目的:肠道作为人体的重要消化器官,其内定植的微生物在尿酸合成和代谢过程中发挥着重要作用,本研究利用含尿酸靶向培养基筛选正常人群肠道内具有降尿酸功能的细菌并鉴定。方法:依据尿酸的摩尔质量制备含不同浓度尿酸的BHI培养基,液体培...目的:肠道作为人体的重要消化器官,其内定植的微生物在尿酸合成和代谢过程中发挥着重要作用,本研究利用含尿酸靶向培养基筛选正常人群肠道内具有降尿酸功能的细菌并鉴定。方法:依据尿酸的摩尔质量制备含不同浓度尿酸的BHI培养基,液体培养基扩增并驯化肠道粪便微生物,固体培养基分离和纯化具有尿酸降解功能的细菌。挑取固体培养基上形态一致的单个菌落进行革兰氏染色和镜检,筛选出已纯化菌株,在需氧和厌氧培养条件下测定尿酸降解率,选取降解率≥50%以上的菌株为高效尿酸降解菌的候选菌株,再测定不同温度和pH值条件下的尿酸降解率,进行降尿酸条件优化。利用16S r DNA序列测定法对尿酸降解菌进行鉴定,药敏实验测定该菌对抗生素的敏感性。结果:正常人群粪便微生物中分离获得一株高效尿酸降解菌B5C,第5天需氧条件下的尿酸降解率均>50%,与初始尿酸浓度相比具有统计学意义(P<0.05)。优化降尿酸条件后,在37℃、pH7.0时,降解率可达88.7%,经鉴定为粪肠球菌,对常见的抗生素如阿莫西林、氨苄西林和青霉素G等具有较高的敏感性。结论:本研究利用含不同尿酸浓度的靶向培养基驯化、分离和鉴定出一株人肠源性细菌,在需氧条件下也具有较高的尿酸降解率,可为今后临床降尿酸微生物制剂的开发和利用提供新的菌种资源。展开更多
Objective：To observe the effect of acupuncture on the expression of mitochondrial proteome in hippocampus of senescence-accelerated mouse prone g （SAMPg） mice models with Alzheimer disease （AD）,and to explore the...Objective：To observe the effect of acupuncture on the expression of mitochondrial proteome in hippocampus of senescence-accelerated mouse prone g （SAMPg） mice models with Alzheimer disease （AD）,and to explore the possible protective mechanism of acupuncture on mitochondria.Methods：Sixty 6-month-old male SAMP8 mice were randomly divided into an acupuncture at acupoint group,an acupuncture at non-acupoint group and a model group,20 mice in each group.The 20 male senescence-accelerated mouse/resistance 1 （SAMR1） mice of the same age were used as a normal control group.Shenshu （BL 23）,Baihui （GV 20）,Xuehai （SP 10） and Geshu （BL 17） were selected for acupuncture intervention in acupuncture at acupoint group.After an 8-week intervention,mitochondrial tissues were extracted from the hippocampus.Differentially expressed proteins were identified by subcellular organelle proteomics.Western blot was used to verify the expressions of some related proteins in hippocampal mitochondria.Results：Compared with the model group,there were 13 differentially expressed protein spots in the acupuncture at acupoint group,of which,9 were up-regulated,including neurofilament light polypeptide （NFL）,actin （cytoplasmic 1,database ID：ACTB）,tubulin beta-2A chain （TBB2A）,tropomodulin-2 （TMOD2）,pyruvate dehydrogenase E1 component subunit beta （PDHE1-β）,NADH-ubiquinone oxidoreductase 75 kDa subunit （database ID：NDUS1）,heat shock cognate 71 kDa protein （HSC71）,pyruvate dehydrogenase E1 component subunit alpha （PDHE1-α） and ATP synthase beta subunit （ATP-β）;4 were down-regulated,including glial fibrillary acidic protein （GFAP）,pyruvate dehydrogenase phosphatase 1 （PDP1）,mitochondrial-processing peptidase subunit alpha （MMP-α） and adenosine kinase （ADK）.According to the information provided in the protein database,most of the differentially expressed proteins involve the regulation of mitochondrial function and structure.The expression levels of NFL and TBB2A in the normal control group and the acupuncture at acupoint group were significantly higher than those in the acupuncture at non-acupoint group （P〈0.05）.ATP-β and NDUS1 expression levels were significantly higher in the acupuncture at acupoint group than those in the acupuncture at non-acupoint group （P〈0.05）;there was no significant difference between the acupuncture at non-acupoint group and the model group （P〉0.05）.Conclusion：Acupuncture may achieve the potential therapeutic effect on AD by regulating the structure and functional proteins of hippocampal mitochondria.展开更多
