KR20140120639A - Composition for preventing Obesity or Hyperlipidemia Comprising tert-butylhydroquinone(tBHQ) - Google Patents

Abstract

The present invention relates to a component or composition containing tert-butylhydroquinone (tBHQ) as an active ingredient for losing weight and preventing hyperlipidemia. According to an embodiment of the present invention, the present invention provides the component or composition containing tert-butylhydroquinone (tBHQ) as an active ingredient for losing weight and preventing hyperlipidemia. According to a preferable embodiment of the present invention, the present invention provides a component or composition for losing weight and preventing hyperlipidemia, wherein the component or composition contains 0.00001-0.001% of tert-butylhydroquinone (tBHQ).

Description

TECHNICAL FIELD The present invention relates to a composition for prevention of obesity or hyperlipidemia comprising tetrabutylhydroquinone as an active ingredient (Composition for preventing Obesity or Hyperlipidemia Comprising tert-butylhydroquinone (tBHQ)

The present invention relates to a composition comprising tetrabutylhydroquinone (tBHQ), a component for reducing hyperuricemia, and a component for preventing hyperlipidemia.

Obesity is caused by genetic or specific hormonal disorders, but is caused by excessive calorie intake and lack of physical activity. Thus, obesity generally occurs when the amount of energy consumed is less than the energy consumed. Obesity is a cause of many complications such as insulin resistance, hyperlipidemia, hypertension, atherosclerosis and the like, and the obesity population is increasing all over the world.

Low calorie intake and energy expenditure through exercise are the main treatments for obesity. However, methods that can be used to inhibit obesity by using drugs include ways of inhibiting ingestion of food, especially fat and sugar, and inhibiting fat accumulation , How to promote the burning of accumulated fat, and how to suppress appetite.

Drugs using this method have been developed for the control of obesity. Orlistat is used to inhibit the lipases of the pancreas and to partially inhibit the production of fatty acids and triglycerides (TG). Sibutramine has been used as an appetite suppressant, but has recently been withdrawn from the market because of a reported mental disorder and the risk of myocardial infarction or stroke.

In addition, hyperlipidemia is defined as a serum concentration of one or more of serum lipids such as cholesterol, triglyceride, phospholipid, and free fatty acid. The normal range of serum lipid in fasting is 50 ~ 150 mg / dl of triglyceride, 250 mg / dl, cholesterol 130 ~ 230 mg / dl, and free fatty acid 5 ~ 10 mg / dl. Left untreated hyperlipemia increases the likelihood of causing severe complications such as hypertension, coronary atherosclerosis (angina pectoris, myocardial infarction), and cerebral atherosclerosis (cerebral infarction).

On the other hand, thiazolidinediones, pioglitazone and rosiglitazone, which are peroxisome proliferator receptor gamma (PPAR-γ) agonists, were used as drugs to improve insulin sensitivity. Fibrates reduce 23-80% of TG levels and 38-80% of LDL cholesterol levels with peroxisome proliferator receptor alpha (PPARa) agonists (Srivastava, 2011; Zaborska et al., 2000; Millar et al., 2009 ).

Fibrates can reduce atherosclerotic hyperlipidemia and cardiovascular disease (CVD) (Feitosa et al., 2011; McKeage and Keating, 2011). It has been reported that continuous activation of PPARa by Huang inhibits obesity (Huang et al., 2012). Thus, moderate activation of PPARa, which is not excessive, is likely to be a good drug to ameliorate obesity and metabolic diseases.

Using acyl-CoA oxidase 1 (ACOX1, the first enzyme of the peroxisomal fatty acid β oxidation system) mouse, which is a gene dependent on the activation of PPARa to detect new candidate substances that improve the risk of metabolic syndrome, obesity and cardiovascular disease I created a search model. ACOX1 expression was amplified and ligated to a luciferase enzyme gene, and then introduced into HepG2 cells, a human liver cancer cell, to develop a cell line capable of monitoring the expression of ACOX1.

