重组草酸降解酶乳酸菌治疗高草酸尿症的实验研究  

Treatment of Hyperoxaluria with Recombinant Lactobacillus pMG36e-ODC

作　　者：杨欢[1] 赵晨明 朱晓敬 李洋 郑涛[3] 管维[1] 郭小林[1] 陈志强[1] 叶章群[1] Yang Huan Zhao Chenming Zhu Xiaojing et al(Department of Urology, Tongji Hospital, Tongjiji Medical College, Huazhong University of Science and Technology ,Wuhan 430030 ,China College of Life Science, Hubei University ,Wuhan 430063 ,China)

机构地区：[1]华中科技大学同济医学院附属同济医院泌尿外科,武汉430030 [2]湖北大学生命科学学院,武汉430063 [3]武汉市普爱医院泌尿外科,武汉430033

出　　处：《华中科技大学学报（医学版）》2017年第5期563-566,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.81270788;No.81500534;No.81370805);湖北省自然科学基金资助项目(No.2015CFB289;No.2012FFB02412;No.2013CFB174);武汉市青年科技晨光计划项目(No.2013072304010833;No.2015070404010199);教育部高等学校博士学科点专项科研基金新教师基金(No.20130142120072)

摘　　要：目的构建表达重组草酸降解酶的乳酸菌,并探讨对高草酸尿症大鼠的治疗作用。方法将草酸脱羧酶(oxalate decarboxylase,ODC)和草酸氧化酶(oxalate oxidase,OxO)基因与表达质粒pMG36e重组,通过电转化将重组质粒转入乳酸菌MG1363中。在含100mmol/L草酸的培养液中培养48h,测量其生长情况及培养液中草酸含量变化。以含5%草酸食物喂养SD大鼠,并同时进行重组乳酸菌灌胃治疗,1周后收集大鼠24h尿液测量草酸含量。结果成功构建重组pMG36e-ODC,pMG36e-OxO乳酸菌,两者都可在含100mmol/L草酸的培养液中生长,20h左右开始进入对数期,40h左右进入稳定期。其中pMG36e-ODC乳酸菌可明显降低培养液内草酸浓度,48h内可降解30%,而培养pMG36e-OxO乳酸菌和正常乳酸菌后培养液中草酸含量无明显变化,48h内草酸含量水平下降了约15%。pMG36eODC乳酸菌喂养可显著降低高草酸饮食大鼠尿液中草酸含量。结论重组ODC乳酸菌可有效降解草酸,并可通过降解食物中草酸缓解模型大鼠高草酸尿症。Objective To transfer oxalate decarboxylase（ODC）and oxalate oxidase（OxO）into Lactobacillus（Lab）to express oxalate-degrading enzyme,which could continuously degrade oxalate,degrade dietary oxalate via oral administration and prevent the formation of calcium oxalate stones.Methods Recombinant plasmids pMG36 eexpressing ODC or OxO were established and electrotransformed into Labs MG1363,which were cultured in the medium containing 100mmol/L oxalate for 48 h to examine the oxalate degradation.SD rats were fed on 5% oxalate diet and treated with recombinant Labs by gavage.One week later,the 24-h urinary oxalate content was measured.Results The recombinant Labs pMG36e-ODC and pMG36e-OxO were successfully constructed and both could grow in oxalate-containing medium.The growth curve showed that the logarithmic phase began at about 20 hand the stable phase at about 40 h.The recombinant Lab pMG36e-ODC could degrade oxalate significantly,by 30% in 48 h,while the recombinant Lab pMG36e-OxO and the wild type Lab showed little efficiency,and they decreased oxalate by nearly 15% in 48 h.Additionally,the recombinant Lab pMG36e-ODC could significantly decrease the amount of urinary oxalate in rats on high oxalate diet.Conclusion The recombinant Lab pMG36e-ODC degrades oxalate effectively and prevents hyperoxaluria by reducing dietary oxalate.

关 键 词：草酸脱羧酶 草酸氧化酶 乳酸菌 基因重组 草酸钙结石 

分 类 号：R692.4[医药卫生—泌尿科学]