Superoxide dismutase （SOD） is one of the most important antioxidant defense enzymes, and is considered as the first line against oxidative stress. In this study, we cloned a mitochondrial manganese （Mn） SOD （mMn...Superoxide dismutase （SOD） is one of the most important antioxidant defense enzymes, and is considered as the first line against oxidative stress. In this study, we cloned a mitochondrial manganese （Mn） SOD （mMnSOD） cDNA from the ridgetail white prawn Exopalaemon carinicauda by using rapid amplification of cDNA ends （RACE） methods. The fulMength cDNA for mMnSOD was 1 014-bp long, containing a 5＇-untranslated region （UTR） of 37-bp, a 3＇-UTR of 321-bp with a poly （A） tail, and included a 657-bp open reading frame encoding a protein of 218 amino acids with a 16-amino-acid signal peptide. The protein had a calculated molecular weight of 23.87 kDa and a theoretical isoelectric point of 6.75. The mMnSOD sequence included two putative N-glycosylation sites （NHT and NLS）, the MnSOD signature sequence 18~DVWEHAYY^87, and four putative Mn binding sites （H48, H96, D180, and H184）. Sequence comparison showed that the mMnSOD deduced amino acid sequence of E. carinicauda shared 97%, 95%, 89%, 84%, 82%, 72%, and 69% identity with that ofMacrobrachium rosenbergii, Macrobrachium nipponense, Fenneropeneaus chinensis, Callinectes sapidus, Perisesarma bidens, Danio rerio, and Homo sapiens, resectively. Quantitative real-time RT-PCR analysis showed that mMnSOD transcripts were present in all E. carinicauda tissues examined, with the highest levels in the hepatopancreas. During an ammonia stress treatment, the transcript levels of mMnSOD and cMnSOD were up-regulated at 12 h in hemocytes and at 24 h in the hepatopancreas. As the duration of the ammonia stress treatment extended to 72 h, the transcript levels of mMnSOD and cMnSOD significantly decreased both in hemocytes and hepatopancreas. These findings indicate that the SOD system is induced to respond to acute ammonia stress, and may be involved in environmental stress responses in E. carinicauda.展开更多
A thermostable superoxide dismutase （SOD） from the inshore thermophile Thermus sp. JM1 was purified to homogeneity by steps of fractional ammonium sulfate precipitation, DEAE-Sepharose chromatography and Phenyl-Seph...A thermostable superoxide dismutase （SOD） from the inshore thermophile Thermus sp. JM1 was purified to homogeneity by steps of fractional ammonium sulfate precipitation, DEAE-Sepharose chromatography and Phenyl-Sepharose chromatography. The specific activity of the purified native enzyme was 1 656 U/mg. A sod gene from this strain was cloned and overexpressed in Escherichia coli （E. coli）. The prepared apo-enzyme of the purified recombinant SOD （rSOD） was reconstituted with either Fe or Mn by means of incubation with appropriate metal salts. As a result, only Mn 2＋ - reconstituted rSOD （Mn-rSOD） exhibited the specific activity of 1 598 U/mg. SOD from Thermus sp. JM1 was Mn-SOD, judging by the specific activities analysis of Fe or Mn reconstituted rSODs and the insensitivity of the native SOD to both cyanide and H 2 O 2 . Both the native SOD and Mn- rSOD were determined to be homotetramers with monomeric molecular mass of 26 kDa and 27.5 kDa, respectively. They had high thermostability at 50 ° C and 60 ° C, and showed striking stability across a wide pH span from 4.0 to 11.0.展开更多
