key: cord-0697244-yz01c862 authors: Liu, Xingyun; Xu, Jun; Chang, Xinyu; Chen, Lu; Shen, Ruiting; Ding, Ning; Hou, Xiaolin title: Lithium chloride inhibits infectious bronchitis virus-induced apoptosis and inflammation date: 2021-12-07 journal: Microb Pathog DOI: 10.1016/j.micpath.2021.105352 sha: 92978e17ea571f12be1666c3d95819ed38d14f21 doc_id: 697244 cord_uid: yz01c862 Avian infectious bronchitis (IB) was caused by infectious bronchitis virus (IBV), a coronavirus, which leads to enormous economic losses in the poultry industry. Studies have shown that lithium chloride (LiCl) is a good virus inhibitor. Through cell culture, virus infection, and RT-qPCR, we found that LiCl could down-regulate the apoptosis-related genes Caspase-3 and Bax, up-regulate Bcl-2, and down-regulate the inflammatory-related genes (NF-κB, NLRP3, TNF-α, and IL-1β) via inhibiting virus replication. Finally, clinical trials showed that LiCl could inhibit IBV-induced apoptosis and inflammatory in chicken embryos as well as reduce the mortality and deformity rate of chicken embryos. The results showed that LiCl has antiviral activity against IBV and clinical effects. Further studies are required to explore the exact action mechanism of LiCl on IBV-induced apoptosis and inflammation. coronavirus, which leads to enormous economic losses in the poultry industry. Studies have shown 24 that lithium chloride (LiCl) is a good virus inhibitor. Through cell culture, virus infection, and 25 RT-qPCR, we found that LiCl could down-regulate the apoptosis-related genes Caspase-3 and Bax, 26 up-regulate Bcl-2, and down-regulate the inflammatory-related genes (NF-κB, NLRP3, TNF-α, 27 and IL-1β) via inhibiting virus replication. Finally, clinical trials showed that LiCl could inhibit 28 IBV-induced apoptosis and inflammatory in chicken embryos as well as reduce the mortality and 29 deformity rate of chicken embryos. The results showed that LiCl has antiviral activity against IBV 30 and clinical effects. Further studies are required to explore the exact action mechanism of LiCl on 31 Table 1 for BHK cells and Table 2 The cytotoxicity of LiCl to BHK cells was detected by MTT assay. The result showed that within 169 the range of 25 mM LiCl, compared with the control group, the cell viability was ＞ 90% and 170 cell morphology have no significant change (Fig. 1) . Thus, 5-20 mM LiCl was selected in the 171 subsequent assays in vitro. 172 To investigate the replication of IBV in BHK cells, the CPE of BHK cells infected with IBV was 174 observed at different times. The main characteristic of BHK cells CPE included cells' shape 175 changed from spindle to round shape, or the cells gather or fall off to form visible plaque. The 176 significantly at 36h (Fig. 2A) . The viral load in BHK cells was measured by RT-qPCR. It was 178 found that when the BHK cells were infected at 36 h, the intracellular virus load got the highest 179 (Fig. 2B) . Next, the effect of LiCl on the replication of IBV in BHK cells was investigated. The 180 inhibitory effect of LiCl on IBV was determined by observing CPE and RT-qPCR. We found that 181 5 mM and 20 mM LiCl inhibited viral replication compared to mock-treated cells, especially when 182 the concentration of LiCl was 20 mM (Fig. 2C-D) . The results indicated that LiCl inhibited the 183 replication of IBV in a dose-dependent manner. 184 TNF-α, and IL-1β expression in BHK cells were determined using RT-qPCR. The data showed 196 that LiCl significantly blocked the expression of NF-κB, NLRP3, TNF-α, and IL-1β (Fig. 3B) . 197 These data demonstrated that LiCl inhibited inflammatory responses in IBV infected cells 198 Review of Infectious Bronchitis Virus Around the World S1 gene-based phylogeny of infectious bronchitis virus: An attempt to harmonize virus 270 classification The pathogenesis of nephritis in chickens induced by infectious bronchitis virus Transcriptome analysis of chicken kidney tissues following coronavirus 274 avian infectious bronchitis virus infection Nephropathogenicity of infectious bronchitis virus Coronavirus avian infectious bronchitis virus Infectious bronchitis. Diseases Of Poultry Vaccination against infectious bronchitis virus: A continuous challenge S1 gene sequence analysis of a 284 nephropathogenic strain of avian infectious bronchitis virus in Egypt Clinical use of lithium salts: guide for users and prescribers. 13. PC. B. 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