Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer and current therapeutic strategies are limited in their effectiveness. The expressions of Rab5 and the M2 tumor-associated macrophage marker CD163 in tissues were detected by Western blot. The migration and invasion of cells were determined using a Transwell assay. The expressions of the exosome markers were evaluated by Western blot. The polarization of human macrophages (THP-1) was determined by incubation of THP-1 cells with conditioned medium or exosomes collected from MDA-MB-231 cells with indicated transfections or by a coculture system of THP-1 and MDA-MB-231 cells. The M1 and M2 macrophage markers were evaluated by qRT-PCR. The expression of Rab5 in TNBC was significantly higher than that in normal breast tissue. Rab5 expressions in triple-negative and luminal A breast cancer were higher than those in other molecular subtypes. Higher CD163 expression was observed in triple-negative breast cancer and in triple-negative and luminal B subtypes. Rab5 knockdown suppressed but Rab5 overexpression promoted the migration and invasion capacity of MDA-MB-231 cells. The levels of CD63 and CD9 in the medium of Rab5 knockdown cells were lower than those in control cells, whereas higher levels of CD63 and CD9 were observed in Rab5 overexpression cells. Rab5 knockdown decreased the excretion but did not alter the diameter of the exosomes. Knockdown of Rab5 facilitated the anti-tumor polarization of macrophages, which was partially reversed by Rab5 overexpression. Therefore, Rab5 is expected to be a potential therapeutic target for triple-negative breast cancer.
Septin4 belong to a family of polymerizing GTP-binding proteins that are required for many cellular functions, such as membrane compartmentalization, vesicular trafficking, mitosis, and cytoskeletal remodeling. Since, Septin4 is expressed specifically in the testis, we aimed to determine the association between Septin4 gene expression with sperm quality, DNA damage, and stress oxidative level in infertile patients. The present study included 60 semen samples that grouped into three groups: normozoospermia (n=20), asthenozoospermia (n=20), astheno-teratozoospermia (n=20). Initially, semen parameters were analyzed by using the World Health Organization protocol. The mRNA expression of Septin4 in sperm was examined using reverse transcription-polymerase chain reaction. Oxidative stress markers, i.e., total antioxidant capacity, superoxide dismutase, catalase, glutathione peroxidase, and malondialdehyde, were determined by ELISA kit. The current study showed a statistically significant highly positive correlation in Septin4 gene expression with sperm motility, normal morphology, viability, capacity, and sperm mitochondrial membrane potential (MMP). However, it showed significant negative correlation with sperm DNA fragmentation. Septin4 had a significant correlation with stress oxidative factor and antioxidant enzyme levels. In conclusion, Septin4 gene expression provides clinical useful information for the diagnosis of male infertility. It might be a marker for discrimination between fertile and infertile patients. The current study showed a statistically significant highly positive correlation in Septin4 gene expression with sperm motility, normal morphology, viability, capacity, and sperm MMP. However, it shows significant negative correlation with sperm DNA fragmentation. Septin4 had a significant correlation with stress oxidative factor and antioxidant enzyme levels.
