13 research outputs found
Unexpected Role of Ξ±-Fetoprotein in Spermatogenesis
BACKGROUND: Heat shock severely affects sperm production (spermatogenesis) and results in a rapid loss of haploid germ cells, or in other words, sperm formation (spermiogenesis) is inhibited. However, the mechanisms behind the effects of heat shock on spermatogenesis are obscure. METHODOLOGY/PRINCIPAL FINDINGS: To identify the inhibitory factor of spermiogenesis, experimental cryptorchid (EC) mice were used in this study. Here we show that Ξ±-fetoprotein (AFP) is specifically expressed in the testes of EC mice by proteome analysis. AFP was also specifically localized spermatocytes by immunohistochemical analysis and was secreted into the circulation system of EC mice by immunoblot analysis. Since spermatogenesis of an advanced mammal cannot be reproduced with in vitro, we performed the microinjection of AFP into the seminiferous tubules of normal mice to determine whether AFP inhibits spermiogenesis in vivo. AFP was directly responsible for the block in spermiogenesis of normal mice. To investigate whether AFP inhibits cell differentiation in other models, using EC mice we performed a partial hepatectomy (PH) that triggers a rapid regenerative response in the remnant liver tissue. We also found that liver regeneration is inhibited in EC mice with PH. The result suggests that AFP released into the blood of EC mice regulates liver regeneration by inhibiting the cell division of hepatocytes. CONCLUSIONS/SIGNIFICANCE: AFP is a well-known cancer-specific marker, but AFP has no known function in healthy human beings. Our findings indicate that AFP expressed under EC conditions plays a role as a regulatory factor in spermatogenesis and in hepatic generation
Effects of ethyl-esterization, chain-lengths, unsaturation degrees, and hyperthermia on carcinostatic effect of omega-hydroxylated fatty acids
Aim: To evaluate promotive effect of hyperthermia on the carcinostatic activity of synthesized omega-hydroxy fatty acids (wHFAs) and their ethylesters agaist Ehrlich ascites tumor (EAT) cells. Methods: EAT cells were cultured with either wHFAs or their ethylester derivatives in a water bath at either 37 Β°C or 42 Β°C for 30 min, followed by incubation in a CO2 incubator for 20 or 72 h. Mitochondrial