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. 2022 Jan:162:105352.
doi: 10.1016/j.micpath.2021.105352. Epub 2021 Dec 7.

Lithium chloride inhibits infectious bronchitis virus-induced apoptosis and inflammation

Xingyun Liu  1 , Xinyu Chang  1 , Qin Wu  1 , Jun Xu  1 , Lu Chen  1 , Ruiting Shen  1 , Xiaolin Hou  2
Affiliations

Lithium chloride inhibits infectious bronchitis virus-induced apoptosis and inflammation

Xingyun Liu et al. Microb Pathog. 2022 Jan.

Abstract

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.

Keywords: Apoptosis; Chicken embryos; IBV; Inflammation; LiCl; Survival rate.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1
Fig. 1
Cytotoxic effect of LiCl on BHK cells. MTT assay was used to detect the cytotoxicity of LiCl to BHK cells. The survival rates of BHK cells were given at different concentrations of LiCl, and cell survival rates of more than 90% were considered to be the maximum non-toxic concentration of LiCl. Data are shown as the mean ± SEM of three independent experiments.
Fig. 2
Fig. 2
LiCl inhibited cytopathy and IBV replication. The BHK cells were with 10 TCID50 IBV. (A) The CPE of BHK cells infected with IBV at 24 h.p.i and 36 h.p.i. (B) The intracellular viral copies at specified times after BHK cells infected with IBV. (C) Therapeutic effects of different concentrations of LiCl on IBV-induced CPE. (D) The viral copies in cell lysates with different concentrations of LiCl. Viral RNA was extracted and quantified with RT-qPCR. The data are expressed as mean ± SEM of 3 independent experiments. ###p < 0.001, compared with the mock group. ***p < 0.001, compared with the IBV infection group.
Fig. 3
Fig. 3
LiCl inhibited IBV-induced apoptosis and inflammation in vitro. The BHK cells were mock-treated or treated with different concentrations LiCl for 36 h after infection with 10 TCID50 for 1h. Total RNA was subsequently extracted from cell lysates. (A) The mRNA levels of Caspase-3, Bax and Bcl-2 were assessed by RT-qPCR. (B) The mRNA levels of NF-κB, NLRP3, TNF-α and IL-β were assessed by RT-qPCR. The data were expressed as mean ± SEM of 3 independent experiments. ###p < 0.001, compared with the mock group. ***p < 0.001, compared with the IBV infection group.
Fig. 4
Fig. 4
LiCl inhibited IBV-induced apoptosis and inflammation in chicken embryos Chicken embryos infected with 100 TCID50 IBV were injected with different concentrations of LiCl for treatment. Allantoic fluid and CAM were collected at 72 h.p.i after infection. (A) The levels of IBV-N in the allantoic fluid were analyzed by RT-qPCR. (B–C) The levels of apoptosis and inflammation-related genes in CAM were measured by RT-qPCR. (D) The survival rate of chicken embryos was recorded every day until the 9th day. (E) The embryo's growth of each group on the 9th day. The data were expressed as mean ± SEM of 3 independent experiments. ###p < 0.001, compared with the mock group. ***p < 0.001, compared with the IBV infection group.
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