Effects of penehyclidine hydrochloride on synthesis and release of HMGB1 in macrophages induced by lipopolysaccharide in mice

Objective To evaluate the effects of penehyclidine hydrochloride on synthesis and release of high mobility group protein box 1 (HMGB1) in macrophages induced by lipopolysaccharide (LPS) in mice.Methods RAW264.7 cells obtained from mice were seeded in the culture dishes.After being cultured for 24 h,the cells were randomly divided into 5 groups (n =6 each) using a random number table.The cells were incubated routinely in group C.The cells were incubated in the presence of LPS 500 ng/ml (group LPS),LPS 500 ng/ml + penehyclidine hydrochloride 0.5 μg/ml (group P1),LPS 500 ng/ml + penehyclidine hydrochloride 2 μg/ml (group P2),or LPS 500 ng/ml + penehyclidine hydrochloride 5 μg/ml (group P3).All the cells were incubated for 24 h.The cells and supernatant were collected.The proliferation of the cells was measured by CCK-8 assay,HMGB1 mRNA expression in the cells was detected by RT-PCR,NF-κBp65 protein expression in the cells was detected by Western blot and the concentration of HMGB1 in the supernatant was detected by ELISA.Results Compared with group C,the expression of HMGB1 mRNA and NF-κBp65 protein in the cells was significantly upregulated,and the concentration of HMGB1 in the supernatant was increased in LPS,P1,P2 and P3 groups (P ＜ 0.05).Compared with group LPS,the expression of HMGB1 mRNA and NF-κBp65 protein in the cells was significantly down-regulated,and the concentration of HMGB1 in the supernatant was decreased in P2 and P3 groups (P ＜ 0.05).Compared with group P1,the expression of HMGB1 mRNA and NF-κBp65 protein in the cells was significantly down-regulated,and the concentration of HMGB1 in the supernatant was decreased in P2 and P3 groups (P ＜ 0.05).There was no significant difference in the indicators mentioned above between group P2 and group P3 (P ＞ 0.05).There was no significant difference in the proliferation of the cells among the 5 groups (P ＞ 0.05).Conclusion Penehyclidine hydrochloride can reduce the synthesis and release of HMGB1 in macrophages induced by LPS through inhibiting NF-κB activation in mice. Key words: Cholinergic antagonists;  Lipopolysaccharides;  High mobility group proteins; Macrophages