ACOX1 is the first enzyme involved in lipid oxidation in the β-oxidation system associated with fatty acid metabolism. It has been found that tet-butylhydroquinone (tBHQ) increases the expression of this enzyme. This means that tBHQ increases the expression of ACOX1 gene due to activation of PPARa, and that the increase of ACOX1 expression contributes to the promotion of lipid metabolism and the decrease of fat accumulation.

tBHQ is a food grade antioxidant food additive approved by the FDA, but no report on the effects of improving obesity and hyperlipidemia. The total concentration of tBHQ available as food is allowed to not exceed 200 ppm (0.02%) based on fat and oil content (Gharavi et al., 2007).

tBHQ prevents oxidative stress and apoptosis in cells, and protects acute kidney damage induced by cis-diamminedichloroplatinum II in rats. In contrast, many adverse effects associated with overuse have been reported (Shibuya et al., 2012; Yu et al., 2000; Peters et al., 1996).

Despite the history of tBHQ, which has been used for over 30 years, the effects of weight reduction and hyperlipidemia have not been reported. Thus, the present inventors repeatedly studied the effects of tBHQ on the suppression of weight and the improvement of hyperlipemia, thereby completing the present invention.

The object of the present invention is to provide a composition for the prevention of hyperlipemia and obesity, which contains tBHQ, for the purpose of improving blood lipid levels and preventing obesity, for the prevention of hyperlipemia and obesity.

In order to achieve the above object, the present invention provides a composition for preventing weight loss and hyperlipidemia comprising tetrabutylhydroquinone (tBHQ) as an active ingredient.

According to a preferred embodiment of the present invention, the present invention provides a composition and composition for preventing weight loss and hyperlipemia, wherein the content of tetrabutylhydroquinone (tBHQ) is 0.0001 to 0.001%.

According to the present invention, mice fed a diet containing 0.0002% to 0.001% of tBHQ in the diet were found to have a weight loss effect of 31.8% to 61.4% ( P <0.05), blood glucose, insulin and leptin levels, plasma triglyceride Total cholesterol concentration can be lowered.

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to improve blood lipid levels, prevent obesity and prevent hyperlipidemia and obesity, and to use it as a health food for the prevention of hyperlipidemia and obesity. Further, according to the present invention, it is characterized by reducing the size of adipocytes.

FIG. 1 is a graph showing changes in body weight after ingestion of a general formula, high fat diet (HFD), HFD + BHQ (0.0002%) and HFD + BHQ (0.001%), respectively.
FIG. 2 shows a composition and content table of a control group and a high fat feed (HFD).
FIG. 3 shows the expression of ACOX1-promoter-dependent gene expression in HepG2 cells by the activity of luciferase enzyme (a). The efficacy of the previously known drug, fenofibrate, was also compared (b).
FIG. 4 shows the results of measurement of ACOX1 mRNA levels expressed in liver of mice receiving tBHQ. Real-time RT-PCR showed that tBHQ (10 mg / kg, w / w) increased ACOX1 mRNA with time.
Figure 5 shows the effect of tBHQ on expression of luciferase enzyme dependent on pPPRE in hepatocytes. tBHQ is a concentration-dependent increase in the expression of luciferase dependent on pPPRE in HepG2 cells (n = 3-5. * P <0.05, ** P <0.01).
FIG. 6A shows the amounts (a) and (b) intake of C57BL / 6 mice fed high fat diets (HFD) containing the components of the invention for 32 days. 6B is a graph showing the weight of liver (a), epididymal fat (b), mesentric fat (c) and Peri-kidney fat (d) after dietary administration in each example (n = 6, * P < 0.05).
FIG. 7 shows the results of total cholesterol, triglyceride, HDL cholesterol, blood glucose, insulin, and leptin levels measured after 32 days of dieting (n = 6. * P <0.05, ** P < ).
Figure 8 is a graph showing the effect of tBHQ on fat accumulation in liver tissue (a) and adipocyte tissue (b) in mice fed a 32-day diet (n = 6. * P < 0.05) .

The present invention relates to a composition for preventing obesity or hyperlipidemia comprising tetrabutylhydroquinone as an active ingredient, and the present invention will be described in detail with reference to the following examples.

[ Example  One : ACOX1 On Expression of &lt; RTI ID = 0.0 &gt;

3,780 nucleotides (-1 to -3780 sites) of the promoter region of the pACOX1 gene of mice were amplified with primers mACOX1-F (5'-TATTTGGGATCCAAGCTTCTGCTA-3 ') and mACOX1-R (5'-AATTCGAATTCATGACGGAGACCATGGCT-3'). The amplified DNA was bound to BamHI and HindIII sites and introduced into HepG2 cells to produce a permanent cell line. The efficacy of tBHQ was assessed using this cell line and fenofibrate was used as a positive control (see FIG. 3).

