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Taurine exerts neuroprotective effects via anti-apoptosis in hypoxic-ischemic brain injury in neonatal rats (신생 흰쥐의 저산소성 허혈성 뇌손상에서 항세포사멸사를 통한 taurine의 신경보호 효과)

  • Jeong, Ji Eun;Kim, Tae Yeol;Park, Hye Jin;Lee, Kye Hyang;Lee, Kyung Hoon;Choi, Eun Jin;Kim, Jin Kyung;Chung, Hai Lee;Seo, Eok Su;Kim, Woo Taek
    • Clinical and Experimental Pediatrics
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    • v.52 no.12
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    • pp.1337-1347
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    • 2009
  • Purpose:Taurine (2-aminoethanesulfonic acid) is a simple sulfur-containing amino acid. It is abundantly present in tissues such as brain, retina, heart, and skeletal muscles. Current studies have demonstrated the neuroprotective effects of taurine, but limited data are available for such effects during neonatal period. The aim of this study was to determine whether taurine could reduce hypoxic-ischemic (HI) cerebral injury via anti-apoptosis mechanism. Methods:Embryonic cortical neurons isolated from Sprague-Dawley (SD) rats at 18 days gestation were cultured in vitro. The cells were divided into hypoxia group, taurine-treated group before hypoxic insult, and taurine-treated group after HI insult. In the in vivo model, left carotid artery ligation was performed in 7-day-old SD rat pups. The pups were exposed to hypoxia, administered an injection of 30 mg/kg of taurine, and killed at 1 day, 3 days, 1 week, 2 weeks, and 4 weeks after the hypoxic insult. We compared the expressions of Bcl-2, Bax, and caspase-3 among the 3 groups by using real- time polymerase chain reaction (PCR) and western blotting. Results:The cells in the taurine-treated group before hypoxic insult, although similar in appearance to those in the normoxia group, were lesser in number. In the taurine-treated group, Bcl-2 expression increased, whereas Bax and caspase-3 expressions reduced. Conclusion:Taurine exerts neuroprotective effects onperinatal HI brain injury due to its anti-apoptotic effect. The neuroprotective effect was maximal at 1-2 weeks after the hypoxic injury.

Functional recovery after transplantation of mouse bone marrow-derived mesenchymal stem cells for hypoxic-ischemic brain injury in immature rats (저산소 허혈 뇌 손상을 유발시킨 미성숙 흰쥐에서 마우스 골수 기원 중간엽 줄기 세포 이식 후 기능 회복)

  • Choi, Wooksun;Shin, Hye Kyung;Eun, So-Hee;Kang, Hoon Chul;Park, Sung Won;Yoo, Kee Hwan;Hong, Young Sook;Lee, Joo Won;Eun, Baik-Lin
    • Clinical and Experimental Pediatrics
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    • v.52 no.7
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    • pp.824-831
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    • 2009
  • Purpose : We aimed to investigate the efficacy of and functional recovery after intracerebral transplantation of different doses of mouse mesenchymal stem cells (mMSCs) in immature rat brain with hypoxic-ischemic encephalopathy (HIE). Methods : Postnatal 7-days-old Sprague-Dawley rats, which had undergone unilateral HI operation, were given stereotaxic intracerebral injections of either vehicle or mMSCs and then tested for locomotory activity in the 2nd, 4th, 6th, and 8th week of the stem cell injection. In the 8th week, Morris water maze test was performed to evaluate the learning and memory dysfunction for a week. Results : In the open field test, no differences were observed in the total distance/the total duration (F=0.412, P=0.745) among the 4 study groups. In the invisible-platform Morris water maze test, significant differences were observed in escape latency (F=380.319, P<0.01) among the 4 groups. The escape latency in the control group significantly differed from that in the high-dose mMSC and/or sham group on training days 2-5 (Scheffe's test, P<0.05) and became prominent with time progression (F=6.034, P<0.01). In spatial probe trial and visible-platform Morris water maze test, no significant improvement was observed in the rats that had undergone transplantation. Conclusion : Although the rats that received a high dose of mMSCs showed significant recovery in the learning-related behavioral test only, our data support that mMSCs may be used as a valuable source to improve outcome in HIE. Further study is necessary to identify the optimal dose that shows maximal efficacy for HIE treatment.