In rat kidney, the changes in concentrations of nucleotides and their derivatives during ischemia induced by renal artery ligation was measured quantitatively with high performance liquid chromatography(HPLC). After the ligation of renal artery for 60minutes, the concentrations of the nucleotides and derivatives were measured. In ischemic tissue, IDP was significantly decreased from $217.4{\pm}12.68{\mu}g$ in control to $80.7{\pm}18.39{\mu}g$ (p<0.01) ; ATP, $307.2{\pm}56.63{\mu}g$ to $47.6{\pm}5.95{\mu}g$ (p<0.01) ; ADP+AMP, $227.1{\pm}7.98{\mu}g$ to $61.4{\pm}3.92{\mu}g$(P<0.01); $NAD^+$, $217.9{\pm}4.49{\mu}g$ to $126.6{\pm}10.44{\mu}g$(P<0.01) ; GTP, $202.5{\pm}23.76{\mu}g$ to $117.7{\pm}14.24{\mu}g$ (P<0.05) ; GMP, $54.5{\pm}9.03{\mu}g$ to $23.7{\pm}0.46{\mu}g$(p<0.05), and inosine, $16.6{\pm}3.45{\mu}g$ to $7.8{\pm}0.87{\mu}g$ (P<0.05). But hypoxanthine and xanthine were significantly increased from $113.0{\pm}15.58{\mu}g$ to $159.7{\pm}12.07{\mu}g$ (P<0.05) and from $87.7{\pm}6.77{\mu}g$ to $173.1{\pm}12.52{\mu}g$ (P<0.01). In ischemic kidney, concentration of ATP was decreased to 39.9% of control at 10 minutes, 19.8% at 30 minutes, and 15.5% at 60 minutes, and ADP+AMP were decreased to 70.3% of control at 10 minutes, 67.3% at 30 minutes, and to 27.0% at 60 minutes, but hypoxanthine and xanthine were increased to 121.5% and 127.1% at 10 minutes, 126.0% and 174.4% at 30 minutes, and 141.4% and 197.3% at 60 minutes Total adenosine nucleotides were decreased to 20.3% of control during 60 minutes of ischemia, but hypoxanthine and xathine were increased to 157.5 % of control. These results suggest that the changes in the concentration of nucleotides and their metabolic derivatives are useful indices of the extents of tissue ischemia in rat kidney.
Journal of the Korean Society of Food Science and Nutrition
/
v.38
no.9
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pp.1153-1160
/
2009
Monascus-fermented Korean red ginseng (MFRG), fermented by Monascus purpureus KCCM 12002, may be an ideal candidate for the hepatoprotectic, hypolipidemic and antioxidative activities. Effect of MFRG powder on these parameters in rats was investigated. Body weight gain, food intake, and water consumption were not significantly different among the groups. Total and relative weights of liver were significantly higher in MFRG group than that in other groups. The activities of AST and $\gamma$-GTP were highly lowered in MFRG group compared to control group. Contents of serum total lipid and triglyceride were significantly lowered in silymarin group and were significantly increased in MFRG group compared to control group, but tended to be lowered in RG group. Serum content of total cholesterol tended to be lowered in silymarin, RG, and MFRG groups compared to control group. HDL-cholesterol contents was only significantly increased in MFRG group compared to control group. At the same time, atherogenic index (AI) was also significantly lowered in silymarin, RG and MFRG groups compared to control group, and this effect was more pronounced in MFRG group. Content of thiobarbituric acid reactive substances (TBARS) in the liver was significantly lowered in MFRG group and tended to lowered in silymarin and RG groups compared to control group. The hepatic glutathione concentration was significantly higher in silymarin and MFRG groups. Hepatic morphology in all experimental groups revealed clear-cut hepatic lobules with the uniform pattern of the polyhedral hepatocytes radiating towards the periphery from the central vein. These results suggested that MFRG may have anti-atherogenic index (AI) and antioxidative activity in normal dietary feeding rats.
The current study was performed to investigate the effects of dietary supplementation of dried coffee meal (CM) on growth performance, intestinal and blood biochemical index, intestinal enzymes, and cecal microbial populations. A total of 162, 3-day-old male broiler chicks were randomly allocated into three dietary groups: control group (CON), basal diet added with 0.5% CM (CM I), and basal diet added with 1.0% CM (CM II). Dietary supplementation of CM did not change bird performance and the relative weight of intestinal mucosal tissues. The birds fed the diet supplemented with CM (0.5 and 1.0%) significantly decreased mucosal glucose concentration (P<0.05) without affecting blood glucose level compared with those fed control diet. The level of blood aspartate aminotransferase (AST) significantly increased in CM II group (P<0.05) without affecting ${\gamma}$-glutamyl transpeptidase (${\gamma}$-GTP) compared with that in the CON group. The specific activity of intestinal maltase, leucine aminopeptidase (LAP) and alkaline phosphatase (ALP) were not affected by dietary supplementation of CM, whereas sucrase activity in birds fed the diet supplemented with CM was decreased (P<0.05) compared to that in the control birds. The colony forming units (CFU) of E. coli in the cecum of CM-fed birds was significantly decreased (P<0.05) compared with that of control birds without changing the CFU of Lactobacillus. In conclusion, dietary supplementation of lower level of CM (0.5%) can be used as a beneficial feed resource without liver toxicity in broiler chicks.