文摘目的:肠道作为人体的重要消化器官,其内定植的微生物在尿酸合成和代谢过程中发挥着重要作用,本研究利用含尿酸靶向培养基筛选正常人群肠道内具有降尿酸功能的细菌并鉴定。方法:依据尿酸的摩尔质量制备含不同浓度尿酸的BHI培养基,液体培养基扩增并驯化肠道粪便微生物,固体培养基分离和纯化具有尿酸降解功能的细菌。挑取固体培养基上形态一致的单个菌落进行革兰氏染色和镜检,筛选出已纯化菌株,在需氧和厌氧培养条件下测定尿酸降解率,选取降解率≥50%以上的菌株为高效尿酸降解菌的候选菌株,再测定不同温度和pH值条件下的尿酸降解率,进行降尿酸条件优化。利用16S r DNA序列测定法对尿酸降解菌进行鉴定,药敏实验测定该菌对抗生素的敏感性。结果:正常人群粪便微生物中分离获得一株高效尿酸降解菌B5C,第5天需氧条件下的尿酸降解率均>50%,与初始尿酸浓度相比具有统计学意义(P<0.05)。优化降尿酸条件后,在37℃、pH7.0时,降解率可达88.7%,经鉴定为粪肠球菌,对常见的抗生素如阿莫西林、氨苄西林和青霉素G等具有较高的敏感性。结论:本研究利用含不同尿酸浓度的靶向培养基驯化、分离和鉴定出一株人肠源性细菌,在需氧条件下也具有较高的尿酸降解率,可为今后临床降尿酸微生物制剂的开发和利用提供新的菌种资源。
文摘Objective：To observe the effect of acupuncture on the expression of mitochondrial proteome in hippocampus of senescence-accelerated mouse prone g （SAMPg） mice models with Alzheimer disease （AD）,and to explore the possible protective mechanism of acupuncture on mitochondria.Methods：Sixty 6-month-old male SAMP8 mice were randomly divided into an acupuncture at acupoint group,an acupuncture at non-acupoint group and a model group,20 mice in each group.The 20 male senescence-accelerated mouse/resistance 1 （SAMR1） mice of the same age were used as a normal control group.Shenshu （BL 23）,Baihui （GV 20）,Xuehai （SP 10） and Geshu （BL 17） were selected for acupuncture intervention in acupuncture at acupoint group.After an 8-week intervention,mitochondrial tissues were extracted from the hippocampus.Differentially expressed proteins were identified by subcellular organelle proteomics.Western blot was used to verify the expressions of some related proteins in hippocampal mitochondria.Results：Compared with the model group,there were 13 differentially expressed protein spots in the acupuncture at acupoint group,of which,9 were up-regulated,including neurofilament light polypeptide （NFL）,actin （cytoplasmic 1,database ID：ACTB）,tubulin beta-2A chain （TBB2A）,tropomodulin-2 （TMOD2）,pyruvate dehydrogenase E1 component subunit beta （PDHE1-β）,NADH-ubiquinone oxidoreductase 75 kDa subunit （database ID：NDUS1）,heat shock cognate 71 kDa protein （HSC71）,pyruvate dehydrogenase E1 component subunit alpha （PDHE1-α） and ATP synthase beta subunit （ATP-β）;4 were down-regulated,including glial fibrillary acidic protein （GFAP）,pyruvate dehydrogenase phosphatase 1 （PDP1）,mitochondrial-processing peptidase subunit alpha （MMP-α） and adenosine kinase （ADK）.According to the information provided in the protein database,most of the differentially expressed proteins involve the regulation of mitochondrial function and structure.The expression levels of NFL and TBB2A in the normal control group and the acupuncture at acupoint group were significantly higher than those in the acupuncture at non-acupoint group （P〈0.05）.ATP-β and NDUS1 expression levels were significantly higher in the acupuncture at acupoint group than those in the acupuncture at non-acupoint group （P〈0.05）;there was no significant difference between the acupuncture at non-acupoint group and the model group （P〉0.05）.Conclusion：Acupuncture may achieve the potential therapeutic effect on AD by regulating the structure and functional proteins of hippocampal mitochondria.