Neuroprotective effects of resveratrol via anti-apoptosis on hypoxic-ischemic brain injury in neonatal rats (신생 백서의 저 산소 허혈 뇌손상에서 항세포사멸사를 통한 resveratrol의 신경보호 효과)

  • Shin, Jin Young;Seo, Min Ae;Choi, Eun Jin;Kim, Jin Kyung;Seo, Eok Su;Lee, Jun Hwa;Chung, Hai Lee;Kim, Woo Taek
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1102-1111
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    • 2008
  • Purpose : Resveratrol, extracted from red wine and grapes, has an anti-cancer effect, an antiinflammatory effect, and an antioxidative effect mainly in heart disease and also has neuroprotective effects in the adult animal model. No studies for neuroprotective effects during the neonatal periods have been reported. Therefore, we studied the neuroprotective effect of resveratrol on hypoxic-ischemic brain damage in neonatal rats via anti-apoptosis. Methods : Embryonic cortical neuronal cell culture of rat brain was performed using pregnant Sprague-Dawley (SD) rats at 18 days of gestation (E18) for the in vitro approach. We injured the cells with hypoxia and administered resveratrol (1, 10, and $30{\mu}g/mL$) to the cells at 30 minutes before hypoxic insults. In addition, unilateral carotid artery ligation with hypoxia was induced in 7-day-old neonatal rats for the in vivo approach. We injected resveratrol (30 mg/kg) intraperitoneally into animal models. Real-time PCR and Western blotting were performed to identify the neuroprotective effects of resveratrol through anti-apoptosis. Results : In the in vitro approach of hypoxia, the expression of Bax, caspase-3, and the ratio of Bax/Bcl-2, indicators of the level of apoptosis, were significantly increased in the hypoxia group compared to the normoxia group. In the case of the resveratrol-treated group, expression was significantly decreased compared to the hypoxia group. And the results in the in vivo approach were the same as in the in vitro approach. Conclusion : The present study demonstrates that resveratrol plays neuroprotective role in hypoxic-ischemic brain damage during neonatal periods through the mechanism of anti-apoptosis.

Injury of Neurons by Oxygen-Glucose Deprivation in Organotypic Hippocampal Slice Culture (뇌 해마조직 절편 배양에서 산소와 당 박탈에 의한 뇌신경세포 손상)

  • Chung, David Chanwook;Hong, Kyung Sik;Kang, Jihui;Chang, Young Pyo
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1112-1117
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    • 2008
  • Purpose : We intended to observe cell death and apoptotic changes in neurons in organotypic hippocampal slice cultures following oxygen-glucose deprivation (OGD), using propidium iodide (PI) uptake, Fluoro-Jade (FJ) staining, TUNEL staining and immunofluorescent staining for caspase-3. Methods : The hippocampus of 7-day-old rats was cut into $350{\mu}m$ slices. The slices were cultured for 10 d (date in vitro, DIV 10) and and exposed to OGD for 60 min at DIV 10. They were then incubated for reperfusion under normoxic conditions for an additional 48 h. Fluorescence of PI uptake was observed at predetermined intervals, and the cell death percentage was recorded. At 24 h following OGD, the slices were Cryo-cut into $15{\mu}m$ thicknesses, and Fluoro-Jade staining, TUNEL staining, and immunofluorescence staining for caspase-3 were performed. Results : 1) PI uptake was restricted to the pyramidal cell layer and DG in the slices after OGD. The fluorescent intensities of PI increased from 6 to 48 h during the reperfusion stage. The cell death percentage significantly increased time-dependently in CA1 and DG following OGD (P<0.05). 2) At 24 h after OGD, many FJ positive cells were detected in CA1 and DG. Some neurons had distinct nuclei and processes while others had fragmented nuclei and disrupted processes in CA1. TUNEL and immunofluorescent staining for caspase-3 showed increased expression of TUNEL labeling and caspase-3 in CA1 and DG at 24 h after OGD. Conclusion : The numerous dead cells in the slice cultures after OGD tended to display apoptotic changes mediated by the activation of caspase-3.