In this experiment,the cancer tissues and cells,Which were derived from Lewis lung cancer and A549 lung Cancer cell line,were respectively divided into four groups and zinc, manganese and selenium were respectively ad...In this experiment,the cancer tissues and cells,Which were derived from Lewis lung cancer and A549 lung Cancer cell line,were respectively divided into four groups and zinc, manganese and selenium were respectively added to the medium for 24 hours. The superoxide dismutase activity in the tissues and the cells was estimated. It was found that the SOD activity was enhanced by zinc and manganese and the effect of zinc on SOD activity was superior to that of manganese. We supposed that the enhance of the SOD activity was relative to the activation of the SOD apoenzymes. This experimental result indicated that the inhibitory effect of zinc and manganese on carcinogenesis was achieved by SOD and the elements might be considered a SOD activator.展开更多
AIM:To evaluate the clinical significance of oxidative stress markers in patients with hepatitis C virus(HCV)related hepatocellular carcinoma(HCC).METHODS:Sixty-four consecutive patients who were admitted to Kagoshima...AIM:To evaluate the clinical significance of oxidative stress markers in patients with hepatitis C virus(HCV)related hepatocellular carcinoma(HCC).METHODS:Sixty-four consecutive patients who were admitted to Kagoshima University Medical and Dental Hospital were enrolled in this retrospective study.All patients had chronic liver disease(CLD) due to infection with HCV.Thirty patients with HCV-related HCC,34 with HCV-related CLD without HCC(non-HCC),and 20 healthy volunteers(HVs) were enrolled.Possible associations between serum manganese superoxide dismutase(MnSOD) and thioredoxin(TRX) levels and clinical parameters or patient prognosis were analyzed over a mean follow-up period of 31.7 mo.RESULTS:The serum MnSOD levels were significantly higher in patients with HCV-related HCC than in patients without HCC(P = 0.03) or HVs(P < 0.001).Similarly,serum TRX levels were also significantly higher in patients with HCV-related HCC than in patients without HCC(P = 0.04) or HVs(P < 0.01).However,serum levels of MnSOD and TRX were not correlated in patients with HCC.Among patients with HCC,the overall survival rate(OSR) was lower in patients with MnSOD levels ≥ 110 ng/mL than in patients with levels < 110 ng/mL(P = 0.01),and the OSR tended to be lower in patients with TRX levels < 80 ng/mL(P = 0.05).In addition,patient prognosis with HCC was poorest with serum MnSOD levels ≥ 110 ng/mL and serum TRX levels < 80 ng/mL.Furthermore,a multivariate analysis using a Cox proportional hazard model and serum levels of five factors(MnSOD,prothrombin time,serum albumin,serum α-fetoprotein(AFP),and serum des-γ-carboxy prothrombin) revealed that MnSOD levels ≥ 110 ng/mL(risk ratio:4.12,95% confidential interval:1.22-13.88,P = 0.02) and AFP levels ≥ 40 ng/mL(risk ratio:6.75;95% confidential interval:1.70-26.85,P < 0.01) were independent risk factors associated with a poor patient prognosis.CONCLUSION:Serum MnSOD and TRX levels are potential clinical biomarkers that predict patient prognosis in HCV-related HCC.展开更多
Suppression subtractive hybridization (SSH) was employed to investigate bioluminescence in Panellus stipticus (Bull.) P. Karst. by detecting proteins differentially expressed in bioluminescent and luminescent strains....Suppression subtractive hybridization (SSH) was employed to investigate bioluminescence in Panellus stipticus (Bull.) P. Karst. by detecting proteins differentially expressed in bioluminescent and luminescent strains. Comparisons of luminescent and non-luminescent monokaryon cultures of North American strains revealed differences in transcript levels of proteins responsible for post-translational modification (PTM) of enzymes. A similar comparison of a luminescent strain of P. stipticus from North America with a non-luminescent European strain revealed the presence of extracellular manganese superoxide dismutase (MnSOD) in the luminescent form, in addition to proteins involved in PTM. The application of MnSOD-specific inhibitors to luminescent mycelium resulted