1.Expression levels of interferon-α receptor, interferon-stimulated gene factor 3, double-stranded RNA-activated protein kinase and ribonuclease L in chronic hepatitis B patients treated with interferon
Qian YANG ; Xiaopeng LI ; Yuanbin ZHONG ; Tianxin XIANG ; Lunli ZHANG
Chinese Journal of Infectious Diseases 2020;38(11):711-716
Objective:To investigate the expression levels of interferon-α receptor (IFNAR), interferon-stimulated gene factor 3(ISGF3), double-stranded RNA-activated protein kinase(PKR) and ribonuclease L (RNase L) in patients with chronic hepatitis B (CHB) treated with interferon.Methods:From July 2014 to June 2017, 41 treatment naive CHB patients were enrolled in the Department of Infectious Diseases, First Affiliated Hospital of Nanchang University. Eighteen patients were treated with polyethylene glycol interferon α-2b, and 23 patients were treated with conventional interferon. The mRNA and protein expression levels of IFNAR1, IFNAR2, ISGF3, PKR and RNase L in peripheral blood mononuclear cells (PBMC) were detected by reverse transcription polymerase chain reaction and Western blot, respectively. The differences of these molecular expression levels in PBMC between the effective and ineffective groups were compared. The data were analyzed by t test. Results:After 24 weeks of treatment, 25 cases were effective, while 16 cases were ineffective. At four weeks of treatment, the mRNA expression levels of IFNAR1, IFNAR2 and PKR in PBMC of the effective group were 0.748±0.129, 1.169±0.125 and 1.047±0.091, respectively, which were all higher than those in the ineffective group (0.591±0.021, 0.689±0.059 and 0.791±0.033, respectively). The differences were statistically significant ( t=-4.304, 16.482 and -5.346, respectively, all P<0.01). The mRNA expressions of ISGF3 and RNase L in PBMC of the effective group were 0.739±0.159 and 0.780±0.140, respectively, while those in the ineffective group were 0.690±0.035 and 0.733±0.122, respectively, which were not significantly different ( t=-0.160 and -1.443, respectively, both P>0.05). The mRNA expression levels of IFNAR1, IFNAR2, ISGF3, PKR and RNase L at baseline, week eight, 12 and 24 of treatment in the effective group were all higher than those in the ineffective group (all P<0.01). The protein expression levels of IFNAR1, IFNAR2, ISGF3, PKR and RNase L in the effective group were all higher than those in the ineffective group (all P<0.01). Conclusion:After interferon treatment, the mRNA and protein expression levels of IFNAR1, IFNAR2, ISGF3, PKR and RNase L in PBMC of CHB patients are all increased, especially IFNAR2 and PKR levels increase in the early stage of treatment (four weeks).
2.Effects and mechanism of methionine restriction on macrophage for lipopolysaccharide-induced acute lung injury in mice
Xianjian LIAO ; Jing WEN ; Jiaxiang DUAN ; Lunli XIANG ; Zhen YANG ; Qingying HE ; Jiaolin NING
Journal of Army Medical University 2024;46(7):688-694
Objective To investigate the effects of methionine restriction(MR)on macrophages in lipopolysaccharide(LPS)-induced acute lung injury(ALI)and to explore the underlying mechanism.Methods According to the random number table method,36 male C57BL/6J mice(6~8 weeks old,23±2 g)were divided into 3 groups with 12 mice in each group:the sham group,the LPS group and the LPS+MR group.HE staining and pathological scoring of lung injury were performed in lung tissues.The expression of LPS-binding protein(LBP)and Toll-like receptor-4(TLR4)was detected by RT-qPCR and Western blotting.Macrophage-colony stimulating factor(M-CSF),granulocyte-macrophage-colony stimulating factor(GM-CSF)and chemokine C-C motif ligand 3(CCL3)which are all macrophage-associated chemokines were analyzed by immunohistochemistry.Results Compared with the sham group,the pathological score of lung injury in the LPS group was significantly increased(P<0.01);The mRNA and protein expression levels of LBP and TLR4 were significantly increased;The number of positive cells of CD11b,F4/80,M-CSF,GM-CSF and CCL3 were significantly increased(P<0.01).MR significantly improved LPS-induced ALI,and decreased the pathological score of lung injury(P<0.01);The mRNA and protein expression levels of LBP and TLR4 were decreased;Compared with the LPS group,the number of positive cells of CD11 b,F4/80,M-CSF,GM-CSF and CCL3 were reduced in the LPS+MR group(P<0.01).Conclusion MR could attenuate LPS-induced ALI by inhibiting the expression of macrophage chemokines and preventing infiltration and activation of macrophage to lungs.