Effects of Methylprednisolone on TGF-${\beta}1$, Apoptosis and Renal Scarring in Experimental Acute Pyelonephritic Weaning Rats (실험적 급성 신우신염이 유발된 이유기 백서에서 methylprednisolone이 TGF-${\beta}1$, 세포고사 및 신반흔에 미치는 영향)

  • Whang, Soo-Ja;Sung, Soon-Hee;Lee, Seung-Joo
    • Childhood Kidney Diseases
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    • v.6 no.1
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    • pp.75-84
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    • 2002
  • Purpose Acute pyelonephritis of growing kidneys may result in renal scarring. TGF-${\beta}1$, inflammatory cytokine, has been suggested to play an important role in promoting renal scarring through apoptosis, suppression of cellular proliferation and fibrosis. We observed the effects of a potent anti-inflammatory agent, methylprednisolone on apoptosis and renal scarring in experimentally induced acute pyelonephritic weaning rats. Materials and Methods: To induce ascending pyelonephritis a saline solution containing Escherichia coli type ATCC No. 25922, pili- form (107 bacteria/mL) was infused into the bladder through the 16-guage silicone cannula for 48 hours to 102 three-week-old Sprague-Dawley rats (50-60g). Experimental groups were divided into three groups according to the treatment protocols, group I (ceftriaxone only, n=3l), group II (methylprednisolone+ceftriaxone n=28), control group (n=43) was not treated. Histopathologic scores of inflammatory changes, fibrosis and tubular atrophy, the apoptosis index and TGF-${\beta}1$ expression score were observed at post-infection 1 and 3 week. Datas were analysed using ANOVA test and P value below 0.05 was interpreted as significant. Results: The mortality rate ($21.4\%$) of group II was not different to the control group ($41.9\%$) and group I ($32.3\%$). The inflammatory score of group II ($0.8{\pm}0.87$) at week 1 was significantly lower than those of the control group ($2.3{\pm}0.87$) and Group I ($1.7{\pm}0.79$) (P<0.05). Apoptosis index of group II ($2.9{\pm}2.15$) at week 1 was significantly lower than those of the control group ($10.0{\pm}1.95$) and group 1 ($8.3{\pm}2.53$) (P<0.05). TCF-${\beta}1$ expression score of group II ($0.8{\pm}0.72$) at week 1 was significantly lower than those of the control group ($1.9{\pm}0.68$) and group I ($1.8{\pm}0.60$) (P<0.05). The fibrosis score of group II ($1.1{\pm}1.10$) at week 3 was significantly lower than that of the group I ($1.8{\pm}0.83$) (P<0.05) Conclusion: Conclusion Combined treatment with methylprednisolone and ceftriaxone reduced inflammation, fibrosis, apoptosis and TGF-${\beta}1$ expression in acute pyelonephritic weaning rats, compared to ceftriaxone alone. Anti-inflammatory agent supplemented to antibiotics could prevent renal scarring more effectively. (J Korean Soc Pediatr Nephrol 2002 ; 6 : 75-84)

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A Study on the Actual Conditions of Smoking in Middle and High School Students in One Region (일 지역 중·고등학생의 흡연실태)

