• Applied Microscopy
• /
• 제18권2호
• /
• pp.102-118
• /
• 1988

This study was carried out to examine in vitro first whether the storage proteins, which the fat bodies of last larvae from Hyphantria cunea secrete into haemolymph, can be uptaked by the fat body cells of prepupa and then how the uptaked storage proteins can be accumulated in the fat body cells, if uptaken. The fat bodies which had been isolated from last instar larvae were cultured in 1 ml of Grace's insect medium containing $50{\mu}l$ of $^{3}H$-leucine (5.0 mCi/mol, Dupont) at $28{\pm}2^{\circ}C$ for 6 hrs. After the homogenates of the cultured fat bodies were centrifuged at 10,000 rpm for 10 minutes, the proteins included in the supernatant were separated by polyacrylamide gel electrophoreses (non-SDS, 6%). The next treatment of the electrophoresed gel was followed by rinsing. A storage protein band of several bands in the rinsed gel was sliced off. With elution of sliced storage protein bands in Tris-glycine buffer, the purification of radioactive storage proteins from fat bodies was finished. After the purified radioactive storage proteins were added in Grace's insect midis containing fat bodies of the prepupae, they were cultured for the randomly following minutes given as 3, 5, 7, 10, 15, 20 and 30 and for the randomly following hours given as 1, 2, 3 and 4 respectively. The double fixations of the cultured fat bodies in aldehyde and $OsO_4$, were followed by preparation of ultrathin sections from Epon-Araldite blocks through dehydration and embedding. The electron microscope autoradiographic treatment of all prepared sections were performed by the dipping method (Kim et al., 1987). The finally prepared specimens were examined with electron microscope. The fat body cells of the prepupa could be found to uptake the storage preteins of the last instar larvae, which were included in the culture medium, mostly by formation of coated vesicles. The in vitro uptake of the storage proteins actively occurred by 30 minutes after the addition of purified storage proteins in the culture medium. After culture for 7 minutes with the storage proteins, the uptaked radioactive storage proteins labelled a number of lysosomal granules. After culture for 20 minutes with the storage proteins, the radioactive storage proteins were finally incorporated and accumulated in lipid droplets and protein granules. The frequency in the fat body cell of radiolabelled lipid droplets occurs approximately 60%, while the frequency, in which the radiolabelled protein granules occurs in a fat body cell, is approximately 40%.

• Preventive Nutrition and Food Science
• /
• 제8권1호
• /
• pp.40-45
• /
• 2003

Acanthopanax senticosus was grown by a novel, proprietary method, of culturing isolated cells in a bioreactor. An extract from the cells was evaluated for its effect on lipid metabolism in rats fed a high fat diet. Male Sprague-Dawley rats (n＝6) were fed either an AIN-76 diet (control, NDCon), control diet plus Acanthopanax senticosus extract (ND＋Ex), a modified AIN-76 diet supplemented with 20% beef tallow (high fat, HFCon), or a high fat diet plus Acanthopanax senticosus extract (HF＋Ex), for 5weeks. Body weight gain was significantly higher in the HFCon group than the NDCon group. Feed consumption was significantly lower, but energy intake higher, in the groups fed high fat diets compared with the groups fed control diets. Serum HDL-cholesterol concentrations were significantly increased but serum LDL-cholesterol concentrations were decreased in the groups fed the Acanthopanax senticosus extract. Abdominal fat accumulation and serum leptin levels were significantly higher in the HFCon group than the other groups. Carnitine palmitoyltransferase-I (CPT-I) mRNA levels were increased in the groups fed Acanthopanx senticosus extract. These results suggest that supplementation of cell cultured Acanthopanax senticosus extract regulates CPT-I mRNA levels in liver and has an effect on the normalization of lipids in rats fed a high fat diet.

• 한방재활의학과학회지
• /
• 제30권4호
• /
• pp.41-53
• /
• 2020

Objectives The purpose of this study was to investigate the effects of Silbi-san on the antioxidant and fat accumulation inhibition and to analyze the anti-obesity effect by analyzing the changes in serum lipid composition in obese mice. Methods We compared contents of phytochemicals like total polyphenols and total flavonoid and antioxidant activities such as 2,2-dipheny-1-picrylhydrazyl and 2.2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activity. After Silbi-san in 3T3-L1 cells in vitro and mouse adipose tissue ex vivo, we quantified intracellular triglyceride accumulation and lipolysis. Moreover, the anti-obesity activity though inhibiting pancreatic lipase were analyzed. In 3T3-L1 cells, morphological changes showed that control cells had many lipid while cells treated with Silbi-san had less lipid accumulation. 30% EtOH Silbi-san treatment also suppressed the fat absorption by inhibiting the activity of pancreatic lipase and led to high lipolysis through promoting glycerol release. The experimental group was divided into four groups: Normal group fed normal feed, Control group fed 60% high fat diet (HFD) and distilled water, drug group fed 60% high fat diet and 200 mg/kg of Silbi-san water extract, drug group fed 60% HFD and 200 mg/kg of Silbi-san 30% ethanol extract. Results Serum total cholesterol content and serum low density lipoprotein-cholesterol content were significantly decreased in the Silbi-san extract group compared to the control group, serum high density lipoprotein-cholesterol content was significantly increased in Silbi-san extract group. Conclusions In this study, the antioxidant and fat accumulation inhibitory effects of Silbi-san were confirmed.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
• /
• pp.29-34
• /
• 2003

