Artesunate interacts with Vitamin D receptor to reverse mouse model of sepsis-induced immunosuppression via enhancing autophagy

Shenglan Shang; Jiaqi Wu; Xiaoli Li; Xin Liu; Pan Li; Chunli Zheng; Yonghua Wang; Songqing Liu; Jiang Zheng; Hong Zhou

Pseudomonas aeruginosa (PA) clinical insolate was kindly provided by Prof. Peiyuan Xia (Southwestern Hospital, Chongqing, China). Single colony was picked from viable, growing Mueller-Hinton (MH) agar plate. Then, it was transferred to 5 ml of liquid MH medium and cultivated aerobically at 37 ℃ in orbital shaking incubator for 4 h. The cultures were transferred to 100 ml of fresh MH medium for another 12 h. At an OD600 of 0.6 -0.8, when the bacterial culture was within the logarithmic phase of growth measured by SmartSpec 3000 spectrophotometer (Bio-Rad, Hercules, CA, USA), the suspension was centrifuged at 1500 g for 10 min to harvest the pellet. After washing twice, the pellet was re-suspended by sterile normal saline. The bacteria were diluted in sterile normal saline to achieve the terminal concentration [(colony-formation units (CFU)/mL], which was calculated according to the bacterial growth curve determined by our lab (Concentration = (9.7544 × OD -1.4965) × 10 8 . CC-BY-NC-ND 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.02.26.966143 doi: bioRxiv preprint CFU/mL).