brown adipocytes induced by the downstream cAMP-PKA signal channel and the NE (Norepinephrine) receptor can be obviously enhanced.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method and application for enhancing the heat production efficiency of brown adipocytes. Background technique [0002] The occurrence of obesity is mainly the depletion of excess nutrients from ingested nutrients (Celi, 2009). Obesity is one of the important factors leading to diseases such as type 2 diabetes, hyperlipidemia and cancer. Existing methods for treating obesity mainly include dieting, strengthening exercise, drug therapy, etc.; dieting and strengthening exercise have certain effects, but it is difficult to persist for a long time, and the effect of drug treatment is very limited (Celi, 2009; Tseng et al., 2010), Moreover, such weight-loss drugs lack a complete evaluation of their effects and side effects, resulting in a greater risk of drug use. [0003] The energy of nutrients is not only used for external work consumption, but also consumed in the form of heat prod...

Claims

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Application Information

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IPC IPC(8): A61K45/06A61K31/4412A61K31/12A61P3/04C12Q1/02
Inventor 康建胜谢涛嵘
Owner 上海聿健生物科技有限公司
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