| objective:To investigate the effects of various doses of penehyclidine hydrochloride postconditioning on nuclear factorκB, and contents of TNF-α, IL-6 in rats with ALI induced by LPS, to explore the its therapy effects and possible mechanisms on rats with ALI.Methods:ALI models were established by intravenous injection of lipopolysaccharide. Sixty SD rats, weighed 220-280 g, were randomly divided and averaged into five groups, each group was twelve:control group (group NS), modle group (group LPS), LPS plus high dose of PHC group (group PH), LPS plus middle dose of PHC group (group PM), LPS plus low dose of PHC group (group PL). All rats received administration through caudal vein. NS group (n=12) was initially injected with saline (1ml), and received saline (1ml) injection 30 minutes later; LPS group (n=12) was initially injected with LPS (5mg/kg, saline diluted to 1ml), and received saline (1ml) injection 30 minutes later; PL, PM and PH group (n=12) were initially injected with LPS (5mg/kg, saline diluted to 1ml), and received PHC (0.3,1.0 and 3.0mg/kg, saline diluted to 1ml) injection respectively 30 minutes later. After 6 hours, they were anesthetized by 1% sodium pentobarbital (40 mg/kg) injected intraperitoneally, then the lung specimens were retrieved. NF-κB in Lung tissue were determined by the immunohistochemistry (IHC), The tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) were measured by ELISA, the lung water content, lung wet/dry weight (W/D) ratio was measured.Results:(1) Expression of NF-κB in the lung tissue in ALI rats. In control group, expression of NF-κB in lung tissue was weak. Expression of NF-κB in LPS group was significantly higher than that in control group (P<0.01). In PL, PM and PH group, expression of NF-κB was significantly lower than that in LPS group P<0.01). Expression of NF-κB in PL group was higher than that in PM and PH groups (P<0.01). Expression of NF-κB in PM group was lower than that in PH group (P<0.05). Expression of NF-κB in PH, PM and PL group was all higher than that in control group (P<0.05).(2) TNF-α and IL-6 contents in brochoalveolar lavage fluid in ALI rats. In control group, TNF-α and IL-6 contents in brochoalveolar lavage fluid were small in amount. TNF-α and IL-6 contents in LPS group were significantly higher than those in control group (P<0.01). In PL, PM and PH group, TNF-α and IL-6 contents were significantly lower than those in LPS group, TNF-α (P<0.05); IL-6 (P<0.01). TNF-α and IL-6 contents in PL group were higher than those in PM and PH group, TNF-α contents in PM group (P<0.01), TNF-α contents in PH group (P<0.05), IL-6 contents in PM and PH group (P<0.01). TNF-a and IL-6 contents in PM group were lower than those in PH group, TNF-α (P<0.05); IL-6 (P<0.01). TNF-α and IL-6 contents in PH, PM and PL group were all higher than those in control group (P<0.01).(3) Correlation analysis between expression of NF-κB protein and contents of TNF-α and IL-6. The expression of NF-κB protein was positively correlated with contents of TNF-α and IL-6 in ALI rats. r=0.70 and r=0.74 (P<0.01).(4) Correlation analysis between contents of TNF-a and IL-6. The contents of TNF-α were positively correlated with contents of IL-6 in brochoalveolar lavage fluid in ALI rats. r=0.86 (P<0.01).(5) The lung water content, wet/dry weight (W/D) ratio in lung tissue in ALI rats. In control group, the lung tissues were flexible, no edema, no abnormality in color. The lung water content, W/D ratio in LPS group was significantly higher than that in control group (P<0.01). In PL, PM and PH group, the lung water content, W/D ratio was significantly lower than that in LPS group (P<0.01). The lung water content, W/D ratio in PL group was higher than that in PM and PH group, the lung water content in PM group (P<0.05, the lung water content in PH group (P<0.01, W/D ratio in PM and PH group (P<0.01). The lung water content, W/D ratio in PM group was lower than that in PH group, the lung water content (P<0.01), W/D ratio (P<0.05). The lung water content, W/D ratio in PH, PM and PLgroup was all higher than that in control group (P<0.05).(6).The pathomorphological change of the lung tissues in ALI rats. HE showed that there were no pathological changes in the lung tissues of rats in control group. The lung tissues were infiltrated with plenty of inflammatory cells in LPS group. The alveolar spaces and interstitials were damaged severely and filled with erythrocytes and neutrophils in LPS group. In PL, PM and PH group, the lung injuries were less severe than those in the LPS group.Conclusions(1) Penehyclidine hydrochloride postconditioning could have protective effect on rats with ALI induced by LPS, this is partly due to its ability of inhibiting expression of NF-κB, lowering the levels of TNF-α and IL-6, eventually blocking the development of ALI by reducing inflammatory response.(2) The protective effect of 1.0 mg/kg penehyclidine hydrochloride postconditioning was best on rats with acute lung injury induced by endotoxin,3.0 mg/kg was second,0.3 mg/kg was weakest. |
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