不同培养基中铜绿假单胞菌对秀丽隐杆线虫生长及cyp35家族基因表达的影响

Effects of Pseudomonas aeruginosa in different culture media on the growth and cyp35 family genes expressions of Caenorhabditis elegans

目的研究在不同培养基中生长的铜绿假单胞菌对模式生物秀丽隐杆线虫发育和生殖的影响,观察线虫cyp35家族药物代谢基因表达的变化,探讨铜绿假单胞菌对线虫的毒性作用及代谢机制。方法建立铜绿假单胞菌-秀丽隐杆线虫细菌感染模型,通过LB、TSA、PGS和PGA等培养基培养铜绿假单胞菌感染秀丽隐杆线虫后,观察线虫寿命、形态、产卵和运动等情况;通过实时荧光定量PCR检测铜绿假单胞菌感染秀丽隐杆线虫cyp35家族基因的表达。结果铜绿假单胞菌感染秀丽隐杆线虫后,秀丽隐杆线虫生长发育缓慢,运动的灵活性降低,各铜绿假单胞菌培养基实验组的平均生存时间和产卵量均低于NGM培养基对照组(P<0.05);秀丽隐杆线虫感染铜绿假单胞菌后体内的cyp35A1、cyp35A2、cyp35A5和cyp35C1等毒物-药物代谢基因与对照组比较表达均上调(P<0.05)。结论不同培养基上生长的铜绿假单胞菌对秀丽隐杆线虫有不同的毒性效应;秀丽隐杆线虫对铜绿假单胞菌毒素的代谢与cyp35家族基因密切相关。

Objective To explore the toxic effect and metabolic mechanism of Pseudomonas aeruginosa on edlworm by investigating the effect of Pseudomonas aeruginosa in different culture media on the development and reproduction of Caenorhabditis elegans（ C. elegans）,and observing the changes of cyp35 family genes expressions. Methods The C. elegans model infected by Pseudomonas aeruginosa was established. Then,the C. elegans were cultured in LB,TSA,PGS and PGA media,respectively,and its lifespan,appearance,egg-laying and movement were observed. The expressions of cyp35 family genes including cyp35A1,cyp35A2,cyp35A5 and cyp35C1 were detected by RT-PCR. Results The C. elegans infected by Pseudomonas aeruginosa showed retardant growth and development,and slow movement. The average survival time and egg-laying number of C. elegans in LB,TSA,PGS and PGA media were significantly lower than that in nematode growth medium（ NGM）（ P〈0. 05）. The expression levels of cyp35A1,cyp35A2,cyp35A5 and cyp35C1 in the C. elegans infected by Pseudomonas aeruginosa increased significantly（ P〈0. 05）. Conclusion Pseudomonas aeruginosa in different culture media has different toxic effects on C. elegans,and the toxin metabolism in C. elegans is closely related to cyp35 family genes.