Effects of cinnamyl aldehyde on c-Fos and c-Myc expression in NIH3T3 cells

AIM: Cinnamyl aldehyde (CA) is one alcohol ingredient derived from Cinnamomum cassia,which is widely used in treating chronic skin wound in Chinese medicine with the curative effect of ‘rescuing YANG’.The purpose of the present study was to investigate the expression of c-Fos,c-Myc proteins at different time points in NIH3T3 treated with CA and explore the possible mechanism of promoting cell proliferation by CA.METHODS: MTT assay was used for observing cell proliferation.Expression of c-Fos and c-Myc proteins in NIH3T3 cells were assessed by immunocytochemistry assay.RESULTS: The cell proliferation was promoted obviously when CA concentration was between 8.8×10^-2 μg/L and 8.8×10 μg/L.CA at concentration of 5.5 μg/L significantly induced expression of c-Fos,c-Myc proteins at 2-3 h after the stimulation compared with control group (P<0.01).CONCLUSION: CA increases expression of c-Fos and c-Myc proteins,which may be one of mechanisms for CA to promote NIH3T3 cell proliferation.     

Effect of ERK1/2/c-Fos signal pathway on angiotensin-(1-7) inhibiting proliferation of rat glomerular mesangial cell strain induced by angiotensin Ⅱ

AIM: To investigate the effect of ERK1/2/c-Fos signal pathway during angiotensin-(1-7) inhibiting proliferation of rat glomerular mesangial cell strain (GMCS) induced by angiotensin Ⅱ. METHODS: Rat glomerular mesangial cells (GMC) were co-cultured with angiotensin Ⅱ and different doses of angiotensin-(1-7). The numbers of GMC were evaluated by crystal violet staining. The amounts of p-ERK1/2 and c-Fos expressions were detected by Western blotting. RESULTS: Angiotensin- (1-7) showed its inhibitory effects on GMC number increasing induced by angiotensin Ⅱ as well as the amounts of p-ERK1/2 and c-Fos expressions in a concentration dependent manner. CONCLUSION: ERK/c-Fos signal pathway is involved in the inhibitory effects of angiotensin-(1-7) on angiotensin Ⅱ -induced GMC proliferation.     

c-Fos参与羊驼小肠黏膜淋巴细胞生发中心的调控

【目的】研究转录因子c-Fos与凋亡相关蛋白Caspase3在羊驼小肠黏膜淋巴组织的表达,探索c-Fos与羊驼小肠黏膜淋巴细胞生发中心的关系。【方法】用免疫组织化学技术研究c-Fos蛋白与Caspase3蛋白在羊驼十二指肠和空肠淋巴组织的表达。【结果】免疫组织化学结果显示羊驼十二指肠黏膜淋巴细胞较少,c-Fos蛋白主要表达于空肠黏膜下淋巴组织生发中心,在黏膜下淋巴组织向固有层淋巴组织过渡区域有散在表达,在固有层弥散淋巴组织中c-Fos阳性表达细胞较少。Caspase3蛋白主要表达于空肠黏膜固有层淋巴组织,在黏膜下淋巴组织生发中心很少表达,在黏膜下淋巴组织向固有层弥散淋巴组织过渡区域Caspase3有散在表达。【结论】c-Fos参与羊驼空肠黏膜下淋巴组织生发中心增殖与分化的调控。     

Immunolocalization of c-Fos,ELOVL5 and oestradiol in the ewe vulva in relation to oestrus behaviour after treatment with lipopolysaccharide

Sudden activation of the stress axis by a lipopolysaccharide endotoxin (LPS) significantly reduces ewes' sexual attractivity to rams by delaying all signs of oestrous behaviour. To understand mechanisms involved in attracting male interest, we examined c-Fos (nuclear activation), ELOVL5 (enzyme involved in pheromone synthesis) and oestradiol receptors (ER) using immunohistochemistry on ewe vulval tissue at 0, 31 and 40 hr in the ovarian follicular phase with or without exposure to LPS at 28 hr (5 groups of 4 ewes per group). While there was intense staining for immunoreactive (IR)-c-Fos and IR-ELOVL5 in the vulval epithelium and sebaceous glands, there were no differences in intensity between groups of ewes. The absence of IR-ER staining in vulval epithelium and sebaceous/sweat glands was unexpected. Differences in ram behaviour towards ewes in the ovarian follicular phase and after LPS treatment do not appear to involve quantitative changes in vulval c-Fos, ELOVL5 or ER, but subtle qualitative differences in individual-specific compounds (attraction pheromones) remain an option.     