dehydrogenase-based WST-1 assay and trypan blue dye exclusion assay were then conducted after incubation. Morphological changes were observed by scanning electron microscopy (SEM). Results: Omega-HFA having a saturated 16-carbon straight-chain (wH16:0) was the most carcinostatic (at 37 Β°C β viability level: 60.0%; at 42 Β°C β 49.6% (WST-1)) among saturated and unsaturated wHFAs with 12, 15 or 16 carbon atoms, when administrated to EAT cells at 100 Β΅M for 20 h. Carcinostatic activity was markedly enhanced by ethyl-esterization of saturated fatty acids, such as wH16:0 (at 37 Β°C β 42.3%; at 42 Β°C β 11.2% , ibid) and wH15:0 (at 37 Β°C β 74.6%; at 42 Β°C β 25.3% , ibid), and their unsaturated counterparts were extremely effective only in combination with hyperthermia. Prolongation of the incubation period to 72 h at the same concentration increased appreciably their carcinostatic effect (wH16:0 ethylesther: 1.3%; wH15:0 ethylesther: 8.0%). These values were also supported by dye exclusion assay. The carcinostatic activity enhanced more markedly by hyperthermia (1.2%; 2.1%, ibid). SEM shows that wH16:0 ethylester-exposed EAT cells underwent extensive injury, such as deformation of cell structure or disappearance of microvilli. Conclusions: wH16:0 ethylester possesses high carcinostatic activity in vitro in combination with hyperthermia and may be utilized as potent anticancer therapeutic agent.Π¦Π΅Π»Ρ: ΠΏΡΠΎΠ°Π½Π°Π»ΠΈΠ·ΠΈΡΠΎΠ²Π°ΡΡ ΡΡΠΈΠ»ΠΈΠ²Π°ΡΡΠΈΠΉ ΡΡΡΠ΅ΠΊΡ Π³ΠΈΠΏΠ΅ΡΡΠ΅ΡΠΌΠΈΠΈ Π½Π° ΠΊΠ°Π½ΡΠ΅ΡΠΎΡΡΠ°ΡΠΈΡΠ΅ΡΠΊΡΡ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΡΠΈΠ½ΡΠ΅Π·ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
ΠΎΠΌΠ΅Π³Π°Π³ΠΈΠ΄ΡΠΎΠΊΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
ΠΆΠΈΡΠ½ΡΡ
ΠΊΠΈΡΠ»ΠΎΡ (HFAs) ΠΈ ΠΈΡ
ΡΡΠΈΠ»ΠΎΠ²ΡΡ
ΡΡΠΈΡΠΎΠ² ΠΏΠΎ ΠΎΡΠ½ΠΎΠ΅Π½ΠΈΡ ΠΊ ΠΊΠ»Π΅ΡΠΊΠ°ΠΌ Π°ΡΡΠΈΡΠ½ΠΎΠΉ ΠΎΠΏΡΡ
ΠΎΠ»ΠΈ ΡΠ»ΠΈΡ
Π°
(EAT). ΠΠ΅ΡΠΎΠ΄Ρ: ΠΊΠ»Π΅ΡΠΊΠΈ EAT ΠΈΠ½ΠΊΡΠ±ΠΈΡΠΎΠ²Π°Π»ΠΈ Ρ HFAs ΠΈΠ»ΠΈ ΠΈΡ
ΡΡΠΈΠ»ΡΡΠΈΡΠ½ΡΠΌΠΈ ΠΏΡΠΎΠΈΠ·Π²ΠΎΠ΄Π½ΡΠΌΠΈ Π½Π° Π²ΠΎΠ΄Π½ΠΎΠΉ Π°Π½Π΅ ΠΏΡΠΈ 37 Β° ΠΈΠ»ΠΈ
42 Β° Π² ΡΠ΅ΡΠ΅Π½ΠΈΠ΅ 30 ΠΌΠΈΠ½ Ρ Π΄Π°Π»ΡΠ½Π΅ΠΉΠΈΠΌ ΠΊΡΠ»ΡΡΠΈΠ²ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ Π² 2
ΠΈΠ½ΠΊΡΠ±Π°ΡΠΎΡΠ΅ Π½Π° ΠΏΡΠΎΡΡΠΆΠ΅Π½ΠΈΠΈ 20 ΠΈΠ»ΠΈ 72 Ρ, ΠΏΠΎΡΠ»Π΅ ΡΠ΅Π³ΠΎ Π°Π½Π°Π»ΠΈΠ·ΠΈΡΠΎΠ²Π°Π»ΠΈ
ΠΆΠΈΠ·Π½Π΅ΡΠΏΠΎΡΠΎΠ±Π½ΠΎΡΡΡ ΠΊΠ»Π΅ΡΠΎΠΊ ΠΌΠ΅ΡΠΎΠ΄Π°ΠΌΠΈ Π°Π½Π°Π»ΠΈΠ·Π° WST-1, ΠΎΡΠ½ΠΎΠ²Π°Π½Π½ΠΎΠ³ΠΎ Π½Π° Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΡΡ
Π΄Π΅Π³ΠΈΠ΄ΡΠΎΠ³Π΅Π½Π°Π·, ΠΈ ΠΏΠΎ
Π²ΠΊΠ»ΡΡΠ΅Π½ΠΈΡ ΡΡΠΈΠΏΠ°Π½ΠΎΠ²ΠΎΠ³ΠΎ ΡΠΈΠ½Π΅Π³ΠΎ. ΠΠΎΡΡΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΈΠ΅ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΡ ΠΊΠ»Π΅ΡΠΎΠΊ ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΠ»ΠΈ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΡΠΊΠ°Π½ΠΈΡΡΡΡΠ΅ΠΉ ΡΠ»Π΅ΠΊΡΡΠΎΠ½Π½ΠΎΠΉ
ΠΌΠΈΠΊΡΠΎΡΠΊΠΎΠΏΠΈΠΈ. Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ: ΠΏΡΠΈ ΠΊΡΠ»ΡΡΠΈΠ²Π°ΡΠΈΠΈ ΠΊΠ»Π΅ΡΠΎΠΊ EAT Π² ΠΏΡΠΈΡΡΡΡΡΠ²ΠΈΠΈ 100 M ΡΠΎΠ΅Π΄ΠΈΠ½Π΅Π½ΠΈΠΉ Π² ΡΠ΅ΡΠ΅Π½ΠΈΠ΅ 20 Ρ ΠΎΠΌΠ΅Π³Π°-HFA
Ρ Π½Π°ΡΡΡΠ΅Π½Π½ΠΎΠΉ 16-ΡΠ³Π»Π΅ΡΠΎΠ΄Π½ΠΎΠΉ ΠΏΡΡΠΌΠΎΠΉ ΡΠ΅ΠΏΡΡ (H16:0) ΠΏΡΠΎΡΠ²Π»ΡΠ»ΠΈ Π½Π°ΠΈΠ±ΠΎΠ»Π΅Π΅ Π²ΡΡΠ°ΠΆΠ΅Π½Π½ΡΠΉ ΠΊΠ°Π½ΡΠ΅ΡΠΎΡΡΠ°ΡΠΈΡΠ΅ΡΠΊΠΈΠΉ ΡΡΡΠ΅ΠΊΡ (ΠΏΡΠΈ
37 Β° ΡΡΠΎΠ²Π΅Π½Ρ ΠΆΠΈΠ·Π½Π΅ΡΠΏΠΎΡΠΎΠ½ΠΎΡΡΠΈ ΡΠΎΡΡΠ°Π²ΠΈΠ» 60,0%; ΠΏΡΠΈ 42 Β° 49,6% (WST-1)) ΠΏΠΎ ΡΡΠ°Π²Π½Π΅Π½ΠΈΡ Ρ ΡΠ°ΠΊΠΎΠ²ΡΠΌ Π½Π°ΡΡΡΠ΅Π½Π½ΡΡ
ΠΈ Π½Π΅Π½Π°ΡΡΡΠ΅Π½Π½ΡΡ
HFAs, ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΡ
12, 15 ΠΈΠ»ΠΈ 16 Π°ΡΠΎΠΌΠΎΠ² ΡΠ³Π»Π΅ΡΠΎΠ΄Π°. Π°Π½ΡΠ΅ΡΠΎΡΡΠ°ΡΠΈΡΠ΅ΡΠΊΠ°Ρ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ Π·Π½Π°ΡΠΈΡΠ΅Π»ΡΠ½ΠΎ Π²ΠΎΠ·ΡΠ°ΡΡΠ°Π»Π° ΠΏΡΠΈ
ΡΡΠΈΠ»ΡΡΠ΅ΡΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ Π½Π°ΡΡΡΠ΅Π½Π½ΡΡ
ΠΆΠΈΡΠ½ΡΡ
ΠΊΠΈΡΠ»ΠΎΡ, ΡΠ°ΠΊΠΈΡ
ΠΊΠ°ΠΊ H16:0 (ΠΏΡΠΈ 37 Β° 42,3%; ΠΏΡΠΈ 42 Β° 11,2%, ibid) ΠΈ H15:0
(ΠΏΡΠΈ 37 Β° 74,6%; ΠΏΡΠΈ 42 Β° 25,3% , ibid), Π² ΡΠΎ Π²ΡΠ΅ΠΌΡ ΠΊΠ°ΠΊ ΠΏΡΠΎΠΈΠ·Π²ΠΎΠ΄Π½ΡΠ΅ Π½Π΅Π½Π°ΡΡΡΠ΅Π½Π½ΡΡ
ΠΊΠΈΡΠ»ΠΎΡ Π±ΡΠ»ΠΈ Π²ΡΡΠΎΠΊΠΎΡΡΡΠ΅ΠΊΡΠΈΠ²Π½Ρ
ΡΠΎΠ»ΡΠΊΠΎ Π² ΠΊΠΎΠΌΠ±ΠΈΠ½Π°ΡΠΈΠΈ Ρ Π³ΠΈΠΏΠ΅ΡΡΠ΅ΡΠΌΠΈΠ΅ΠΉ. Π£Π²Π΅Π»ΠΈΡΠ΅Π½ΠΈΠ΅ ΠΏΠ΅ΡΠΈΠΎΠ΄Π° ΠΈΠ½ΠΊΡΠ±Π°ΡΠΈΠΈ ΠΊΠ»Π΅ΡΠΎΠΊ Π΄ΠΎ 72 Ρ ΠΏΡΠΈ ΡΠΎΠΉ ΠΆΠ΅ ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠΈ Π²Π΅ΡΠ΅ΡΡΠ²
ΠΏΡΠΈΠ²ΠΎΠ΄ΠΈΠ»ΠΎ ΠΊ Π·Π½Π°ΡΠΈΡΠ΅Π»ΡΠ½ΠΎΠΌΡ ΡΠ²Π΅Π»ΠΈΡΠ΅Π½ΠΈΡ ΠΈΡ
ΠΊΠ°Π½ΡΠ΅ΡΠΎΡΡΠ°ΡΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ Π΄Π΅ΠΉΡΡΠ²ΠΈΡ (ΡΡΠΈΠ»ΠΎΠ²ΡΠΉ ΡΡΠΈΡ H16:0 1,3%; ΡΡΠΈΠ»ΠΎΠ²ΡΠΉ ΡΡΠΈΡ
H15:0 ethylesther 8,0%), ΠΏΠΎΠ΄ΡΠ²Π΅ΡΠΆΠ΄Π΅Π½Π½ΠΎΠ³ΠΎ Π΄Π°Π½Π½ΡΠΌΠΈ ΠΎΠΊΡΠ°ΡΠΊΠΈ ΡΡΠΈΠΏΠ°Π½ΠΎΠ²ΡΠΌ ΡΠΈΠ½ΠΈΠΌ. ΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΠ΅ Π³ΠΈΠΏΠ΅ΡΡΠ΅ΡΠΌΠΈΠΈ ΡΠ°ΠΊΠΆΠ΅ ΡΡΠΈΠ»ΠΈΠ²Π°Π»ΠΎ
ΠΊΠ°Π½ΡΠ΅ΡΠΎΡΡΠ°ΡΠΈΡΠ΅ΡΠΊΠΎΠ΅ Π΄Π΅ΠΉΡΡΠ²ΠΈΠ΅ ΡΠΎΠ΅Π΄ΠΈΠ½Π΅Π½ΠΈΠΉ (1,2%; 2,1%, ibid). Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ SEM ΠΏΠΎΠΊΠ°Π·Π°Π»ΠΈ, ΡΡΠΎ
ΠΊΠ»Π΅ΡΠΊΠΈ EAT, ΠΈΠ½ΠΊΡΠ±ΠΈΡΠΎΠ²Π°Π½Π½ΡΠ΅ Ρ ΡΡΠΈΠ»ΠΎΠ²ΡΠΌ ΡΡΠΈΡΠΎΠΌ H16:0, ΡΠ°Π·ΡΡΠ°ΡΡΡΡ Ρ Π½Π°ΡΡΡΠ΅Π½ΠΈΠ΅ΠΌ ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΉ ΡΡΡΡΠΊΡΡΡΡ ΠΈ ΠΈΡΡΠ΅Π·Π½ΠΎΠ²Π΅Π½ΠΈΠ΅ΠΌ
ΠΌΠΈΠΊΡΠΎΠ²ΠΎΠ»ΠΎΠΊΠΎΠ½. ΠΡΠ²ΠΎΠ΄Ρ: Π² ΠΊΠΎΠΌΠ±ΠΈΠ½Π°ΡΠΈΠΈ Ρ Π³ΠΈΠΏΠ΅ΡΡΠ΅ΡΠΌΠΈΠ΅ΠΉ ΡΡΠΈΠ»ΠΎΠ²ΡΠΉ ΡΡΠΈΡ H16:0 ΠΏΡΠΎ Π΅Ρ Π²ΡΡΠΎΠΊΡΡ ΠΊΠ°Π½ΡΠ΅ΡΠΎΡΡΠ°ΡΠΈΡΠ΅ΡΠΊΡΡ
Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ in vitro, ΡΡΠΎ Π³ΠΎΠ²ΠΎΡΠΈΡ ΠΎ Π²ΠΎΠ·ΠΌΠΎΠΆΠ½ΠΎΡΡΠΈ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΡ ΡΠΎΠ΅Π΄ΠΈΠ½Π΅Π½ΠΈΡ Π² ΡΠ΅ΡΠ°ΠΏΠΈΠΈ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
Π·Π°Π±ΠΎΠ»Π΅Π²Π°Π½ΠΈΠΉ
Differential gene expression between wild-type and Gulo-deficient mice supplied with vitamin C
The aim of this study was to test the hypothesis that hepatic vitamin C (VC) levels in VC deficient mice rescued with high doses of VC supplements still do not reach the optimal levels present in wild-type mice. For this, we used a mouse scurvy model (sfx) in which the L-gulonolactone oxidase gene (Gulo) is deleted. Six age- (6 weeks old) and gender- (female) matched wild-type (WT) and sfx mice (rescued by administering 500 mg of VC/L) were used as the control (WT) and treatment (MT) groups (n = 3 for each group), respectively. Total hepatic RNA was used in triplicate microarray assays for each group. EDGE software was used to identify differentially expressed genes and transcriptomic analysis was used to assess the potential genetic regulation of Gulo gene expression. Hepatic VC concentrations in MT mice were significantly lower than in WT mice, even though there were no morphological differences between the two groups. In MT mice, 269 differentially expressed transcripts were detected (β₯ twice the difference between MT and WT mice), including 107 up-regulated and 162 down-regulated genes. These differentially expressed genes included stress-related and exclusively/predominantly hepatocyte genes. Transcriptomic analysis identified a major locus on chromosome 18 that regulates Gulo expression. Since three relevant oxidative genes are located within the critical region of this locus we suspect that they are involved in the down-regulation of oxidative activity in sfx mice