The relationship between myocardial cell damages and HSPs mRNA transcription in heat stressed broilers was studied using a spectrophotometer, the histopathological technique, and fluorescence quantitative reverse tran...The relationship between myocardial cell damages and HSPs mRNA transcription in heat stressed broilers was studied using a spectrophotometer, the histopathological technique, and fluorescence quantitative reverse transcription PCR （FQ RT-PCR）. The results showed that the activities of creatine kinase （CK） and glutamic-pyruvic transaminase （GPT） were induction during the persistent heat stress. The major lesions of the myocardial fibers were granular degeneration and necrosis. The transcription of constitutive or cognate heat shock protein 70 （HSC70） mRNA was changeable. The transcription of heat shock protein 70 （HSPT0） mRNA was increased obviously in the course of persistent heat stress. The results showed that the change of HSC70 mRNA transcription was contrary to the activity of CK, and the level of HSC70 mRNA transcription must be used as a symbol of the myocardial cell damages in the course of persistent heat stress.展开更多
Objective To investigate and compare the effect of radio-frequency(RF)field exposure on expression of heat shock proteins(Hsps)in three human glioma cell lines(MO54,A172,and T98).Methods Cells were exposed to sham or ...Objective To investigate and compare the effect of radio-frequency(RF)field exposure on expression of heat shock proteins(Hsps)in three human glioma cell lines(MO54,A172,and T98).Methods Cells were exposed to sham or 1950 MHz continuous-wave for 1 h.Specific absorption rates(SARs)were 1 and 10 W/kg.Localization and expression of Hsp27 and phosphorylated Hsp27((78)Ser)(p-Hsp27)were examined by immunocytochemistry.Expression levels of Hsp27,p-Hs27,and Hsp70 were determined by Western blotting.Results The Hsp27 was primarily located within the cytoplasm,p-Hsp27 in both cytoplasm and nuclei of MO54,A172,and T98 cells.RF field exposure did not affect the distribution or expression of Hsp27.In addition,Western blotting showed no significant differences in protein expression of Hsp27 or HspT0 between sham-and RF field-exposed cells at a SAR of 1 W/kg and 10 W/kg for 1 h in three cells lines.Exposure to RF field at a SAR of 10 W/kg for 1 h slightly decreased the protein level of phosphorylated Hsp27 in MO54 cells.Conclusion The 1950 MHz RF field has only little or no apparent effect on Hsp70 and Hsp27 expression in MO54,A172,and T98 cells.展开更多
【目的】探究黑龙江林蛙(Rana amurensis)热休克蛋白90α家族B类成员1(heat shock protein 90 alpha class B member 1,HSP90AB1)基因的序列特征,了解其组织表达特性和脂多糖(LPS)诱导下的表达变化。【方法】以黑龙江林蛙脾脏组织cDNA...【目的】探究黑龙江林蛙(Rana amurensis)热休克蛋白90α家族B类成员1(heat shock protein 90 alpha class B member 1,HSP90AB1)基因的序列特征,了解其组织表达特性和脂多糖(LPS)诱导下的表达变化。【方法】以黑龙江林蛙脾脏组织cDNA为模板,通过PCR扩增HSP90AB1基因完整编码区(CDS)序列,使用在线软件对其编码蛋白进行生物信息学分析;利用实时荧光定量PCR和Western blotting检测生理状态下HSP90AB1基因在黑龙江林蛙皮肤、肌肉、心脏、肝脏、脾脏、肺脏、肾脏和胃等组织器官中的表达量,同时对LPS诱导下不同时间点HSP90AB1基因在不同组织器官的转录水平进行检测。【结果】黑龙江林蛙HSP90AB1基因CDS长2178 bp,共编码725个氨基酸。生物信息学分析显示,HSP90AB1蛋白不存在信号肽且无跨膜信号,属于非跨膜蛋白,存在62个潜在磷酸化位点(含33个丝氨酸、19个苏氨酸及10个酪氨酸位点),其二级结构以α-螺旋为主(占47.03%),其次为无规则卷曲(占39.59%)和延伸链(占13.38%)。系统进化树结果显示,黑龙江林蛙与欧洲林蛙亲缘关系最近。在生理状态下,HSP90AB1基因在黑龙江林蛙胃中表达量最高,且显著高于其他组织(P<0.05);在肌肉组织中表达量最低,且显著低于其他组织(P<0.05)。Western blotting结果显示,HSP90AB1蛋白在黑龙江林蛙胃中表达量最高;在心脏中表达量最低,且显著低于其他组织(P<0.05)。LPS诱导后,黑龙江林蛙各组织中HSP90AB1基因表达量随诱导时间延长而呈降低趋势,其中诱导72和96 h表达量均显著低于其他时间点(P<0.05);诱导12、72 h时脾脏中HSP90AB1基因表达量显著高于其他组织(P<0.05);24 h时肌肉和脾脏中HSP90AB1基因表达量显著高于其他组织(P<0.05)。【结论】本研究成功克隆了黑龙江林蛙HSP90AB1基因CDS区序列,并明确了其在不同组织中的表达规律,其中在胃中表达量最高,肌肉组织中表达量最低。LPS诱导可明显抑制HSP90AB1基因在黑龙江林蛙不同组织中的转录水平。本试验结果为深入探究HSP90AB1基因在林蛙生理及应激过程中的功能机制提供了理论依据。展开更多
基金This work was financially supported by the National Natural Science Foundation of China （30170682, 30571400） the Specialized Research Fund for the Doctoral Program of Higher Education, China （20050307008）.
