title: [Inhibitory effect of kukoamine B on lung inflammatory responses in mice with sepsis].
abstract: OBJECTIVE
  To investigate the inhibitory effect of kukoamine B (KB) on lung inflammatory responses in mice with sepsis and its possible molecular mechanism.
  
  
  METHODS
  Twenty-eight male mice were randomly divided into control group (n=8), lipopolysaccharide (LPS) group (n=10), and LPS + KB group (n=10). Sepsis model was reproduced by intra-peritoneal injection of 20 mg/kg LPS, while equivalent normal saline was given in control group, and 20 μg/kg KB was injected through caudal vein 4 hours after LPS challenge in LPS + KB group. After 8 hours of LPS challenge, the concentration of LPS in plasma and the activity of myeloperoxidase (MPO) in the lung tissue were determined. The contents of tumor necrosis factor-α (TNF-α) and interleukin-1β(IL-1β) in plasma, alveolar lavage fluid and lung tissue homogenates were assessed by enzyme linked immunosorbent assay (ELISA). The activation of nuclear factor-ΚB (NF-ΚB) and the expression of inducible nitric oxide synthase (iNOS) in lung tissue were determined by Western Blot. The pathological changes in lung tissues were observed with hematoxylin-eosin (HE) staining. The expression of intercellular adhesion molecule-1 (ICAM-1) in lung tissue was determined by immunohistochemistry.
  
  
  RESULTS
  Compared with control group, the concentration of LPS in plasma (1 155.650±147.149 kEU/L vs. 31.390±18.859 kEU/L), MPO activity (1.177±0.093 U/g vs. 0.775±0.166 U/g), NF-ΚB activity (gray value: 1.557±0.105 vs. 0.824±0.032) and the expression of iNOS (gray value: 0.650±0.129 vs. 0.392±0.097) were significantly increased in LPS group (all P<0.05). After KB intervention, the concentration of LPS (624.461±149.012 kEU/L), MPO activity (0.919±0.023 U/g), NF-ΚB activity (1.127±0.074) and the expression of iNOS (0.425±0.066) were significantly lowered (all P<0.05). Compared with control group, the contents of TNF-α (47.325±13.864 ng/L vs. 6.534±0.544 ng/L, 13.382±2.231 ng/L vs. 3.748±0.692 ng/L, 31.127±7.399 ng/L vs. 14.948±4.673 ng/L) and IL-1β (74.329±11.890 ng/L vs. 29.921±6.487 ng/L, 9.422±2.674 ng/L vs. 1.105±0.364 ng/L, 528.509±32.073 ng/L vs. 109.945±13.561 ng/L) in plasma, alveolar lavage fluid and lung tissue homogenates were obviously enhanced in LPS group (all P<0.05). With KB intervention, the contents of TNF-α (20.331±7.789 ng/L, 7.145±1.202 ng/L, 15.966±2.946 ng/L) and IL-1β (57.707±8.098 ng/L, 2.212±0.878 ng/L, 426.154±11.270 ng/L) were markedly reduced (plasma TNF-α: F=16.052, P=0.002; IL-1β: F=20.649, P=0.000; lung tissue homogenates TNF-α: F=31.134, P=0.001; IL-1β: F=22.792, P=0.002; alveolar lavage fluid TNF-α: F=10.013, P=0.009; IL-1β: F=319.857, P=0.000). In addition, leukocyte infiltration to the lung tissue was attenuated, and the expression of ICAM-1 was reduced by KB in histological examination.
  
  
  CONCLUSIONS
  KB, as a neutralizer of LPS, can inhibit the release of inflammatory mediators, reduce the pulmonary inflammatory response and protect the function of lung in septic mice.
authors.affiliation: Department of Burns and Plastic Surgery, Affiliated Hospital, Jiangsu University, Zhenjiang 212001, Jiangsu, China. Corresponding author: Sun Bingwei, Email: sunbinwe@hotmail.com.
authors.forename: Jinli
authors.initials: J
authors.lastname: Zhang
authors.forename: Weiting
authors.initials: W
authors.lastname: Qin
authors.forename: Wanghui
authors.initials: W
authors.lastname: Lyu
authors.forename: Weichang
authors.initials: W
authors.lastname: Shen
authors.forename: Xu
authors.initials: X
authors.lastname: Wang
authors.forename: Bingwei
authors.initials: B
authors.lastname: Sun
mesh_terms: D000818:Animals; D002109:Caffeic Acids / Q000494:pharmacology*; D004195:Disease Models, Animal; D053583:Interleukin-1beta / Q000378:metabolism; D008168:Lung / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D016328:NF-kappa B / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D009195:Peroxidase / Q000378:metabolism; D018805:Sepsis / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D013096:Spermine / Q000031:analogs & derivatives* / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D002109:Caffeic Acids; D053583:Interleukin-1beta; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; C557784:kukoamine B; D013096:Spermine; D009195:Peroxidase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse