• BMB Reports
• /
• v.43 no.4
• /
• pp.257-262
• /
• 2010

The aim of work was to investigate changes of 7-ketocholesterol synthesis in alveolar macrophages in the dynamic of lung mechanical ventilation with injurious parameters. The goal of in vitro part of work was to observe influence of 7-ketocholesterol on iNOS and MIP1 $\beta$ production in bronchoalveolar lavage fluid (BALF) cells. We used 17 healthy domestic pigs randomly assigned into two treatment groups: group I with mechanical ventilation with physiological parameters; group II underwent injurious ventilation with high volume tidal (VT) and low positive end expiratory pressure (PEEP). Cells were analyzed for CYP27A1 protein and gene expression levels, 7-ketocholesterol production. In alveolar macrophages of group II, we obtained increase of production of CYP27A1 protein and 7-ketocholesterol, as well as the expression of the CYP27A1 gene at the 2nd hour of ventilation. In the in vitro experiments we show dose-dependent increase of MIP1 $\beta$ and decrease of CYP27A1, iNOS protein production after 7-ketocholesterol treatment.

• Nutrition Research and Practice
• /
• v.1 no.3
• /
• pp.180-183
• /
• 2007

Cytoprotective ability of polysaccharides isolated from different edible mushrooms was investigated on the 7-ketocholesterol-induced damaged cell line. Polysaccharide extracts from six different edible mushrooms-Flammulina velutipes, Peurotus ostreatus, Lentinus edodes, Agrocybe aegerita, Agaricus blazei, and Cordyceps militaris- were prepared by hot water extraction and alcohol precipitation. Cytoprotective ability was evaluated by measuring the viable cells of the normal embryonic liver cell line (BNL CL. 2) in the presence of 7-ketocholesterol. At $80\;{\mu}g/mL$ of 7-ketocholesterol, cytotoxicity was very high with a loss of 98% of viable cells after 20 h of incubation. With the addition of $200\;{\mu}g/mL$ of each polysaccharide isolate to the cell line containing $80\;{\mu}g/mL$ of 7-ketocholesterol, polysaccharide isolates from both Flammulina velutipes and Peurotus ostreatus could significantly inhibit the 7-ketochoelsterol-induced cytotoxicity in the cells. But other polysaccharide isolates were not effective in inhibiting cell damage caused by the oxLDL-induced cytotoxicity.

• Journal of Food Hygiene and Safety
• /
• v.23 no.1
• /
• pp.85-89
• /
• 2008

The oxidative stability of refrigerated precooked pork patties containing commercial ${\gamma}$-oryzanoland ${\alpha}$-tocopherol was evaluated. Precooked pork patties containing either ${\gamma}$-oryzanolor ${\alpha}$-tocopherol showed higher oxidative stability (p<0.05) during storage at $4^{\circ}C$ than did the precooked pork patties without the additives (control). The thiobarbituric acid-reactive substances (TBARs) values and warmed-over flavor (WOF) of the precooked pork patties containing ${\gamma}$-oryzanolor ${\alpha}$-tocopherol were lower (p<0.05) than those of the control during refrigerated storage (0, 1, 4, and 8 days). The correlation between TBARs and WOF values was significant (p<0.05). 7-Ketocholesterol content was lower (p<0.05) than those of the control during refrigerated storage (0, 6, 12, 18, and 24 days). The correlation between TBARs values and 7-ketocholesterol content was also significant (p<0.05).

• Journal of Life Science
• /
• v.32 no.2
• /
• pp.142-147
• /
• 2022

Oxysterols are known to be involved in the physiopathology of atherosclerosis. Since 7-ketocholesterol (7-KC) is found in large amounts in oxysterols and in atherosclerotic plaque, the study on how 7-KC may affect monocyte differentiation induced by phorbol myristate acetate (PMA) in the monocytic cell line, THP-1, is essential. 7-KC induced a dose-dependent reduction in cell proliferation without inducing cytotoxicity, and the substantial staining of Nile red demonstrates the increased absorption of intracellular lipids. Although 7-KC itself did not increase cell adhesion, it markedly decreased the adhesion of cells treated with PMA. Furthermore, by observing the effect of 7-KC on phagocytosis using fluorescent-labeled latex beads, 7-KC's ability to abolish phagocytosis in PMA-stimulated macrophages was illustrated. The effect of 7-KC on the expression of selected protein markers on the process of differentiation induced by PMA in THP-1 cells was also examined. 7-KC inhibited expression of ICAM-1, CD11a, SR-A1, and SR-B2 (CD36) in PMA-stimulated THP-1 cells. Conversely, 7-KC drastically increased the expression of SR-D1 (CD68)in PMA-stimulated THP-1 cells. In conclusion, these results suggest that 7-KC modulates monocyte differentiation and activation via the intracellular accumulation of lipid droplets.

• Korean Journal of Food Science and Technology
• /
• v.30 no.6
• /
• pp.1312-1320
• /
• 1998

Some commercial beef loins in raw state were packaged with PVDC as aerobic and vacuum condition. The other beef samples were cooked until core temperature arrived at $70^{\circ}C$ and then packaged immediately in the same way as the raw state. These samples were irradiated by electron beam (0, 1, 2 kGy), and then stored in refrigerator $(2{\sim}4^{\circ}C)$. Identity and quantity of cholesterol oxides were analysed at the 0, 7th, 14th day of storage. In the samples that were raw and packaged aerobically, $7{\alpha}-hydroxycholesterol,\;{\beta}-epoxide,\;7{\beta}-hydroxycholesterol$ and 7-ketocholesterol were detected over $0.5\;{\mu}g/g$. Cholestanetriol and${\alpha}-epoxide$ were detected at levels below $0.5\;{\mu}g/g$ during storage. In the samples that were raw and vacuum-packaged, $7{\alpha}-hydroxycholesterol$, 7-ketocholesterol and cholestanetriol were detected. In the samples that were cooked and packaged aerobically, cholestanetriol and ${\alpha}-epoxide$ were detected below $0.5\;{\mu}g/g$ during storage. $7{\alpha}-hydroxycholesterol,\;{\beta}-epoxide,\;7{\beta}-hydroxycholesterol$and 7-ketocholesterol were detected as $1.53{\sim}26.81,\;1.07{\sim}5.23,\;40.64{\sim}101.30\;and\;7.16{\sim}33.91\;{\mu}g/g$, respectively. In all results, total amounts of cholesterol oxide increased significantly as irradiation dose and storage time increased (P<0.05).

• Journal of Life Science
• /
• v.26 no.2
• /
• pp.226-233
• /
• 2016

Vascular smooth muscle cell (VSMC) apoptosis has been identified in various vascular diseases, including atherosclerosis and restenosis after angioplasty, and has been known to precipitate atherosclerotic plaque instability and rupture. Oxysterols are known as inducers of apoptosis in VSMC, and 7-ketocholesterol (7KC) is the major nonenzymically formed oxysterol in atherosclerotic lesions. The precise mechanism underlying VSMC apoptosis is still poorly understood. In this study, we investigated whether 7KC causes apoptosis, and characterized its apoptotic mechanisms in primary cultured rat aortic VSMC. Cell viability was assessed by MTT assay and trypan blue assay. Apoptosis was assessed by flow cytometry, immunofluorescence, immunoprecipitation, and Western blot analyses. 7KC markedly decreased the VSMC viability in a time- and concentration-dependent manner, and increased the production of 4-hydroxynonenal (HNE), a major end-product of lipid peroxidation, which also decreased the VSMC viability. Pretreatment with 2,4-dinitrophenylhydrazine, a well-known reagent of lipid peroxidation-derived aldehydes, significantly restored the 7KC-decreased viability of VSMC. Furthermore, HNE, as well as 7KC, reduced the level of total Akt, a major mediator of cell survival. The 7KC-decreased level of total Akt was significantly restored by pretreatments with 2,4-dinitrophenylhydrazine and N-acetylcysteine. Lactacystin, a proteasome inhibitor, protected VSMC against apoptosis and Akt degradation, but did not inhibit HNE production. In the immunoprecipitation assay, 7KC increased HNE-modified Akt. From the results, it seems that, in atherosclerotic lesions, 7KC induces HNE production in VSMC, and this HNE binds to Akt, proceeding to proteasomal degradation of Akt, through which mechanism the atherosclerotic plaque instability may be facilitated.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.26 no.2
• /
• pp.261-269
• /
• 1997

The autoxidation of cholesterol and lipid was investigated in mackerel during its salting for 50 days. Furthermore, the effects of antioxidants such as ascorbic acid and BHA on their autoxidation were studied. The cholesterol of mackerel during salting was continuously decreased. Its content was quantified by 23.3mg/100g in salted control sample after 50 days and that is only about 33% of total cholesterol content in fresh mackerel. The addition of BHA in mackerel during salting inhibited cholesterol oxidation more effectively than ascorbic acid.7-Ketocholesterol, unique cholesterol oxidation products was detected in this experiment and malonaldehyde, one of lipid oxidation products, contineuosly increased in control sample all the salting days by the almost same pattern but in the additive samples of ascorbic acid or BHA by different patterns, respectively. BHA was more effective antioxidant against cholesterol and lipid autoxidation than ascorbic acid.