Intracellular calcium concentration ($[Ca^{2+}]_i$) may play a crucial role in a variety of neuronal functions. Here we report that in primary culture of mouse cerebellar granule cells nicotinic acetylcholine receptors (nAChRs) are expressed in a specific developmental stage and involved in the regulation of intracellular calcium homeostasis. Nicotine-mediated calcium responses were measured using $^{45}Ca^{2+}$ or fluorometrically using the calcium-sensitive fluorescent dye fura-2. Maximal uptake of $^{45}Ca^{2+}$ evoked by nicotine in mouse cerebellar granule cells were revealed $8{\sim}12$ days in culture. In contrast, nicotine did not alter the basal $^{45}Ca^{2+}$ uptake in cultured glial cells. In cerebellar granule cells nicotine-evoked $^{45}Ca^{2+}$ uptake was largely blocked by the NMDA receptor antagonists. Glutamate pyruvate transaminase (GPT). which removes endogenous glutamate, also prevented nicotine effects, implying the indirect involvement of glutamate in nicotine-mediated calcium responses. Fluorometric studies using fura-2 showed two phases of nicotine-evoked $[Ca^{2+}]_i$ rises: the initial rising phase and the later plateau phase. Interestingly, the NMDA receptor antagonists and GPT appeared to inhibit only the later plateau phase of nicotine-evoked $[Ca^{2+}]_i$ rises. The present results imply that nicotine mediated $^{45}Ca^{2+}$ uptake and $[Ca^{2+}]_i$ rises are attributed to the calcium fluxes through both nAchRs and NMDA receptors in a time-dependent manner. Consequently, nAChRs may play an important role in neuronal development by being expressed in a specific developmental stage and regulating the intracellular calcium homeostasis.
In order to determine the effective concentration, 2, 7, and 14 brix black garlic extracts (BGE) were fed to restraint stressed rats and the lipid metabolism was analyzed. The dietary intake of the 14 brix BGE-fed group was $547.64{\pm}13.99$ g/4 weeks and it was significantly higher than that of the control group. The weight of all organs from the all BGE-fed groups showed no significant differences. While BGE concentration increased, BGE-fed groups tend to have low content of blood glucose. It was the lowest in the 14 brix BGE-fed group ($121.32{\pm}3.50$ mg/dl). Total cholesterol content was $54.54{\pm}1.63{\sim}65.84{\pm}2.61$ mg/dl. Total cholesterol content of the 14 brix BGE-fed group was the lowest and, in addition, lower than the normal group. Triglyceride content of serum showed the same tendency. HDL-cholesterol concentrations of the restraint stressed control (RSC) group were significantly higher than those of 7 and 14 brix BGE-fed groups. LDL and VLDL-cholesterol contents showed contrary results. GOT, GTP, and ALP activity of serum were lower when BGE concentration was higher. It is expected that BGE has a liver protective activity. In the liver, total lipids and triglyceride contents were significantly lower in BGE-fed groups than in the ISC group. However, total cholesterol content showed no significant difference between groups. TBARS content in liver of the 2 brix BGE-fed group was not significantly different than control, while 7 and 14 brix BGE-fed groups were lower than control. Glycogen content in the liver was only significantly lower in the 14 brix BGE-fed group than in the RSC group. Findings indicate that feed of BGE over 7 brix contributes to resolving the restraint-stressed rats by improving the lipid metabolism and liver protective activity.
