Toxins Clear Klebsiella Pneumoniae Pneumonia Caused By The Mods Aged Rats The Jak / Stat Signal Transduction Of The Intervention Effect

(Multiple organ dysfunction syndrome,MODS) MODS often happens to the aged men in clinical crisis and is an important reason for their death.Mediators of inflammation excessive expression is the main function mechanism of MODS and related to multiple signal passage activation. TNF-a,IL-1 and IL-6 are the key factors to trigger and induce excessive inflammatory response by cascade amplification and the main medium for SIRS , whose composition and release is closely related to JAK/STAT signal pathway. JAK/STAT signal pathway play a very important role in the MODS and blockage it to lessen the organ injury. Chinese medicine call multiple organ failure as Zang-organ failure syndrome.Its pathogenesis is pyrotoxin invasion, haemostasis blockage, qi yin insufficiency, three of which interact to cause negative results. Therefore, clearing away heat and toxic material, promoting blood circulation by removing blood stasis, supplementing qi and nourishing yin are main treatments for multiple organ failure, by which Du Su Qing granule has been developed. Animal experiment and clinical observation suggest the positive effect of Du Su Qing to treat senile pneumonia and Du Su Qing can protect other organs against injury, imrove the patient's living condition and lower death rate significantly. This experiment duplicates MODS model in rats to explore the pathological and physiological characteristics of rats with MODS and the effects of ageing factor on aged rats in biochemistry and pathology and observe the effects of Du Su Qing on aged rats in biochemistry , pathology and physiology and JAK/STAT passage and their defense mechanism.1 Objective To observe the pathological and physiological characteristics of rats with MODS and the effects of ageing factor on lung, heart, kidney and intestine; To observe the effects of the multiple organ dysfunction in rats on inflammatory cytokine , JAK/STAT signal pathway and explore the relationship between IL-1,IL-6,TNF-a and JAK/STAT signal pathway. To observe of Du Su Qing on multiple organ dysfunction in aged rats on pathology and biochemistry; to probe its defense mechanism to lung, heart, kidney and intestine and possible mechanism ; to observe the effect of Du Su Qing on multiple organ dysfunction in aged rats in JAK/STATsignal passage; to probe its defense protection against multiple organ injury and possible mechamism.2 Method We put the SD aged rats into 8 groups and the SD young rats into 2 groups. Building a multiple dysfunction model by tracheal intubation in klebsiella pneumoniae pneumonia aged rats and kill them 48hours later. We take the lung tissues, do bronch-oalveolar lavage, identify the bacteria type, test the water content in lung, heart, liver, kidney and intestine. Sampleing the rat peripheral blood, observe the ratio between leukocyte and neutrophil, the changes of blood gas index and study the pathological and microstructure changes in rats by the pathomorphological method and measure the related chemical index and the effect of Du Su Qing granule on these changes. We observe the protein expression changes of IL-1, IL-6, TNF-a, STAT1, STAT3,STAT5,JAK2,MPO in lung, heart, kidney, intestine in aged and young rats caused by klebsiella imeumoniae multiple organ dysfunction and the effects of Du Su Qing granule on these expressions.3 Results3.1 Duplicated the model of MODS successfullyThrough the comparison between model group rats and control group rats, leuco- cytes is higher,total white blood cells increase, the ratio of neutrophlic granulocytes rises. PaCO2 in lung is higher , PaO2 is lower. LDH and CK are much higher in heart(P<0.01).Liver function suggests ALT and AST is higher(P<0.05).It is observed that the loss villiof the small intestinal heighten obviously,Inflammatory cell infiltration in subnucosa angiote-lectasis and capillary injection. Bi-visceral dysfunction accounts for 40% in rats, triple-visceral dysfunction is 60%. Pathological injury grade numerical score of lung, heart, liver, intestine in model group is much higher than that in control group. Animal model of the multiple organ dysfunction has been duplicated successfully to the standard of animal model of MODS.3.2 The characteristics of lung, heart, kidney, intestine of MODS aged rats in pathology and physiology and their effects on JAK/STAT signal pathway.The difference of rat lung injury between young control groups and aged control groups is pathologically much higher (P<0.01) and the injury degree of aged model groups is much heavier than that of young model groups (P<0.01). Compared to control groups, MPO, IL-1, IL-6, TNF-a, JAK2, STAT1, STAT3, STAT5 are expressed highly(P<0.01) ,JAK2mNRA expression is up-regulated (P<0.01), MPO, IL-1, STAT1, STAT5 positive cells or the average optical density of protein expression in aged model groups are higher or much higher than that in young model groups (P<0.05 or P<0.01).The pathological injury in model groups is obvious, and we can see cardiac muscle fibers disorder and a small or large amount of inflammatory cellular infiltration, LDH,CK,CK-MB is higher in the enzyme of the cardiac (p<0.01). IL-1, IL-6, TNF-a, JAK2, STAT1, STAT3, STAT5 in model groups are highly expressed (p<0.01), JAK2mNRA expression is up-regulated (p<0.01), compared to young model groups, TNF-a,STAT1,STAT5,JAK2 positive cells or