The Protective Effect And Involved Mechanism Of Effective Fractions Of Xie-Xin Decoction On Severe Acute Panceatitis Associated Intestinal And Lung Injury

ObjectiveSevere acute pancreatitis (SAP) is a frequent acute abdomen in clinic, intestinal and lung injury is frequent complications, which still lack of effective therapy. Treatment effectiveness of integrated TCM-WM is better than conventional therapy of west medicine.Now, TCM is become one of important complementary therapies of SAP. Compatibility of free anthraquinone of rhubarb and total flavonoids of Scutellaria baicalensis Georgi is anti-endotoxic effective components from Xiexin Decoction. In this study, we arm to evaluate the potent effect of this compatibility on the the lung injury and intestinal injury of SAP, and more attention was payed to the underlying molecular mechanisms of this compatibility.MethodsThe SAP model was induced by standard retrograde infusion of bilio-pancreatic duct with 3.5% sterile sodium taurocholate solution at lml/kg in Sprague-Dawley rats, the same dose of saline solution, replacement of sodium taurocholate,were administrated in sham operation control group. Free anthraquinone of rhubarb, total flavonoios of Scutellaria baicalensis Georgi and their compatibility were administrated affusedly down alimentary canal from mouth to stomach at 200mg/kg within 8 and 2 hours before the inducement of SAP, rats were killed on scheme over a time course(3,6,12hours).The blood serum, small intestine and lung tissue of rats were harvested, H.E Dyeing of small intestine and lung tissue were employed for pathological assessment;the severity of tissue edema was determined by the ratio of the weight of wet tissue to that of dry tissue(ww/wd);the level of MPO,MDA and NO of small intestine and lung tissue homogenate were determined by method of zymochemistry;the content of EBD in small intestine and lung tissue,reflecting the microvascular permeability of tissue,was determined by Colorimetry, according to a standard curve of EBD;the level of serum TNF-a and IL-1βwere determined by radioimmunoassay(RIA);PTAH Dyeing of small intestine and lung tissue for microthrombus;the expression of COX-2, NF-κB and intercellular adherension molecule-1 (ICAM-1)protein were determined immunohistochemistrically,and expression of COX-2mRNA,iNOS mRNA,and TFmRNA in lung tissues were determined by RT-PCR.ResultsPART IThe SAP group, pathological assessment,ww/wd, level of MPO, MDA and NO,content of EBD were all significant increase, comparing with sham operation group(P<0.01).Free anthraquinone of rhubarb, total flavonoids of Scutellaria baicalensis Georgi and their compatibility had very excellent protective effect on lung and intestinal injury, they ameliorated the degree of pathological injuries about lung and intestine, decreased pathological score;they decreased the level of MPO, MDA and NO of small intestine and lung tissue homogenate significantly; the content of EBD and ww/wd were also decreased.The compatibility group had more excellent protective effect than other groups.PART IIThe SAP group, serum TNF-a and IL-1βwere significant increase; microthrombus determined by PTAH could be observed in tissue;significant overexpression of NF-κB, COX-2 and ICAM-1 were demonstrated in lung and intestinal tissue of rat with SAP by IHC; COX-2mRNA, iNOS mRNA and TFmRNA were also overexpression in lung and intestinal tissue of rat with SAP by RT-PCR. The compatibility obviously decreased the level of serum TNF-a and IL-1β,decreased the density of microthrombus;it inhibited the expression of NF-κB, ICAM-1,and COX-2 protein in lung and intestinal tissue over the time course, except the expression of COX-2 in intestinal tissue at 12hour and the expression of ICAM-1 in lung tissue at 3 and 12hour, in intestinal tissue at 3 and 6hour;it dramatically down-regulated the expression of COX-2mRNA,iNOS mRNA and TFmRNA in lung and intestinal tissue.Conclusions(1)Our results showed that obvious intestinal and lung injury were observed after SAP model was induced 3h, it had significant correlation between intestinal injury and lung injury. The studies suggested that additional attention should be paided to treatment of intestinal and lung injury,except for conventional therapy.(2)Our results clearly suggested that free anthraquinone of rhubarb,total flavonoids of Scutellaria baicalensis Georgi and their compatibility reduced the degree of serum AMS, ameliorated the degree of pathological injury of intestinal and lung,reduced the content of MPO, NO, MDA and EBD of intestinal and lung tissue.The compatibility group had more excellent protective effect on lung and intestinal injury than other groups, the result of compatibility was increase efficiency(3)The compatibility reduced degree.of injury of.inflammationary and oxidative stress by decreasing stagnation of polymorphonuclear leukocytes in lung and intestinaltissue of SAP rats,but the effect might not by inhibiting the expression of ICAM-1 to attain.The compatibility ameliorated microvascular permeability and microcirculation disturbance of lung and intestinal tissue of SAP rats, the effect was close relevant to suppression of COX-2 and TF activation, which might be important mechanism about ameliorating microcirculation disturbance of tissue.The compatibility alleviated drgree of systemic inflammatory reaction and organ function impairment by inhibiting TNF-α,IL-1βand NO. The compatibility significantly down-regulated the expression of NF-κB,which was a main nuclear factor about regulating the transcription of TNF-α,COX-2,iNOS and ICAM-1,and so on.Thereby, we presumed that the compatibility depressed the expression of factors above mentioned via suppression of NF-κB activation.