Visceral fat accumulation is known to be an evident clinical index for the insulin resistance related with obesity. Patients with excessive accumulation of visceral fat frequently suffered from metabolic disorder, such as hyperlipidemia, hypertension, and glucose intolerance. However, molecular mechanism for the pathogenesis of obesity-accompanied metabolic disorders has not been fully elucidated. It has been clarified that adipocytes in visceral fat area have different functions from subcutaneous fat area, and these differences might contribute the pathological significance of excessive accumulation of visceral fat for the accompanied insulin resistance and hyperinsulinemia.

• 생명과학회지
• /
• 제10권1호
• /
• pp.22-36
• /
• 2000

The mammary gland contains a subpopulation of epithelial cells with large proliferative potentials which are the likely targets for carcinogens. These clonogenic cells can proliferate and differentiate into functional glandular structures. Multicellular secretory alveolar units (AU) develop from these clonogens in grafts of monodispersed rat mammary epithelial cells (RMEC) in gland-free mammary fat pads in intact recipient F344 rats co-grafted with mammotropic hormone-secreting pituitary tumors (MtT F4). Multicellular nonsecretory ductal units (DU) develop in grafts of monodispersed RMEC in gland-free fat pads in adrenalectomized recipient WF rats co-grafted with MtT W10. However, this effect were reversed by hydrocortisone replacement therapy. RMEC were isolated from appropriate donor rats as monodispersed mixed cells or, alternatively, RNA＋ cells were sorted by flow cytometry of mixed RMEC stained with FITC-RNA and PE-anti-Thy-1.1 monoclonal antibody. We grafted mixed or sorted PNA+ cells in gland-free mammary fat pads in recipient rats that were endocrinologically manipulated to induce AU or DU. Cells were also isolated from these AU or DU as mixed or sorted RNA＋ cells and sub-transplanted in recipient rats treated appropriately to induce AU or DU, respectively. Cells obtained from AU in grafts gave rise to clonal AU and from DU in grafts to DU on sub-transplantation in appropriate recipients. When adrenalectomized recipient WF rats co-grafted with MtT W10 received daily subcutaneous injections of hydrocortisone for periods of 21 days following the PHA＋ cell transplantation, AU, instead of DU, were developed. The histologies of these secondary AU and DU were not different from those of the primary AU and DU. Casein and laminin proteins were demonstrated by immunocytochemical staining of primary and secondary AU. Electron micrographs also demonstrated that AU were composed of secretory cells with milk protein in the cytoplasm. DU were composed of little or non-secretory ductal epithelial cells. These AU and DU also secreted large amounts of lipids. Clonogenic cells were more common in DU than in AU. Thus, AU and DU contain persistent subpopulations of clonogenic stem-like cells.

• Journal of Nutrition and Health
• /
• 제55권1호
• /
• pp.47-58
• /
• 2022

Purpose: Obesity and a high-fat diet (HFD) are risk factors for colorectal cancer. We have previously shown that luteolin (LUT) supplementation in HFD-fed mice markedly inhibits tumor development in chemically induced colon carcinogenesis. In this study, we evaluated the anticancer effect of LUT in the inhibition of cell proliferation in HFD-fed obese mice and HT-29 human colorectal adenocarcinoma cells grown in an adipocyte-derived medium. Methods: C57BL/6 mice were fed a normal diet (ND, 11.69% fat out of total calories consumed, n = 10), HFD (40% fat out of total calories consumed, n = 10), HFD with 0.0025% LUT (n = 10), and HFD with 0.005% LUT (n = 10) and were subjected to azoxymethane-dextran sulfate sodium chemical colon carcinogenesis. All mice were fed the experimental diet for 11 weeks. 3T3-L1 preadipocytes and HT-29 cells were treated with various doses of LUT in an adipocyte-conditioned medium (Ad-CM). Results: The weekly body weight changes in the LUT groups were similar to those in the HFD group; however, the survival rates of the LUT group were higher than those of the HFD group. Impaired crypt integrity of the colonic mucosa in the HFD group was observed to be restored in the LUT group. The colonic expression of proliferating cell nuclear antigen and insulin-like growth factor 1 (IGF-1) receptors were suppressed by the LUT supplementation in the HFD-fed mice. The LUT treatment (10, 20, and 40 µM) inhibited the proliferation and migration of HT-29 cells cultured in Ad-CM in a dose-dependent manner, as well as the differentiation of 3T3-L1 preadipocytes. Conclusion: These results suggest that the anticancer effect of LUT is probably due to the inhibition of IGF-1 signaling and adipogenesis-related cell proliferation in colon cancer cells.