  • Kim, Hyeon-Ok
    • Journal of the Korean Society of School Health
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    • v.12 no.1
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    • pp.149-167
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    • 1999
  • To investigate the actual conditions of smoking in middle and high school students in Chinan County, I used a sturctured questionnaire for 1,579 students attending twelve middle-high schools from December 1, 1998 to December 20, 1998. I collected and data correlated the using an $SPSS-PC^+$ 1. The smoking rate of middle-high schoo1 students in Chinan County was 17.9%, relatively high. This smoking rate was different according to the gender, grade, religion, and economic situation. In mals, high school students, non-religious, students low income family students, the smoking rate was higher. The smoking rate of high school students was almost the same as the smoking rate of adults, generally higher than that of foreign teenagers. Because the smoking rat of studinets in the third grade of middle school and in the first grade of high School was six times higher, increased education should be conducted during this time in an attempt to curb the sudden increase of the smoking rate. The smoking rate of girl students was 5.0%; this has increased mor than three times from ten years ago. Consequently, counter measures should be taken against the smoking of female students as well as juvenile smoking in general. In addition, the smoking rate of middle-high school students showed interesting differences when correnated with enviornmental factors. Students with low grades, who are not satisfied with school life, who don't have both parents, who have uncaring parents who nare too strict or too arbitary, who have smoking parents, or who have experienced smoking commonly smoked. Therefore, to lower the smoking rate we should improve the school environment, improve a student's interest in school life. And parents or siblings should lead by example and quit smoking at home. Schools should educate students more effectively concerning the harmful effects of smoking and create an accurate understanding of its dancers. From the beginning, we should teach students never ever to touch cigaretts. 2. The surve discovered that most students started out of curiosity, or solicitantion from friends or elders at middle school, and had been smoking one to five cigarettes for more than a year. They obtained cigarettes at stores and most of them have friends who smoke. As a result anti-smoking education should be conducted at elementary schools prior to middle school. More than 95% of the teenagers who smoke had friends who smote and smoked out of curiosity or the recommendation of elders. Thus, we must focus on teenagers who smoke in group, rather than individually. Fuyrthermore, the strict application of the regulation of tobacco sales as well as tobacco cooperation from retailers are needed. While students did not show any mood or academic achievement difference after beginning smoking, 58.1% of the students a health situdation that was worse. Juvenile smoking is more harmful to the juvenile than adult smoking is to the adult. This should be focused on in an anti-smoking campaign. 3. Students who smoke hada more positive attitude toward smoking than students who don't smoke. Students who smoke had a tendency to have a nuetral position and are not concerned about smoking compared to non-smoking students. The survey showed that the great number of students had a nuetral position. Because this nuetrality may increase Juvenile smoking, education that provides an exact understanding of smoking should be performed to build the correct attidude toward smoking. 4. Middle school students smoke when angry, gloomy, anxious, a lone and when they have some problems to solve, on when they feel inconveniened in other wores, they smoke to reliver stress. They also smok due to addiction. Because smoking is not a praetical method to relieve stress, a program which helps to acquire positive relief stress should be provided to help reduce smoking. 5. About 65% of students who smoke want to quit smoking because of health problem, 78% of them have tried mor than once to quit but failed due to weak will power and peer pressure from friends who smoke. Juvenile smoking is group, oriented. Thus, the program that advances less smoking will be the one that focuseds on groups. 6. As for advice to students who want to quit smoking, "persuasion" was used most commonly, followed by a "presentation on how to quit smoking". Another method were severe punishment. About 70% of the students wanted the anti-smoking guide at school. 7. Most students (73.5%) had a position that more anti-smoking education at school is needed. Obriously, then, anti-smoking education at middle-high schools should be reinfoced. Although the education which explains the harmful influence of tobacco is known as an efficient way prevent smoking; it does not influence students who already smoke. Therefore, for students who smoke, multi-dimensional approaches must be attempted that include physical training, phychokogical approache, consultation and discussion, medical chek-ups, audio-visual education technigues, and professonal instructors, in addition, because smoking students have more negative on lukewarm attitude to anti-smoking education anti-smoking education should be conducted through a communicative style by dedicated teachers who care about students. In order to increase the effectiveness of this program.

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The Effects of Moxifloxacin in Endotoxin-induced Acute Lung Injury (내독소로 유도된 급성폐손상에서 Moxifloxacin의 효과)