in the rapid loss of luminescence. The relevance to luminescence of proteins involved in PTM is discussed, together with a possible role for MnSOD that considers the potential for SODs to form stable complexes with catechols revealed in previously published research. In light of the recent discovery that hispidine may be the precursor of fungal luciferin, we consider a hypothetical mechanism for fungal luminescence in which the ο-hydroquinone moiety of a hispidine derivative ligates with the extracellular form of MnSOD producing a semiquinone-radical complex, with the resultant semiquinonato complex potentially reacting with molecular oxygen or other reactive oxygen species to produce sufficiently excited intermediates to emit light on relaxation.展开更多
目的探讨血清8-羟基脱氧鸟苷酸(8-OHdG)、锰超氧化物歧化酶(MnSOD)水平与急性缺血性脑卒中患者(AIS)经重组组织型纤溶酶原激活剂(rt-PA)静脉溶栓治疗后认知功能恢复情况的相关性。方法选取2016年3月至2018年10月在本院进行rt-PA静脉溶...目的探讨血清8-羟基脱氧鸟苷酸(8-OHdG)、锰超氧化物歧化酶(MnSOD)水平与急性缺血性脑卒中患者(AIS)经重组组织型纤溶酶原激活剂(rt-PA)静脉溶栓治疗后认知功能恢复情况的相关性。方法选取2016年3月至2018年10月在本院进行rt-PA静脉溶栓治疗的112例AIS患者,根据患者溶栓治疗后7 d蒙特利尔认知评估量表(MoCA)评分分为2组:MoCA≥26分的为认知功能恢复良好组(52例),MoCA<26分的为认知功能恢复不良组(60例)。比较两组一般资料及实验室指标的差异,分析可能影响AIS患者rt-PA静脉溶栓后认知功能恢复的因素。结果认知功能恢复良好组与认知功能恢复不良组相比,房纤颤史、美国国立卫生院神经功能缺损(NIHSS)评分、高血脂史、糖尿病史、发病到溶栓时间、8-OHdG、MnSOD、空腹血糖(FPG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)差异均有统计学意义(P<0.05);Pearson相关分析结果显示,认知功能恢复不良组血清8-OHdG与溶栓治疗后7 d MoCA评分呈负相关(P<0.05),MnSOD与溶栓治疗后7 d MoCA评分呈正相关(P<0.05);多因素Logistic回归分析表明,8-OHdG、FPG、TC、LDL-C、发病到溶栓时间均是AIS患者rt-PA静脉溶栓后认知功能恢复的危险因素(P<0.05),MnSOD是保护因素(P<0.05)。结论8-OHdG、MnSOD、房纤颤史、NIHSS评分、高血脂史、糖尿病史、发病到溶栓时间、FPG、TC、LDL-C均与AIS患者rt-PA静脉溶栓后认知功能恢复情况密切相关,可为预测临床预后提供参考依据。展开更多
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2012AA10A409)the Modern AgroIndustry Technology Research System(No.CARS-47)+1 种基金the Special Fund for Independent Innovation of Shandong Province(No.2013CX80202)the Special Fund for Agro-Scientific Research in the Public Interest(No.201103034)
文摘Superoxide dismutase （SOD） is one of the most important antioxidant defense enzymes, and is considered as the first line against oxidative stress. In this study, we cloned a mitochondrial manganese （Mn） SOD （mMnSOD） cDNA from the ridgetail white prawn Exopalaemon carinicauda by using rapid amplification of cDNA ends （RACE） methods. The fulMength cDNA for mMnSOD was 1 014-bp long, containing a 5＇-untranslated region （UTR） of 37-bp, a 3＇-UTR of 321-bp with a poly （A） tail, and included a 657-bp open reading frame encoding a protein of 218 amino acids with a 16-amino-acid signal peptide. The protein had a calculated molecular weight of 23.87 kDa and a theoretical isoelectric point of 6.75. The mMnSOD sequence included two putative N-glycosylation sites （NHT and NLS）, the MnSOD signature sequence 18~DVWEHAYY^87, and four putative Mn binding sites （H48, H96, D180, and H184）. Sequence comparison showed that the mMnSOD deduced amino acid sequence of E. carinicauda shared 97%, 95%, 89%, 84%, 82%, 72%, and 69% identity with that ofMacrobrachium rosenbergii, Macrobrachium nipponense, Fenneropeneaus chinensis, Callinectes sapidus, Perisesarma bidens, Danio rerio, and Homo sapiens, resectively. Quantitative real-time RT-PCR analysis showed that mMnSOD transcripts were present in all E. carinicauda tissues examined, with the highest levels in the hepatopancreas. During an ammonia stress treatment, the transcript levels of mMnSOD and cMnSOD were up-regulated at 12 h in hemocytes and at 24 h in the hepatopancreas. As the duration of the ammonia stress treatment extended to 72 h, the transcript levels of mMnSOD and cMnSOD significantly decreased both in hemocytes and hepatopancreas. These findings indicate that the SOD system is induced to respond to acute ammonia stress, and may be involved in environmental stress responses in E. carinicauda.
