茶氨酸处理脑缺血后大鼠海马血红素加氧酶-1表达及细胞凋亡的影响研究
|
摘要
目的:探讨茶氨酸对大鼠的脑保护作用是通过上调血红素加氧酶-1(HO-1)的表达来实现,还是通过自身抗氧化应激和细胞凋亡的能力来实现。方法:健康清洁雄性SD大鼠270只,体重180～220 g,采用随机数字表法,将大鼠分为三组(n=90):假手术组(S组),脑缺血再灌注组(IR组),茶氨酸组(TH组),TH组大鼠全脑缺血3 min后尾静脉注射25%茶氨酸溶液4 ml/kg(1000 mg/kg),其余两组分别尾静脉注射生理盐水4 ml/kg。采用四血管阻塞法建立大鼠全脑缺血再灌注模型。分别于再灌注后2,6,12,24,48 h处死,HE染色观察海马形态学变化;IHC染色、实时定量PCR检测HO-1海马表达变化;TUNEL染色、流式细胞仪观察海马细胞凋亡变化。结果:与S组比较,IR组和TH组海马形态学遭到破坏,HO-1表达增加(P<0.05),细胞凋亡上升(P<0.01);与IR组比较,TH组海马形态学得到改善,各个时间点HO-1表达增加得到抑制(P<0.05),细胞凋亡减低(P<0.01)。结论:茶氨酸的脑保护作用不是通过上调HO-1表达来实现的,而是通过自身的抗氧化应激和细胞凋亡能力来实现。
Objective:A discussion about the protective effect of theanine on rat brain is achieved by up regulate the expression of HO-1,or through the way of anti-oxidative stress and apotosis.Methods:270 healthy,clean,male SD rats,weighed from 180 g～220 g,were assigned to three groups randomly. sham operation group(S group),cerebral ischemia reperfusion group(IR group),theanine treated group(TH group). Rats in TH group with intravenous injection of 25% theanine solution 4 ml/kg(1000 mg/kg)when the time of cerebral ischemia reperfusion. The other two groups were with intravenous injection of saline 4 ml/kgrespectively.Four vessels occlusion method was used to establish the model of global cerebral ischemia reperfusion. Rats were put to death after 2 h,6 h,12 h,24 h,48 h of reperfusion,and then observed the histological and cytological changes of hippocampal formation by HE staining,the expression of HO-1 by real-time PCR and observed apoptosis in hippocampal by immunohistochemical staining and TUNEL staining respectively. Results:Compared to S group,the morphology of hippocamal in IR and TH group was destructed,the expression of HO-1 was increased(P<0.05),cell apoptosis was increased(P<0.01);compared to IR group,the morphology of TH group in hippocampal was improved(P<0.05),the increase of HO-1 expression was inhibited(P<0.05),cell apoptosis was decreased(P<0.01). Conclusion:The protective effect of theanin was achieved by its own effect of against oxidative stress and apoptosis,but not through upregulated the expression of HO-1.
Objective:A discussion about the protective effect of theanine on rat brain is achieved by up regulate the expression of HO-1,or through the way of anti-oxidative stress and apotosis.Methods:270 healthy,clean,male SD rats,weighed from 180 g～220 g,were assigned to three groups randomly. sham operation group(S group),cerebral ischemia reperfusion group(IR group),theanine treated group(TH group). Rats in TH group with intravenous injection of 25% theanine solution 4 ml/kg(1000 mg/kg)when the time of cerebral ischemia reperfusion. The other two groups were with intravenous injection of saline 4 ml/kgrespectively.Four vessels occlusion method was used to establish the model of global cerebral ischemia reperfusion. Rats were put to death after 2 h,6 h,12 h,24 h,48 h of reperfusion,and then observed the histological and cytological changes of hippocampal formation by HE staining,the expression of HO-1 by real-time PCR and observed apoptosis in hippocampal by immunohistochemical staining and TUNEL staining respectively. Results:Compared to S group,the morphology of hippocamal in IR and TH group was destructed,the expression of HO-1 was increased(P<0.05),cell apoptosis was increased(P<0.01);compared to IR group,the morphology of TH group in hippocampal was improved(P<0.05),the increase of HO-1 expression was inhibited(P<0.05),cell apoptosis was decreased(P<0.01). Conclusion:The protective effect of theanin was achieved by its own effect of against oxidative stress and apoptosis,but not through upregulated the expression of HO-1.