  • Baek, Jong Hyun;Lee, Jang Hoon;Lee, Jung Cheul;Lee, Dong Hyup;Moon, Yong Suk;Lee, Young Man
    • Journal of Chest Surgery
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    • v.42 no.1
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    • pp.1-8
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    • 2009
  • Background: The pathophysiology of acute respiratory distress syndrome with sepsis is acute lung injury (ALI) that's' caused by endotoxin (LPS). We evaluate effects of moxifloxacin on LPS-induced ALI in a rat model. Material and Method: The rats were divided into 3 groups as the control group (C), the LPS insult group (L), and the LPS+moxifloxacin treated group (L-M). ALI was induced by endotracheal instillation of E.coli LPS, then moxifloxacin was given in 30 minutes. Five hours later, we checked the lung weight/body weight ratio(the L/BW ratio), the protein & neutrophils in the bronchoalveolar lavage fluid (BALF), the myeloperoxidase (MPO) activity & the malondialdehyde (MDA) content, the expressions of cytosolic and secretory phospholipase $A_2$ (c, $sPLA_2$), and the morphology of the lung with using a light microscope. Result: The L/BW ratio, the protein content and the neutrophil count in the BALF, and the MPO activity and the MDA content in lung were significantly increased in group L compared to group C, and these factors were markedly decreased in group L-M compare to group L. The $cPLA_2$ expression and the $sPLA_2$ expression were increased in group L and the $cPLA_2$ expression was decreased in group L-M. Yet the $sPLA_2$ expression was not changed in group L-M. Morphologically, many inflammatory findings were observed in group L, but not in group L-M. Conclusion: Many of the inflammatory changes of ALI that were caused by LPS insult were ameliorated by moxifloxacin treatment.

Direct Action of Genistein on the Hypothalamic Neuronal Circuits in Prepubertal Female Rats : Estrogen Receptor Beta($ER{\beta}$) Pathway? (미성숙한 암컷 흰쥐 시상하부의 신경회로에 미치는 Genistein의 직접 작용 : 에스트로겐 수용체 베타아형 경로?)

  • Heo, Hyun-Jin;Lee, Sung-Ho
    • Development and Reproduction
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    • v.15 no.2
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    • pp.179-185
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    • 2011
  • Some phytoestrogens in soy and red wine, for example, might have beneficiary rather than adverse effects. In particular, dietary soy intake seems to be highly correlated with protection of breast cancer, osteoporosis and cardiovascular disorders. However, questions persist on the potential adverse effects of the main soy constituent genistein (GS) on female reproductive physiology. Previously we found that prepubertal exposure to GS could activate the reproductive system of immature female rats leading to precocious puberty onset, and intracerebroventricularly (ICV) injected GS could directly activate hypothalamic kisspeptin-GnRH neuronal circuits in adult female rats. The present study was performed to examine the hypothalamus-specific GS effects in prepubertal female rats and which subtype of estrogen receptor is mediated in this GS effect. Prepubertal female rats (PND 30) were anaesthetized, treated with single dose of GS (3.4 ${\mu}g$/animal), and sacrificed at 2 hrs post-injection. To determine the transcriptional changes of reproductive hormone-related genes in hypothalamus, total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). ICV infusion of GS significantly lowered the transcriptional activities of mTOR (1:$0.361{\pm}0.058$ AU, p<0.001) but increased that of GAD67 (1:$1.285{\pm}0.099$ AU, p<0.05), which are known to act as an upstream modulator of kisspeptin and GnRH neuronal activities in the hypothalamus, respectively. GS administration enhanced significantly the mRNA levels of KiSS-1(1:$1.458{\pm}0.078$ AU, p<0.001), and exerted no effect on the mRNA level of kisspeptin receptor GPR-54 (1:$1.29{\pm}0.08$ AU). GnRH gene expression was significantly decreased in GS-treated group compared to control group (1:$0.379{\pm}0.196$ AU, p<0.05). There was no difference in the mRNA level of $ER{\alpha}$ in the GS-treated group compare to control group (1:$1.180{\pm}0.390$ AU, Fig. 3A). However, icv infusion of GS significantly increased the transcriptional activities of $ER{\beta}$ (1:$4.209{\pm}0.796$ AU, p<0.01, Fig. 3B). Taken together, the present study indicated that the acute exposure to GS could directly alter the hypothalamic GnRH modulating system in prepubertal female rats. Our study strongly suggested the involvement of $ER{\beta}$ pathway in GS's hypothalamus-specific action, and this idea is consistent with the GS's well-known $ER{\beta}$-mediated protective action in breast cancer.