• Journal of Food Hygiene and Safety
• /
• v.23 no.1
• /
• pp.1-5
• /
• 2008

The inhibition of cholesterol autoxidation by commercial ${\gamma}$-Oryzanol (0, 50, 100, and 300 ppm) was studied in an aqueous model system for 20 h at pH 5.5 and $80^{\circ}C$. The inhibition effectiveness of the commercial ${\gamma}$-Oryzanol was followed by the retention of cholesterol and the formation of C-7 oxidized cholesterol derivatives (OCDs). Changes in the amount of ${\gamma}$-Oryzanol in the aqueous system were determined during cholesterol autoxidation. A method to detect the levels of 7-ketocholesterol, $7{\alpha}$-hydroxycholesterol and $7{\beta}$-hydroxycholesterol in an aqueous model system with ${\gamma}$-Oryzanol was developed by using the hexane-ethyl acetate extraction system and high-performance liquid chromatography. Results showed that the levels of C-7 OCDs in an aqueous dispersion containing 300 ppm of ${\gamma}$-Oryzanol were not significantly (p>0.05) increased, when compared to other treatments (0, 50, and 100 ppm), during the accelerated cholesterol oxidation.

• Korean Journal of Food Science and Technology
• /
• v.31 no.1
• /
• pp.74-82
• /
• 1999

Beef, pork and chicken meat that retailed in market were used as experimental samples. Some samples in raw state were packaged with PVDC as aerobic and vacuum condition. The other samples were cooked until internal temperature arrived at $70^{\circ}C$ using electric oven and then packaged immediately in the same way of raw samples. After these samples were irradiated by electron beam (0, 1, 2 kGy), irradiated samples were stored in refrigerator $(2{\sim}4^{\circ}C)$. Identification and quantification of cholesterol oxides were analysed at 0, 7, 14 days. The results were following. The results indicated that raw-vacuum packaged lower detected than that of other treatments. In raw-vacuum packaged, the amounts of $7{\beta}-hydroxycholesterol$ were detected slightly $(below\;0.5\;{\mu}ug/g)$ during storage, and 7-ketocholesterol were detected during every stored time and amounts of this detection were $8.02{\sim}101.30\;{\mu}g/g$. In cooked-aerobic packaged, total amounts of detection were higher than that of other treatments, total amounts of cholesterol oxides were detected about $51.18{\sim}155.90\;{\mu}g/g$ during storage. In all results, pork and chicken samples were similar to the results of beef samples. In all results, total amounts of cholesterol oxides increased significantly as irradiation dose and storage time increased (P<0.05).

• Natural Product Sciences
• /
• v.25 no.1
• /
• pp.49-58
• /
• 2019

Eleven steroid hormones (SHs: androstene-3,17-dione, estrone, ${\beta}$-estradiol, ${\alpha}$-estradiol, testosterone, dehydroepiandrosterone, $17{\acute{a}}$-hydroxyprogesterone, medroxyprogesterone, megestrol acetate, progesterone, and androsterone) were detected from New Zealand deer (Cervus elaphus var. scoticus) velvet antler (NZA, 鹿茸 ). A method for the quantification of eleven SHs was established by using ultraperformance liquid chromatography (UPLC)-MS/MS. The linearities ($R^2$ > 0.991), limits of quantification (LOQ values, 0.3 ng/mL to 23.1 ng/mL), intraday and interday precisions (relative standard deviation: RSD < 2.43%), and recovery rates (97.3% to 104.6%) for all eleven SHs were determined. In addition, a method for the quantification of three 7-oxycholesterols (7-O-CSs: 7-ketocholesterol, $7{\alpha}$-hydroxycholesterol, and $7{\beta}$-hydroxycholesterol) in the NZA was established by using an HPLC-photodiode array (PDA) method. The linearities ($R^2$ > 0.999), LOQ values (30 ng/mL to 350 ng/mL), intraday and interday precisions (RSD < 1.93%), and recovery rates (97.2% to 103.5%) for the three 7-O-CSs were determined. These quantitative methods are accurate, precise, and reproducible. As a result, it is suggested that the five steroid compounds of androstene-3,17-dione, androsterone, 7-ketocholesterol, $7{\alpha}$-hydroxycholesterol, and $7{\beta}$-hydroxycholesterol could be marker steroids of NZA. These methods can be applied to quantify or standardize the marker steroids present in NZA.