The Influence Of Intestinal Lymphatic Ligation And Traditional Chinese Medicine In The Yangming Fushi Induced Pulmonary Injury Rat

Abstract:Objective:(1) observe the influence of Intestinal lymphatic ligation in the Yangming Fushi induced pulmonary injury rat. (2) study the influence of traditional Chinese medicine in the Yangming Fushi induced pulmonary injury rat.(3)the involved mechanism of Intestinal lymphatic ligation and traditional Chinese medicine in the Yangming Fushi induced pulmonary injury rat.Methods:(1) male Sprage-Dawley(SD) rats were randomly divided into:the saline group, saline+intestinal lymph duct ligation group, Yangming Fushi model group and Yangming Fushi+ intestinal lymph duct ligation group. The sever actue pancreatitis was induced by retrograde infusion of bilio-pancreatitis duct with 3.5% sterile sodium tanrocholate solution.So we got the Yangming Fushi model.Isolated mesenteric lymph duct, silk ligated prepare saline+ intestinal lymph duct ligation group and Yangming Fushi+ intestinal lymph duct ligation group.Respectively,6h,12h,24h at different time points after the model is successful, take the portal vein and abdominal aortic blood, separate plasma and serum;draw pulmonary tissue, part of the preparation of tissue homogenates, partially embedded, sliced and HE staining.Azo chromogenic substrates within the Limulus test quantitative method to detect rat plasma endotoxin (ET) levels; Griess method of measuring serum nitric oxide(NO) levels; enzyme-linked immunosorbent assay(ELISA) measured the levels of tumor necrosis factor-a (TNF-α), interleukin -1 (IL-1) in serum pulmonary tissue homogenate;enzyme myeloperoxidase detect the level of myeloperoxidase (MPO) and enzyme superoxide dismutase detect the level of superoxide dismutase (SOD) in pulmonary tissue homogenate. Light microscopy of pulmonary histopathological changes. (2)male SD rats were randomly divided into:saline group, Yangming Fushi model group and Yangming Fushi+ free anthraquinone of rhubarb group; Free anthraquinone of rhubarb(20g/kg) was administrated affusedly down alimentary canal from mouth to stomach with 2ml at 2 hours before and after the inducement of Yangming Fushi model.Respectively,6h,12h,24h, take the rat arterial and venous blood, and pulmonary tissue, part of the preparation of tissue homogenates, partially embedded, sliced and HE staining. Methods and indicators with a method (1). (3)male Sprage-Dawley(SD) rats were randomly divided into:the saline group, Yangming Fushi model group,Yangming Fushi + intestinal lymph duct ligation group and Yangming Fushi+ free anthraquinone of rhubarb.Collect the pulmonary tissue of every rat after 24h. The pulmonary tissuepartially embedded and sliced.The expression of NF-κB and ICAM-1 protein were determined by Immunohistochemical method.The expression of iNOS mRNA and TLR4mRNAwere determined by RT-PCR.Results:(1)The portal vein:compared with Yangming Fushi model group, the level of plasma ET in Yangming Fushi+ intestinal lymph duct ligation group was decreased(P<0.05).the levels of serum NO, TNF-a and IL-1in 12、24h in Yangming Fushi +intestinal lymph duct ligation group was significantly decreased(P<0.01).the level of plasma ET in 12、24h and the level of serum NO in 6h in Yangming Fushi+ free anthraquinone of rhubarb group was decreased(P<0.05). the levels of serum TNF-α and IL-li in Yangming Fushi +free anthraquinone of rhubarb group was significantly decreased(P <0.01).(2)Arterial blood:compared with Yangming Fushi model group, the level of plasma ET in Yangming Fushi+ intestinal lymph duct ligation group was decreased(P<0.05).the level of serum IL-1 in 12、24h and the levels of serum TNF-α and NO were significantly decreased(P<0.01).the level of plasma ET and the level of serum IL-1 in 6h in Yangming Fushi+ free anthraquinone of rhubarb group was decreased(P<0.05).the level of serum IL-1 in 6、12h and the levels of serum TNF-α and NO were significantly decreased(P <0.01).(3)pulmonary tissue homogenates:compared with Yangming Fushi model group,the levels of serum TNF-α and IL-lin 12、24h and the level of serum MPO in 6、12h in Yangming Fushi+ intestinal lymph duct ligation group were significantly decreased(P<0.01).the level of serum MPO was increased(P<0.05).the levels of serum TNF-α、IL-1and MPO in Yangming Fushi+free anthraquinone of rhubarb group were significantly increased(P<0.01).the level of serum MPO in 12h was increased(P<0.05).Light microscopy histological changes in the lungs of rats, pulmonary injury in rats significantly improved in both Yangming Fushi+ intestinal lymph duct ligation group and Yangming Fushi+ free anthraquinone of rhubarb group.Furthermore the inflammation of them is reduced than Yangming Fushi group.(4)the study of mechanism:the expression of NF-κB and ICAM-1 protein were up-regulated in Yangming Fushi model by Immunohistochemical method; the expression of NF-κB and ICAM-1 protein were down-regulated in both Yangming Fushi+ intestinal lymph duct ligation group and Yangming Fushi+ free anthraquinone of rhubarb group.The expression of iNOS mRNA and TLR4mRNA were increased in Yangming Fushi model by RT-PCR.the expression of iNOS mRNA and TLR4mRNA were down-regulated in both Yangming Fushi+ intestinal lymph duct ligation group and Yangming Fushi+ free anthraquinone of rhubarb group.Conclusions:(1)The intestinal lymph duct ligation prevents endotoxin translocation and the release of immflammation factors,such as NO, TNF-a and IL-1.Reduce the level of PMN and the level of MPO was decreased.Reduce the immflammation injury.It can specifically scavening superoxide anion that protect the body from oxidative damage.(2)The free anthraquinone of rhubarb prevents endotoxin translocation and the release of immflammation factors,such as NO, TNF-α and IL-1.Reduce the level of PMN and the level of MPO was down-regulated.Reduce the immflammation injury.It can specifically scavening superoxide anion that protect the body from oxidative damage.the expression of NF-κB and ICAM-1 protein were down-regulated in both Yangming Fushi+ intestinal lymph duct ligation group and Yangming Fushi+ free anthraquinone of rhubarb group.The expression of iNOS mRNA and TLR4 mRNA were prevented too.the effect of depressing immflammation factors might not by inhibiting the expression of ICAM-1 to attain.NF-KB was a crucial nuclear factor about regulating the transcription of ICAM-1、iNOS and so on.Thereby,we presumed the mecanism of both intestinal lymph duct ligation and free anthraquinone of rhubarb inflence the rats with Yangming Fushi induced pulmonary injury via supression of NF-κB activation.