[ Example  2 : tBHQ Transferred by ACOX1 Check the amount]

Twenty-four C57BL / 6 mice were divided into four groups and IP injection of tBHQ (10 mg / kg) to determine the amount of ACOX1 transduced by tBHQ. ACOX1 mRNA was quantitated with real-time RT-PCR instrument. cDNA was amplified using two kinds of primers 5'-CAGCCAGATTGGTAG AAATTGCT-3 'and 5'-ACGCCACTTCCTTGCTCTTC-3', GAPDH was amplified using 5'-TGGAAGAGTGGGAGTTGCTGTTG-3 'and 5'-CGTGCCGCCTGG AGAAACC-3' The amount of transcribed ACOX1 gene was corrected. The reaction time was measured in real time at 95 ° C for 20 seconds, 55 ° C for 10 seconds, and 72 ° C for 30 seconds. The relative values of ACOXl mRNA were quantified by CT values (see FIG. 4) .

 [ Example  3: tBHQ &Lt; / RTI &gt;

The pPPRE-Luc2 vector was constructed by ligating three times the DNA of 5'-TGACCTTTGTCCTAGGACA-3 'DNA to the XhoI and HindIII sites of the pGL4.20 plasmid and putting it into human hepatoma cell HepG2 to form a permanent cell line The efficacy of tBHQ was assessed (see FIG. 5) .

[ Example  4: Mouse blood sample collection and analysis]

Uninfected 9 week old male C57BL / 6 mice from certain pathogens were purchased from Samtako (Osan, Korea) and allowed to adapt to the environment for one week before use. The mice were allowed free access to food and water and the environment was maintained at a temperature of 20 ± 1, humidity of 55% ± 5% and a 12-hour day and night cycle.

Twenty-four mice were randomly divided into four groups and divided into two groups: high fat diets (D12492, New Jersey, USA) and high fat diets (0.0002%, 0.001% tBHQ) Respectively. Weight and dietary intake were measured twice a week. At the end of the experiment, levels of leptin, insulin and blood glucose were analyzed through blood samples. Liver fat accumulation, epididymal fat, mesenteric fat and kidney fat were measured (see FIG. 6) .

[ Example  5: Biochemical analysis of plasma]

The levels of total cholesterol, triglyceride, HDL cholesterol, and blood glucose were measured using a chemical analyzer (AU400, Olympus, Tokyo, Japan). Plasma leptin and insulin levels were measured using an ELISA assay kit (for mice) (see FIG. 7) .

[ Example  6: Dyeing and measurement]

The hepatocyte was 7-μm thick and stained with hematoxylin and eosin. The adipose tissue was determined by staining 7 ㎛ thick epididymal fat slices with hematoxylin and eosin to separate the cell matrix and nucleus (see FIG. 8) .

(HFD), HFD + BHQ (0.0002%) and HFD + BHQ (0.001%), respectively. The composition and content table of the composition are shown in Fig.

BHQ + HHD + BHQ (0.001%) was higher than BHQ + HHD + BHQ (0.0002), while BHQ + HFD + BHQ was higher than that of the high fat diet %), The increase rate of body weight was found to be low.

3 (a) shows the results of measuring the expression level of ACOX1-promoter-dependent gene expression in HepG2 cells as an activity of luciferase enzyme, and Fig. 3 (b) This is also the result of comparing the efficacy of fenofibrate. tBHQ is almost similar to the ACOX1 activity increasing effect of the conventional drug fenofibrate.

FIG. 4 shows the results of real-time RT-PCR of tBHQ (10 mg / kg, w / w) as a result of measurement of ACOX1 mRNA level in liver of mice in which tBHQ was administered I could.

Figure 5 shows the effect of tBHQ on expression of luciferase enzyme dependent on pPPRE in hepatocyte, indicating that tBHQ increases concentration-dependent expression of luciferase dependent on pPPRE in HepG2 cells (n = 3 to 5. * P <0.05, ** P <0.01).

FIG. 6A shows the amounts (a) and (b) intake of C57BL / 6 mice fed high fat diets (HFD) containing the components of the invention for 32 days. 6B is a graph showing the weights of liver (a), epididymal fat (b), mesentric fat (c) and Peri-kidney fat (d) after dietary administration (n = 6, * P < 0.05), the increase in the weight of liver (a), epididymal fat (b), mesentric fat (c) and Peri-kidney fat (d) was significantly decreased with increasing tBHQ Can be confirmed.