基金The Natural Science Foundation of Fujian Province,China under contract Nos 2008J0067 and 2009J01033the Program for New Century Excellent Talents in Fujian Province University under contract No.NCETFJ-2007the Foundation for Innovative Research Team of Jimei University under contract No.2010A005
文摘A thermostable superoxide dismutase （SOD） from the inshore thermophile Thermus sp. JM1 was purified to homogeneity by steps of fractional ammonium sulfate precipitation, DEAE-Sepharose chromatography and Phenyl-Sepharose chromatography. The specific activity of the purified native enzyme was 1 656 U/mg. A sod gene from this strain was cloned and overexpressed in Escherichia coli （E. coli）. The prepared apo-enzyme of the purified recombinant SOD （rSOD） was reconstituted with either Fe or Mn by means of incubation with appropriate metal salts. As a result, only Mn 2＋ - reconstituted rSOD （Mn-rSOD） exhibited the specific activity of 1 598 U/mg. SOD from Thermus sp. JM1 was Mn-SOD, judging by the specific activities analysis of Fe or Mn reconstituted rSODs and the insensitivity of the native SOD to both cyanide and H 2 O 2 . Both the native SOD and Mn- rSOD were determined to be homotetramers with monomeric molecular mass of 26 kDa and 27.5 kDa, respectively. They had high thermostability at 50 ° C and 60 ° C, and showed striking stability across a wide pH span from 4.0 to 11.0.
文摘In this experiment,the cancer tissues and cells,Which were derived from Lewis lung cancer and A549 lung Cancer cell line,were respectively divided into four groups and zinc, manganese and selenium were respectively added to the medium for 24 hours. The superoxide dismutase activity in the tissues and the cells was estimated. It was found that the SOD activity was enhanced by zinc and manganese and the effect of zinc on SOD activity was superior to that of manganese. We supposed that the enhance of the SOD activity was relative to the activation of the SOD apoenzymes. This experimental result indicated that the inhibitory effect of zinc and manganese on carcinogenesis was achieved by SOD and the elements might be considered a SOD activator.
基金Supported by (in part) Grants from the Ministry of Education,Culture, Sports, Science and Technology of Japan, and the Ministry of Health, Labour and Welfare of Japan
文摘AIM:To evaluate the clinical significance of oxidative stress markers in patients with hepatitis C virus(HCV)related hepatocellular carcinoma(HCC).METHODS:Sixty-four consecutive patients who were admitted to Kagoshima University Medical and Dental Hospital were enrolled in this retrospective study.All patients had chronic liver disease(CLD) due to infection with HCV.Thirty patients with HCV-related HCC,34 with HCV-related CLD without HCC(non-HCC),and 20 healthy volunteers(HVs) were enrolled.Possible associations between serum manganese superoxide dismutase(MnSOD) and thioredoxin(TRX) levels and clinical parameters or patient prognosis were analyzed over a mean follow-up period of 31.7 mo.RESULTS:The serum MnSOD levels were significantly higher in patients with HCV-related HCC than in patients without HCC(P = 0.03) or HVs(P < 0.001).Similarly,serum TRX levels were also significantly higher in patients with HCV-related HCC than in patients without HCC(P = 0.04) or HVs(P < 0.01).However,serum levels of MnSOD and TRX were not correlated in patients with HCC.Among patients with HCC,the overall survival rate(OSR) was lower in patients with MnSOD levels ≥ 110 ng/mL than in patients with levels < 110 ng/mL(P = 0.01),and the OSR tended to be lower in patients with TRX levels < 80 ng/mL(P = 0.05).In addition,patient prognosis with HCC was poorest with serum MnSOD levels ≥ 110 ng/mL and serum TRX levels < 80 ng/mL.Furthermore,a multivariate analysis using a Cox proportional hazard model and serum levels of five factors(MnSOD,prothrombin time,serum albumin,serum α-fetoprotein(AFP),and serum des-γ-carboxy prothrombin) revealed that MnSOD levels ≥ 110 ng/mL(risk ratio:4.12,95% confidential interval:1.22-13.88,P = 0.02) and AFP levels ≥ 40 ng/mL(risk ratio:6.75;95% confidential interval:1.70-26.85,P < 0.01) were independent risk factors associated with a poor patient prognosis.CONCLUSION:Serum MnSOD and TRX levels are potential clinical biomarkers that predict patient prognosis in HCV-related HCC.