文摘The relationship between myocardial cell damages and HSPs mRNA transcription in heat stressed broilers was studied using a spectrophotometer, the histopathological technique, and fluorescence quantitative reverse transcription PCR （FQ RT-PCR）. The results showed that the activities of creatine kinase （CK） and glutamic-pyruvic transaminase （GPT） were induction during the persistent heat stress. The major lesions of the myocardial fibers were granular degeneration and necrosis. The transcription of constitutive or cognate heat shock protein 70 （HSC70） mRNA was changeable. The transcription of heat shock protein 70 （HSPT0） mRNA was increased obviously in the course of persistent heat stress. The results showed that the change of HSC70 mRNA transcription was contrary to the activity of CK, and the level of HSC70 mRNA transcription must be used as a symbol of the myocardial cell damages in the course of persistent heat stress.
文摘Objective To investigate and compare the effect of radio-frequency(RF)field exposure on expression of heat shock proteins(Hsps)in three human glioma cell lines(MO54,A172,and T98).Methods Cells were exposed to sham or 1950 MHz continuous-wave for 1 h.Specific absorption rates(SARs)were 1 and 10 W/kg.Localization and expression of Hsp27 and phosphorylated Hsp27((78)Ser)(p-Hsp27)were examined by immunocytochemistry.Expression levels of Hsp27,p-Hs27,and Hsp70 were determined by Western blotting.Results The Hsp27 was primarily located within the cytoplasm,p-Hsp27 in both cytoplasm and nuclei of MO54,A172,and T98 cells.RF field exposure did not affect the distribution or expression of Hsp27.In addition,Western blotting showed no significant differences in protein expression of Hsp27 or HspT0 between sham-and RF field-exposed cells at a SAR of 1 W/kg and 10 W/kg for 1 h in three cells lines.Exposure to RF field at a SAR of 10 W/kg for 1 h slightly decreased the protein level of phosphorylated Hsp27 in MO54 cells.Conclusion The 1950 MHz RF field has only little or no apparent effect on Hsp70 and Hsp27 expression in MO54,A172,and T98 cells.
文摘【目的】探究黑龙江林蛙(Rana amurensis)热休克蛋白90α家族B类成员1(heat shock protein 90 alpha class B member 1,HSP90AB1)基因的序列特征,了解其组织表达特性和脂多糖(LPS)诱导下的表达变化。【方法】以黑龙江林蛙脾脏组织cDNA为模板,通过PCR扩增HSP90AB1基因完整编码区(CDS)序列,使用在线软件对其编码蛋白进行生物信息学分析;利用实时荧光定量PCR和Western blotting检测生理状态下HSP90AB1基因在黑龙江林蛙皮肤、肌肉、心脏、肝脏、脾脏、肺脏、肾脏和胃等组织器官中的表达量,同时对LPS诱导下不同时间点HSP90AB1基因在不同组织器官的转录水平进行检测。【结果】黑龙江林蛙HSP90AB1基因CDS长2178 bp,共编码725个氨基酸。生物信息学分析显示,HSP90AB1蛋白不存在信号肽且无跨膜信号,属于非跨膜蛋白,存在62个潜在磷酸化位点(含33个丝氨酸、19个苏氨酸及10个酪氨酸位点),其二级结构以α-螺旋为主(占47.03%),其次为无规则卷曲(占39.59%)和延伸链(占13.38%)。系统进化树结果显示,黑龙江林蛙与欧洲林蛙亲缘关系最近。在生理状态下,HSP90AB1基因在黑龙江林蛙胃中表达量最高,且显著高于其他组织(P<0.05);在肌肉组织中表达量最低,且显著低于其他组织(P<0.05)。Western blotting结果显示,HSP90AB1蛋白在黑龙江林蛙胃中表达量最高;在心脏中表达量最低,且显著低于其他组织(P<0.05)。LPS诱导后,黑龙江林蛙各组织中HSP90AB1基因表达量随诱导时间延长而呈降低趋势,其中诱导72和96 h表达量均显著低于其他时间点(P<0.05);诱导12、72 h时脾脏中HSP90AB1基因表达量显著高于其他组织(P<0.05);24 h时肌肉和脾脏中HSP90AB1基因表达量显著高于其他组织(P<0.05)。【结论】本研究成功克隆了黑龙江林蛙HSP90AB1基因CDS区序列,并明确了其在不同组织中的表达规律,其中在胃中表达量最高,肌肉组织中表达量最低。LPS诱导可明显抑制HSP90AB1基因在黑龙江林蛙不同组织中的转录水平。本试验结果为深入探究HSP90AB1基因在林蛙生理及应激过程中的功能机制提供了理论依据。