• International Journal of Stem Cells
• /
• 제16권4호
• /
• pp.406-414
• /
• 2023

Dedifferentiated fat cells (DFATs) isolated from mature adipocytes have a multilineage differentiation capacity similar to mesenchymal stem cells and are considered as promising source of cells for tissue engineering. Bone morphogenetic protein 9 (BMP9) and low-intensity pulsed ultrasound (LIPUS) have been reported to stimulate bone formation both in vitro and in vivo. However, the combined effect of BMP9 and LIPUS on osteoblastic differentiation of DFATs has not been studied. After preparing DFATs from mature adipose tissue from rats, DFATs were treated with different doses of BMP9 and/or LIPUS. The effects on osteoblastic differentiation were assessed by changes in alkaline phosphatase (ALP) activity, mineralization/calcium deposition, and expression of bone related genes; Runx2, osterix, osteopontin. No significant differences for ALP activity, mineralization deposition, as well as expression for bone related genes were observed by LIPUS treatment alone while treatment with BMP9 induced osteoblastic differentiation of DFATs in a dose dependent manner. Further, co-treatment with BMP9 and LIPUS significantly increased osteoblastic differentiation of DFATs compared to those treated with BMP9 alone. In addition, upregulation for BMP9-receptor genes was observed by LIPUS treatment. Indomethacin, an inhibitor of prostaglandin synthesis, significantly inhibited the synergistic effect of BMP9 and LIPUS co-stimulation on osteoblastic differentiation of DFATs. LIPUS promotes BMP9 induced osteoblastic differentiation of DFATs in vitro and prostaglandins may be involved in this mechanism.

• 대한한의학회지
• /
• 제28권1호통권69호
• /
• pp.211-223
• /
• 2007

Objectives : This study was performed to investigate the effects of Rhizoma Arisaematis water extract on lipid and glucose metabolism and histochemical change of obese rats. Methods and materials : 10 rats were divided into normal, control and RA (Rhizoma Arisaematis) groups. We fed the control group a high-fat diet and administered normal saline for 8 weeks. We fed the experimental group of rats a high-fat diet and administered an extract of Rhizoma Arisaematis for 8 weeks. At the end of the experiment, the rats were sacrificed to determine their chemical composition. The groups were examined for effects on blood serum lipids, blood sugar, blood insulin concentration and epididymal fat cells. Results : 1. Serum total cholesterol, triglyceride, HDL-cholesterol and glucose of the RA group decreased compared with those of the control group. These decreased rates were significant(P<0.05). 2. Serum LDL-cholesterol, total lipid, free fatty acid and the average size of epididymal fat cells of the RA group decreased compared with those of the control group. These decreased rates were significant(p<0.01). Conclusions : These results suggest that RA may be used to prevent or cure the obesity iniduced by a high-fat diet.

• 한국식품영양과학회지
• /
• 제23권3호
• /
• pp.380-386
• /
• 1994

흰쥐의 간조직에 미치는 gramoxone 독성과 비타민 C의 완화효과에 대해 조사하였다. Gramoxone처리군에서 간세포의 혼탁종창과 지방변화가 사육기간이 증가함에 따라 현저하고 혼탁종창보다는 지방변화가 더 현저하였으며, 특히 간소엽의 문맥야주변대 간세포에서 지방변화가 심하였다. Kupffer세포의 수도 증가하여 4주 사육군에서 제일 많이 증가하였다. Gramoxone-비타민 C 처리군에서 간세포의 혼탁종창과 지방변화가 현저히 감소하였다. 간세포의 glycogen 함량은 정상대조군에 비해 gramoxone처리군에서 다소 증가하는 경향을 나타내었으며 특히 지방변화가 많이 일어난 간세포에서 glycogen 함량이 더 많았다. Gramoxone-비타민 C 처리군에서 gramoxone처리군에 비해 간세포의 glycogen 함량이 감소하는 경향을 나타내었다.

• Toxicological Research
• /
• 제27권2호
• /
• pp.103-110
• /
• 2011

Lipotoxicity involves pathological alterations to cells and tissues in response to elevated fat levels in blood. Furthermore, this process can disturb both cellular homeostasis and viability. In the current study, the authors show that neural progenitor cells (NPCs) are vulnerable to high levels of palmitic acid (PA) a saturated fatty acid. PA was found to cause cell death associated with elevated reactive oxygen species (ROS) levels, and to reduce NPCs proliferation. To evaluate the lipotoxicity of PA in adult NPCs in the hippocampus, male C57BL/6 mice were divided into two groups and maintained on either a normal diet (ND) or PA-rich high fat diet (HFD) for 2 weeks. Interestingly, short-term PA-rich HFD feeding reduced the survival of newly generated cells in the hippocampal dentate gyrus and hippocampal brain-derived neurotrophic factor levels. These findings suggest PA has a potent lipotoxicity in NPCs and that a PA-rich HFD disrupts hippocampal neurogenesis.