文摘Suppression subtractive hybridization (SSH) was employed to investigate bioluminescence in Panellus stipticus (Bull.) P. Karst. by detecting proteins differentially expressed in bioluminescent and luminescent strains. Comparisons of luminescent and non-luminescent monokaryon cultures of North American strains revealed differences in transcript levels of proteins responsible for post-translational modification (PTM) of enzymes. A similar comparison of a luminescent strain of P. stipticus from North America with a non-luminescent European strain revealed the presence of extracellular manganese superoxide dismutase (MnSOD) in the luminescent form, in addition to proteins involved in PTM. The application of MnSOD-specific inhibitors to luminescent mycelium resulted in the rapid loss of luminescence. The relevance to luminescence of proteins involved in PTM is discussed, together with a possible role for MnSOD that considers the potential for SODs to form stable complexes with catechols revealed in previously published research. In light of the recent discovery that hispidine may be the precursor of fungal luciferin, we consider a hypothetical mechanism for fungal luminescence in which the ο-hydroquinone moiety of a hispidine derivative ligates with the extracellular form of MnSOD producing a semiquinone-radical complex, with the resultant semiquinonato complex potentially reacting with molecular oxygen or other reactive oxygen species to produce sufficiently excited intermediates to emit light on relaxation.
文摘目的探讨血清8-羟基脱氧鸟苷酸(8-OHdG)、锰超氧化物歧化酶(MnSOD)水平与急性缺血性脑卒中患者(AIS)经重组组织型纤溶酶原激活剂(rt-PA)静脉溶栓治疗后认知功能恢复情况的相关性。方法选取2016年3月至2018年10月在本院进行rt-PA静脉溶栓治疗的112例AIS患者,根据患者溶栓治疗后7 d蒙特利尔认知评估量表(MoCA)评分分为2组:MoCA≥26分的为认知功能恢复良好组(52例),MoCA<26分的为认知功能恢复不良组(60例)。比较两组一般资料及实验室指标的差异,分析可能影响AIS患者rt-PA静脉溶栓后认知功能恢复的因素。结果认知功能恢复良好组与认知功能恢复不良组相比,房纤颤史、美国国立卫生院神经功能缺损(NIHSS)评分、高血脂史、糖尿病史、发病到溶栓时间、8-OHdG、MnSOD、空腹血糖(FPG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)差异均有统计学意义(P<0.05);Pearson相关分析结果显示,认知功能恢复不良组血清8-OHdG与溶栓治疗后7 d MoCA评分呈负相关(P<0.05),MnSOD与溶栓治疗后7 d MoCA评分呈正相关(P<0.05);多因素Logistic回归分析表明,8-OHdG、FPG、TC、LDL-C、发病到溶栓时间均是AIS患者rt-PA静脉溶栓后认知功能恢复的危险因素(P<0.05),MnSOD是保护因素(P<0.05)。结论8-OHdG、MnSOD、房纤颤史、NIHSS评分、高血脂史、糖尿病史、发病到溶栓时间、FPG、TC、LDL-C均与AIS患者rt-PA静脉溶栓后认知功能恢复情况密切相关,可为预测临床预后提供参考依据。
