| Objective To investigate the protective effect and mechanism of Xuebijing on acute lung injury(ALI)in rabbits induced by blast.Methods A total of 32 healthy and clean rabbits were divided into 4 groups by simple randomization: normal control group(NS),ALI group,low-dose Xuebijing group(L-XBJ)and high-dose Xuebijing group(H-XBJ),and each group has 8 rabbits.Injured rabbits by industrial explosive explosion by designated professional,after the explosion,some emergency treatments were implemented,and the general condition,mental state and stress reaction of the rabbits were observed and recorded.After the establishment of ALI animal model of rabbits induced by blast,the rabbits in L-XBJ group and H-XBJ group were intravenously injected with different doses of Xuebijing(10,20ml/kg),while the rabbits in NS group and ALI group were injected with normal saline.The blood was saved after 4 hours,and the bronchial alveolar lavage fluid(BALF)and lung tissue were collected at 24 hours;the MMP-9 and NF-k B in the lung tissue were evaluated by Western blot and the Reverse transcription polymerase chain reaction;the levels of Tumor necrosis factor-?(TNF-?,),Interleukin(IL-6,IL-10)in the BALF and serum were measured by ELISA methods;the wet to dry weight ratios(W/D)were determined;the pathological changes of lung tissues were observed under the light microscope.Result 1.General performance: Before the explosion,rabbits in each group had normal breathing,eating,drinking and activity.After the explosion,rabbits in ALI group,L-XBJ group and H-XBJ group had the following performance: breathing rate and heart rate are accelerated;dysphoria;the lip,ear and nose skin mucosa showed cyanosis;the oral and nasal mucosa appeared with bubble secretions;dry or wet rale appeared in Bilateral lung;with respiratory distress,a few rabbits will be unconscious.2.Performance of lung tissue under naked eyes: In NS group,the surface of the lung tissue was pink,smooth and elastic,intact,no damage and no liquid overflow.In ALI group,the surface of the lung tissue was dark red,the volume was significantly increased,the envelope was congested,and the surface of the lung showed multiple patchy hemorrhage foci.L-XBJ group and H-XBJ group lung tissues surface are also dark red,the degree of congestion and volume were lighter than that of ALI group.3.Determination of MMP-9 protein in lung tissue by Western blot: The NS group only a small amount of expression,protein expression was significantly higher in ALI group,L-XBJ group and H-XBJ group protein expression gradually decreased,with the H-XBJ group decreased significantly.4.Determination of MMP-9 m RNA expression in lung tissue by RT-PCR: The variance analysis showed that the expression level of MMP-9 m RNA was statistically significant(P<0.01).Compared with the NS group,ALI group expression was significantly higher(P<0.01),with the increase of the dose of Xuebijing injection,the expression level of L-XBJ group and H-XBJ group gradually decreased,with the H-XBJ group decreased significantly(P<0.01).5.Determination of NF-k B protein in lung tissue by Western blot: The NS group only a small amount of expression,protein expression was significantly higher in ALI group,L-XBJ group and H-XBJ group protein expression gradually decreased,with the H-XBJ group decreased significantly.6.Determination of NF-k B m RNA expression in lung tissue by RT-PCR: The variance analysis showed that the expression level of MMP-9 m RNA was statistically significant(P<0.01).Compared with the NS group,ALI group expression was significantly higher(P<0.01),with the increase of the dose of Xuebijing injection,the expression level of L-XBJ group and H-XBJ group gradually decreased,with the H-XBJ group decreased significantly(P<0.01).7.The wet to dry weight ratio(W/D)of lung tissue: The levels of W/D in ALI group,L-XBJ group and H-XBJ group were higher than the NS group,L-XBJ group and H-XBJ group decreased compared with ALI group,with the H-XBJ group decreased significantly(P<0.05).8.The levels of TNF-?,IL-6 in blood of the fourth hour: Compared with the NS group,The levels of TNF-?,IL-6 were significantly higher in ALI group(P<0.05),after Xuebijing treatment,L-XBJ group and H-XBJ group were lower than that of ALI group,H-XBJ group was reduced significantly(P<0.05).9.The levels of IL-10 in blood of the fourth hour: After Xuebijing treatment,L-XBJ group and H-XBJ group were higher than those in ALI group,and H-XBJ group were significantly higher(P<0.05).10.The levels of TNF-?,IL-6 in BALF: Compared with the NS group,The levels of TNF-?,IL-6 were significantly higher in ALI group(P<0.05),with the increase of the dose of Xuebijing,L-XBJ group and H-XBJ group were lower than that of ALI group,H-XBJ group was reduced significantly(P<0.05).11.The levels of IL-10 in BALF: With the increase of the dose of Xuebijing,L-XBJ group and H-XBJ group were higher than those in ALI group,and H-XBJ group were significantly higher(P<0.05).12.The pathological changes of lung tissue: The NS group was normal lung tissue,the structure was clear,the alveolar wall was complete,the alveolar cavity was clean,no inflammatory cell infiltration and interstitial edema fluid.Fractured alveolar wall,pulmonary alveolar and interstitial edema,a large of serous exudation,inflammatory cells,red blood cell leakage,occasionally transparent film,thickening of the alveolar septum fracture,different size of pulmonary alveoli,pulmonary atelectasis and emphysema can be seen in ALI group.In L-XBJ group and H-XBJ group,alveolar and pulmonary interstitial edema and inflammatory cell infiltration was significantly reduced compared with ALI group,H-XBJ group is most obvious.Conclusion 1.After the blast injury,the general performance of rabbits and the wet to dry weight ratio were combined with pathological changes,which suggested that the ALI model was successfully produced by explosion.2.The mechanism of Xuebijing which can inhibit lung inflammatory reaction and reduce the lung exudation of protein may be connected with blocking NF-?B inflammatory pathway,decrease the expression of MMP-9,and downregulating proinflammatory factor TNF-? and IL-6,upregulation of anti-inflammatory factor IL-10 levels,thereby reducing lung injury and protecting the lung function. |
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