Intragastric gavage with denatonium benzoate acutely induces neuronal activation in the solitary tract nucleus via the vagal afferent pathway

Natural toxic substances have a bitter taste and their ingestion sends signals to the brain leading to aversive oral sensations. In the present study, we investigated chronological changes in c-Fos immunoreactivity in the nucleus tractus solitarius (NTS) to study the bitter taste reaction time of neurons in the NTS. Equal volumes (0.5 mL) of denatonium benzoate (DB), a bitter tastant, or its vehicle (distilled water) were administered to rats intragastrically. The rats were sacrificed at 0, 0.5, 1, 2, 4, 8, or 16 h after treatment. In the vehicle-treated group, the number of c-Fos-positive nuclei started to increase 0.5 h after treatment and peaked 2 h after gavage. In contrast, the number of c-Fos-positive nuclei in the DB-treated group significantly increased 1 h after gavage. Thereafter, the number of c-Fos immunoreactive nuclei decreased over time. The number of c-Fos immunoreactive nuclei in the NTS was also increased in a dose-dependent manner 1 h after gavage. Subdiaphragmatic vagotomy significantly decreased DB-induced neuronal activation in the NTS. These results suggest that intragastric DB increases neuronal c-Fos expression in the NTS 1 h after gavage and this effect is mediated by vagal afferent fibers.     

Local anaesthetics attenuates spinal nociception and HPA-axis activation during experimental laparotomy in pigs

The effect of local anaesthetics on spinal nociception and activation of the hypothalamic-pituitary-adrenal axis (HPA-axis) was examined in a porcine model of abdominal surgery. A standardised laparotomy without visceral involvement was performed on 24 pigs. One group received a unilateral infiltration of mixed lidocaine and bupivacaine in skin, muscle and peritoneum of the surgical area prior to surgery (n=12), while local anaesthetics were replaced by isotonic saline in a second group (n=12). A sham group was subjected to anaesthesia (n=8), but did not undergo surgery. Two hours after surgery, half of the pigs from each group were perfused with formalin and the spinal cord was taken out for stereological quantification of the total number of Fos-like-immunoreactive (Fos-LI) neurones in the dorsal horn. Surgery with saline gave rise to a significant increase in the number of Fos-LI neurones ipsilaterally (107,001+/-16,548; p<0.001) as well as contralaterally (12,766+/-3,842; p<0.01) compared to the sham group. In animals undergoing surgery with LA, the number of Fos-LI neurones ipsilaterally was not significantly different from the sham group (p=0.78), and was reduced significantly both ipsilaterally (6960+/-1662; p<0.001) and contralaterally (3974+/-1131; p<0.05) compared to the saline group. In the other half of each group, blood samples, for determination of ACTH, cortisol, C-reactive protein and interleukin-6 concentrations, were drawn prior to and at predetermined time-points during and after surgery. Surgery with saline gave rise to dramatic increases in plasma ACTH and cortisol (p<0.01 and p<0.001, respectively) within 15 min of incision. In contrast, no changes from the initial concentrations of ACTH and cortisol were observed in pigs receiving local anaesthetics. No changes in plasma concentrations of C-reactive protein or interleukin-6 were observed in either of the groups. These results indicate that spinal nociception and HPA-axis activation caused by laparotomy in pigs can be attenuated by use of infiltration and incisional local anaesthetics prior to surgery. The present model provides a valuable tool in the evaluation of analgesic treatment during surgery, offering objective measures of both nociception and stress.     

Effects of astragalan on neurotransmitters and expression of c-fos mRNA in hippocampus after ischemic brain injury in rats

AIM: To explore the effects of astragalan (AG) on the neurotransmitters,acetylcholine (ACh), norepinephrine (NE) and 5-hydroxytryptamine (5-HT), and the expression of c-fos mRNA in hippocampus after ischemic brain injury in rats. METHODS: Male Wistar rats (180~220 g) were randomly divided into 10 groups (n=10): sham-operated group (SOG), 3 model groups (MG 1 d, 3 d, 7 d) and 3 low- or high-dose AG treatment groups (L/H-AGTG 1 d, 3 d, 7 d), respectively. The middle cerebral artery of the rats in MG group and AGTG group were blocked by operation to induced brain injury. The cerebral blood vessels of the animals were blocked on day 1, day 2 and day 7, respectively, after the L/H-AGTG were treated with AG (5 mg/kg and 15 mg/kg, ip). The content of ACh,5-HT and NE was determined using their respective ELISA kits, and the expression of c-fos mRNA in the hippocampus homogenate was semiquantitative analyzed by RT-PCR after neurologic impairment (NIP) was scored. RESULTS: AG attenuated the injury in hippocampus by cerebral ischemia in a dose-dependent manner. The content of ACh, 5-HT and NE in L-AGTG 7 d,H-AGTG 3 d and 7 d groups was significantly higher than that in MG group, but was lower in SOG group (P<0.05 or P<0.01). The mRNA expression of c-fos in SOG group was lower than that in MG group (P<0.05 or P<0.01), indicating that reinforcement expression of c-fos mRNA by cerebral ischemia and the expression of downstream genes may be beneficial for protecting the neurons. The mRNA expression of c-fos in H-AGTG 3 d/7 d groups was higher than that in MG group (P<0.05). CONCLUSION: AG attenuates the damage of neurons and improves the functions of hippocampus under the condition of cerebral ischemia/reperfusion by increasing the content of ACh, NA and 5-HT, and the mRNA expression of c-fos in hippocampus.