FIG. 7 shows the results of measurement of total cholesterol, triglyceride, HDL cholesterol, blood glucose, insulin, and leptin levels (n = 6. * P <0.05, ** P <0.01 ), The amount of cholesterol, triglyceride, HDL cholesterol, blood glucose, insulin, and leptin was effectively reduced with increasing tBHQ when HFD was taken with tBHQ.

FIG. 8 shows the effect of tBHQ on fat accumulation in liver tissue (a) and adipocyte (b) of mice subjected to 32-day dieting (n = 6. * P < 0.05) When HFD was supplemented with tBHQ, the cell size decreased and the cell number increased in liver tissue and adipocytes.

According to the present invention, mice fed a diet containing 0.0002% to 0.001% of tBHQ in the diet were found to have a weight loss effect of 31.8% to 61.4% ( P <0.05), blood glucose, insulin and leptin levels, plasma triglyceride The total cholesterol concentration was decreased and the fat cell size was decreased.

Therefore, according to the present invention, it is possible to improve blood lipid levels, prevent obesity and prevent hyperlipidemia and obesity, and to use it as a health food for the prevention of hyperlipidemia and obesity.

The above-described embodiments are illustrative and those skilled in the art will be able to make various modifications and alterations to the disclosed embodiments without departing from the spirit of the present invention. The scope of the present invention is not limited by these variations and modifications.

references

1. Feitosa, MF, An, P., Ordovas, JM, Ketkar, S., Hopkins, PN, Straka, RJ, Arnett, DK, and Borecki, IB, Association of gene variants with lipid levels in response to fenofibrate is influenced by metabolic syndrome status. Atherosclerosis , 215, 435-439 (2011).

2. Gharavi, N., Haggarty, S., and El-Kadi, AO, Chemoprotective and carcinogenic effects of tert-butylhydroquinone and its metabolites. Curr . Drug Metab . , 8, 1-7 (2007).

3. Huang, J., Jia, Y., Fu, T., Viswakarma, N., Bai, L., Rao, MS, Zhu, Y., Borensztajn, J., and Reddy, JK, Sustained activation of PPARaby endogenous ligands increases hepatic fatty acid oxidation and prevents obesity in ob / ob mice. FASEB J. , 26, 628-638 (2012).

4. McKeage, K., and Keating, GM, Fenofibrate: a review of its use in dyslipidaemia. Drugs , 71, 1917-1946 (2011).

5. Millar, JS, Duffy, D., Gadi, R., Bloedon, LT, Dunbar, RL, Wolfe, ML, Movva, R., Shah, A., Fuki, IV, McCoy, M., Harris, CJ , Wang, MD, Howey, DC, and Rader, DJ, Potent and selective PPAR-alpha agonist LY518674 upregulates both ApoA-I production and catabolism in human subjects with the metabolic syndrome. Arterioscler . Thromb . Vasc . Biol . , 29, 140-146 (2009).

6. Peters, MM, Rivera, MI, Jones, TW, Monks, TJ, Lau, SS, Glutathione conjugates of tert-butyl-hydroquinone, a metabolite of the urinary tract tumor promoter 3-tert-butyl-hydroxyanisole, are toxic to kidney and bladder. Cancer Res, 56,1006-1011 (1996).

7. Shibuya, N., Kobayashi, S., Yoshikawa, Y., Watanabe, K., Orino, K., Effects of oxidative stress caused by tert-butylhydroquinone on cytotoxicity in MDCK cells. Vet Med Sci, 74, 583-589 (2012).

8. Srivastava, RA, Evaluation of anti-atherosclerotic activities of PPAR-a, PPAR-, and LXR agonists in hyperlipidemic atherosclerosis-susceptible F (1) B hamsters. Atherosclerosis , 214, 86-93 (2011).

9. Yu, R., Mandlekar, S., Kong, A.N., Molecular mechanisms of butylated hydroxylanisole-induced toxicity: induction of apoptosis through direct release of cytochrome c. Mol Pharmacol., 58, 431-437 (2000).

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Claims (2)

A composition for the prevention of weight loss and hyperlipidemia comprising tetrabutylhydroquinone (tBHQ) as an active ingredient. The composition for the prevention of weight loss and hyperlipidemia according to claim 1, wherein the tetrabutylhydroquinone (tBHQ) is contained in an amount of 0.0001 to 0.001%.

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