Kang, Min-Jung;Lee, Soo Jung;Sung, Nak Ju;Shin, Jung-Hye
Journal of Life Science
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v.23
no.3
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pp.432-442
/
2013
In this study, we examined the biological activity and synergistic effects of an extract powder of 1% and 3%, each fresh (FGP) and black garlic (BGP) in streptozotocin-induced diabetic rats. Blood glucose content was significantly lower in FGP and BGP groups than control group. Glycosylated Hb was significantly higher in streptozotocin induced diabetic control group than normal group, but significantly lower in FGP and BGP groups. Total cholesterol content of the FGP and BGP groups were lower than control group, but not shown the significant difference between garlic fed groups. HDL-cholesterol concentrations of the FGP and BGP fed groups were significantly higher than control group, except of 1% BGP group. LDL and VLDL-cholesterol contents were significantly lower in 3% FGP group, and the same tendency atherogenic index and cardiac risk factor. GOT, GPT, and ${\gamma}$-GTP activity of serum were lower in FGP and BGP fed groups than control group. Glycogen contents in liver significantly higher than control group, and has not significantly difference between normal group. TBARS content was no significantly difference in the liver and serum, but in the kidney, 3% FPG and BGP fed groups were significantly lower than other experimental group. DPPH radical scavenging activity of liver has not significant difference among experimental groups, but activity was higher garlic extract powder fed groups in serum and kidney. These results indicate that dietary supplements of fresh and black garlic extract powder was contributed to lower of blood glucose, loss prevention of glycogen in liver and improve of lipid metabolism.
In this experiment, three different diets were produced to investigate the effects of Jeju native chicken eggs fed with citrus peel on the profiles of rats. The first diet did not contain any eggs (TS), the second diet contained 10% eggs and no citrus peels (T0), and the third diet contained 10% chicken egg with citrus peels (T1). These diets were provided to 11-week-old male rats for four weeks. The weight gain, feed intake and feed efficiency were not significantly different between the three treatment groups, TS, T0, and T1. The weight of the liver was significantly higher in T0 and T1 than TS (p<0.05), but the weights of the kidney and epididymal fat pad were not significantly different between the TS, T0, and T1 groups. The total lipid, phospholipid, triglycerides, total cholesterol and LDL cholesterol did not significantly difference among the TS, T0, and T1 groups. The HDL cholesterol and HDL cholesterol/ total cholesterol of T1 were the highest among the groups, and the atherogenic index of T1 was the lowest among the groups (p<0.05). The total protein of TS was significantly lower in T0 and T1, and the albumin of T1 was the highest among the groups (p<0.05). The albumin/globulin ratio of TS was significantly higher in T0 and T1, but the creatinine of T0 was significantly higher in T0 and T1 (p<0.05). The blood sugar and blood pigment were not significantly different between the TS, T0, and T1 groups. The $\gamma$-GTP of T1 was the lowest among the groups (p<0.05), but the ALT, AST and ALP did not show significant difference among the TS, T0, and T1 groups.
Studies have shown that onions exhibit a wide variety of health-promoting properties. The health benefits by the onion have been attributed to its ability to scavenge free radicals, to reduce blood lipids, to lower blood pressure, and to inhibit platelet aggregation. This study was performed to investigate whether onion extract supplementation would affect the blood markers of ethanol-induced fatty liver in rats. Initially, male Sprague-Dawley rats were housed singly in a room of controlled temperature and lighting and had free access to a nutritionally adequate AIN-93G and deionized water. The rats were trained for meal feeding to prevent a decline in food intake, as inevitably observed following an ethanol feeding. After the training period, rats were weight-matched and assigned to the following three groups: 1) a control group, fed the AIN-93G diet alone (control); 2) an ethanol group, fed the AIN-93G diet with ethanol at 4 g/day/kg body weight (ethanol); and 3) an onion group, fed the AIN-93G diet with ethanol plus supplemental freeze-dried onion powder at 500 mg/day/rat (ethanol + onion). All three group were meal-fed 7.0 g of their respective diets at 0900 h and 7.5 g at 1600 h for 28 days. At 0, 2, and 4 wk, blood was collected via the orbital sinus and organs were collected following overnight food deprivation. Both control and experimental groups continually gained weight throughout the study. No significant differences in the weights of the liver, kidneys, heart, spleen, and testis were observed. However, the serum level of triglycerides was significantly increased by ethanol but significantly decreased by onion extract. The activities of serum glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) at 4 wk were significantly increased by ethanol feeding but were significantly decreased by onion supplementation. However, no differences among groups were observed in the serum levels of alkaline phosphatase, gamma-glutamyl transpeptidase, bilirubin, and protein. These results provide that onion extract favorably affect alcoholic fatty liver by decreasing the serum concentration of triglyceride and the activities of GOT and GPT.
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