Tributyltin Increases Adipogenesis and Apoptosis of Rat Testicular Interstitial Cells (Tributyltin에 의한 흰쥐 정소 내 간질세포의 지방세포 유도와 세포자연사 증가)

  • Song, Yeon-Hwa;Jung, Ji-Eun;Lee, Hyun-A;Hong, Ji-Hee;Yang, Hyun-Won
    • Development and Reproduction
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    • v.14 no.4
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    • pp.297-306
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    • 2010
  • Tributyltin (TBT), an endocrine disrupting chemical, has been reported to decrease testicular function by causing apoptosis in the testis, but this mechanism is not fully understood. Thus, in this study we examined whether TBT induces adipogenesis of the Leydig cells to find out the correlation between adipogenesis and apoptosis in the testis. Three week old SD male rats were orally administrated with sesame oil, 1 mg/kg of TBT, or 10 mg/kg of TBT daily for 1 week and weighed after administration. The testes obtained on day 8 were weighed and stained with BODIPY and TUNEL kit. Using total RNA extracted from the isolated Leydig cells, adipogenesis and apoptosis-related genes were analyzed by real-time PCR. The testicular weights of the rats treated with 10 mg/kg TBT were significantly decreased compared to those in the control rats treated with sesame oil. As a result of BODIPY staining, the number of Leydig cells stained with BODIPY was increased in the rats treated with 10 mg/kg TBT compared with the control rats. Similar to BODIPY staining results, the TUNEL assay showed that the apoptosis of Leydig cells was increased in TBT treated rats. The results of the gene expression analysis in the Leydig cells showed that the expression of adipogenesis-related genes (PPAR${\gamma}$, aP2, Perilipin, CD36) and apoptosis-related genes (TNFRSF1A, TNFSF10) was increased after TBT administration. The present study demonstrates that TBT induces the expression of adipogenesis-related and apoptosis-related genes in the Leydig cells leading to adipogenesis and apoptosis in the testes. These results suggest that the dysfunction of Leydig cells by TBT exposure may cause a loss in testicular function.

Histological Changes in Rat Testis by Injection of Hypertonic Saline (고장성 식염수 주사에 의한 흰쥐 정소의 해부학적 변화)

  • Kwak, Byung-Kuk;Lee, Chul-Sang;Lee, Sung-Ho
    • Development and Reproduction
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    • v.14 no.4
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    • pp.281-286
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    • 2010
  • Recent evidence has revealed that the intratesticular injection of hypertonic saline(20%) resulted in a chemically castrated state such as nadir testosterone levels in rats. To confirm the efficacy of this simple saline-injection method further, we investigated the changes in the gross and microscopic anatomy of testis. Our study comprised three groups; intact(control) group, orchidectomy group and saline-injection (experimental) group. Single dose of hypertonic saline (sterilized, $750{\mu}{\ell}/testis$) were directly administered into both testis of adult rats (about 300 g BW). Bilateral orchidectomy was performed at the same day of saline injection. Following 30 days post-injection, reproductive tissues were surgically removed, weighed and fixed for histological examination. The body weights were not changed in both orchidectomy group and saline-injection group when compared to those in intact group. The wet weights of testis were significantly decreased in saline-injection group when compared to those in intact group. The wet weights of epididymis and seminal vesicle and prostate were significantly decreased in orchidectomy group and saline-injection group when compared to those in intact group. Macroscopically, the testes exerted slight atrophy and the tunica albuginea seemed to be intact in saline injection group. Histologically, however, larger parts of testicular tissue underwent necrosis and were barely recognizable after hematoxylin-eosin staining. In the same section, only the opposite part of the injection site was stained showing abnormal state of cell layers mostly fibrosis and infiltrated leukocytes. Sloughing of immature germ cells from the basement membrane along with shedding cells in the intraluminal space was notable in most seminiferous tubules from the saline injected testis. The present study confirmed that the direct injection of hypertonic saline into testis can induce a castration-like, testosterone-depriving effects on accessory sex organs. Our findings suggest that the efficacy of this less expensive and minimally invasive method seems to be almost even with that of conventional orchidectomy and chemical castration, though more in-depth evaluation should be supported.