引文
[1] Mattle HP,Arnold M,Georgiadis D,et al.Comparison of intraarterial and intravenous thrombolysis for ischemic stroke with hyperdense middle cerebral artery sign[J].Stroke,2008,39(2):379-383.
[2] Wang Q,Luo W,Zhou Q.Intervention of NGAL and HO-1 in valve replacement surgery-induced acute kidney injury[J].Journal of Central South University,2014,39(10):1001.
[3] Yang C,Zhang X,Fan H,et al.Curcumin upregulates transcription factor Nrf2,HO-1 expression and protects rat brains against focal ischemia[J].Brain Research,2009,1282(12):133-141.
[4] 鲍昕,王宁,李传坤,等.大鼠脑出血后血肿周围组织中p75NTR、TrkA的表达及其与细胞凋亡的关系[J].陕西医学杂志,2014,43(9):1112-1115.
[5] Lv W.Relationship between neural apoptosis and cell death inducing Dff45-like effecter-B gene expression after cerebral ischemic reperfusion injury in rats[J].Journal of the Neurological Sciences,2013,333(4):e153-e153.
[6] 王宁,张珍妮,吕建瑞,等.茶氨酸预先给药对大鼠脑缺血再灌注损伤时神经元DNA修复功能的影响[J].中华麻醉学杂志,2016,36(4):414-417.
[7] Zheng JM,Chen XC,Lin M,et al.Mechanism of the reduction of cerebral ischemic-reperfusion injury through inhibiting the activity of NF-kappaB by propyl gallate[J].Acta Pharmaceutica Sinica,2011,46(2):158.
[8] Kakuda T,Nozawa A,Kubo T,et al.Neuroprotection of glutamylathylamide(theanine) against kainic acid toxicity in rat hippocampus[J].Society For Neuroscience,2001,334:15-15.
[9] 吕飒美,吴友伟,薛挥.诱导血红素氧合酶-1对大鼠肝脏缺血再灌注的影响[J].陕西医学杂志,2010,39(2):150-152.
[10] 郭筱华,赵忠新.血红素加氧酶系统在神经系统疾病中变化的研究进展[J].中国临床神经科学,2007,15(2):212-216.
[11] Connell BJ,Saleh MC,Khan BV,et al.Apocynin may limit total cell death following cerebral ischemia and reperfusion by enhancing apoptosis[J].Food & Chemical Toxicology,2011,49(12):3063-3069.
[2] Wang Q,Luo W,Zhou Q.Intervention of NGAL and HO-1 in valve replacement surgery-induced acute kidney injury[J].Journal of Central South University,2014,39(10):1001.
[3] Yang C,Zhang X,Fan H,et al.Curcumin upregulates transcription factor Nrf2,HO-1 expression and protects rat brains against focal ischemia[J].Brain Research,2009,1282(12):133-141.
[4] 鲍昕,王宁,李传坤,等.大鼠脑出血后血肿周围组织中p75NTR、TrkA的表达及其与细胞凋亡的关系[J].陕西医学杂志,2014,43(9):1112-1115.
[5] Lv W.Relationship between neural apoptosis and cell death inducing Dff45-like effecter-B gene expression after cerebral ischemic reperfusion injury in rats[J].Journal of the Neurological Sciences,2013,333(4):e153-e153.
[6] 王宁,张珍妮,吕建瑞,等.茶氨酸预先给药对大鼠脑缺血再灌注损伤时神经元DNA修复功能的影响[J].中华麻醉学杂志,2016,36(4):414-417.
[7] Zheng JM,Chen XC,Lin M,et al.Mechanism of the reduction of cerebral ischemic-reperfusion injury through inhibiting the activity of NF-kappaB by propyl gallate[J].Acta Pharmaceutica Sinica,2011,46(2):158.
[8] Kakuda T,Nozawa A,Kubo T,et al.Neuroprotection of glutamylathylamide(theanine) against kainic acid toxicity in rat hippocampus[J].Society For Neuroscience,2001,334:15-15.
[9] 吕飒美,吴友伟,薛挥.诱导血红素氧合酶-1对大鼠肝脏缺血再灌注的影响[J].陕西医学杂志,2010,39(2):150-152.
[10] 郭筱华,赵忠新.血红素加氧酶系统在神经系统疾病中变化的研究进展[J].中国临床神经科学,2007,15(2):212-216.
[11] Connell BJ,Saleh MC,Khan BV,et al.Apocynin may limit total cell death following cerebral ischemia and reperfusion by enhancing apoptosis[J].Food & Chemical Toxicology,2011,49(12):3063-3069.