重组草酸降解酶乳酸菌治疗高草酸尿症的实验研究

发布时间：2018-06-08 22:35

本文选题：草酸脱羧酶 + 草酸氧化酶　；参考：《华中科技大学学报(医学版)》2017年05期

【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。
[Abstract]:Objective to construct lactic acid bacteria expressing recombinant oxalic acid degrading enzyme and to investigate the therapeutic effect of lactobacillus on rats with hyperoxaliduria. Methods the oxalate decarboxylaseODCand oxalate oxidase Oxo were recombined with the expression plasmid pMG36e, and the recombinant plasmid was transformed into the lactobacillus MG1363 by electroporation. The culture medium containing 100 mmol / L oxalic acid for 48 h was used to measure the growth and the content of oxalic acid in the medium. SD rats were fed with 5% oxalic acid and were treated with recombinant lactic acid bacteria for 1 week. The urine of rats was collected for 24 hours to measure oxalic acid content. Results Recombinant pMG36e-ODCG pMG36e-OxO lactic acid bacteria were successfully constructed. Both of them could grow in the medium containing 100 mmol / L oxalic acid for about 20 h and enter the logarithmic phase about 40 h into stable phase. Among them, pMG36e-ODC lactic acid bacteria could significantly reduce oxalic acid concentration in culture medium and degrade 30% within 48 h, while the oxalic acid content in culture medium did not change significantly after culture of pMG36e-OxO lactobacillus and normal lactic acid bacteria. Within 48 hours, oxalic acid content decreased about 15.pMG36eODC lactobacillus. It can significantly reduce the content of oxalic acid in urine of rats with high oxalic acid diet. Conclusion Recombinant ODC lactic acid bacteria can effectively degrade oxalic acid and alleviate hyperoxalic acid urination in model rats by degrading oxalic acid in food.
【作者单位】：华中科技大学同济医学院附属同济医院泌尿外科;湖北大学生命科学学院;武汉市普爱医院泌尿外科;
【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)
【分类号】：R691.4

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