the average optical density of protein expression in aged model groups are higher or much higher than that in young model groups (P<0.05 or P<0.01).The difference of rat liver injury between young control groups and aged control groups is pathologically higher (P<0.05) and pathological changes are main liver cell vacuolar-degeneration. ALTand AST are much higher in model groups(p<0.01).Model groups show certain injuries, especially in nephric tubele.There is no obvious difference in the changes of BUN and Cr between model groups and control groups. IL-1,IL-6,TNF-a,JAK2,STAT1,STAT3,STAT5 are highly expressed(P<0.01)in model groups. The protein expression of IL-6,STAT3,STAT5 in aged control group is higher or much higher than that in young control group (P<0.05 or P<0.01),IL-1,STAT1,STAT3 positive cells or the average optical density of protein expression in aged model groups are higher or much higher than that in young model groups (P<0.05 or P<0.01).3.3 The effects of Du Su Qing on lung, heart, liver, intestine of MOD aged rats in pathology and physiology and its effects on JAK/STAT signal pathway.The difference of rat lung injuries in aged control groups is pathologically much higher (P<0.01), total white blood cells increase,lung water content is much higher, PaCO2 is higher and PaO2 is much lower (P<0.01) in serum. IL-1,IL-6,TNF-a,JAK2,STAT1,STAT3,STAT5 are highly expressed(P<0.01)in aged model groups. AK2mNRA expression is up-regulated (p<0.01), all dose groups show PaCO2 is lower, PaO2 higher(p<0.01), injury degrees are improved(p<0.01), The expression of MPO,IL-1,IL-6 and TNF-a is much lower (P<0.01) ,expression of JAK2m(except for Lomefloxacin) STAT1,STAT3,STAT5 is down–regulated. Du Su Qing group, AG490group , AG490unit group ,Rapamycin group can down-regulate JAK2mNRA expression ( P < 0.01 ) , up-regulate SOCS3mNRA expression.The rat death rate in Du Su Qing group is lower than that in Lomefloxacin group, injury degree score is lowered and Du Su Qing group is optimal to Lomefloxacin group in lowering MPO,IL-1,TNF-a,STAT1,STAT3 and STAT5 expressin(P<0.01), but AG 490 union group and rapanycin group are optimal to Du Su Qing group, AG490 group and Rapamycin group in up-regulating MPO,IL-1,IL-6 and TNF-a protein expression.Rapamycin group can obviously lower STAT1,STAT3,STAT5 expression which is more significant(p<0.01) compared to Du Su Qing group, AG490 group and Rapamycin group. AG490 group and Rapamycin group are optimal to Du Su Qing group in lowering IL-1 expression.In aged model group rat myocardial injury has significant difference (p<0.01), water content is higher, LDH,CK and CK-MB in serum rise(P<0.01), rats IL-1, IL-6, TNF-a, JAK2, STAT1,STAT3 and STAT5 in aged model groups are high expressed (P<0.01), JAK2mNRA is up-regulated(P<0.01), each dose group can obviously lower LDH and CK-MB, improve injury degree(P<0.01), and reduce the expression of IL-1,IL-6(except for Lomefloxacin ) and TNF-a, down–regulate the protein expression of STAT1,STAT3,STAT5(except for Lomefloxacin). All dose group can down-regulate JAK2mNRA gen (P<0.01),AG490uniou group can up-regulate SOCS3mNRA gene expression. The rat death rate in Du Su Qing group is lower than that in Lomefloxacin group, injury degree score is lowered, but which have no significance in statistics; Compared to Lomefloxacin group, Du Su Qing grop can significantly lower the expression of STAT1,STAT3 and STAT5(P<0.01). AG490 union group and Rapamycin group is optimal to Du Su Qinggrop,AG490grop and Rapamycin grop in down-regulating the protein of IL-1,STAT1,STAT3 and STAT5. Besides, Rapamycin union group can significantly lower IL-6 expression, which is more significant(p<0.01) compared to Du Su Qing group,AG490 group and Rapamycin group. AG490 group and Rapamycin group are optimal to Du sSu Qing group in reducing the expression of IL-1,TNF-a,STAT1,STAT3 and STAT5. The difference of rat liver injuries in aged control groups is pathologically much higher (P<0.01), water content increases, ASTand ALT in serum rises,. All dose grops can obviously lower LDH,CK and CK-MB. Compared to model group,AG490 pathological injury score is significant (P<0.01); the pathological injury scores of remaining groups tend to decrease, but compared to model group there is no significance. All dose groups have no obvious differences in lowering LDH,CK and CK-MB.Rat kidney injuries in aged model groups pathologically worsen, death rate rises, water content increases, BUN and Cr (P<0.01) in serum decrease , but which is insignificant compared to control group. IL-1, IL-6, TNF-a, JAK2, STAT1, STAT3, STAT5 are highly expressed(P<0.01)in aged model groups. AK2mNRA is up-regulated (p<0.01);all dose groups can lower BUN and Cr, improve pathological injury degree which is not significant compared to model group. Compared to model grops, Lomefloxacin groups show no difference in decreasing the expression of IL-1,IL-6,TNF-a and JAK2, but the remaining group can significantly reduce the expression of IL-1,IL-6,TNF-a, down-regulate the protein expression of STAT1,STAT3,STAT5 and JAK2.The rat death rate in Du Su Qing group is lower than that in Lomefloxacin group, pathological injury scores decrease, which are not significant statistically; compared to Lomefloxacin group, Du Su Qing grop can significantly lower the expression of IL-1, IL-6, TNF-a, JAK2, STAT1, STAT3 and STAT5(P<0.01).AG490group , Rapamycin group and Du Su Qing group show no significant difference in lowering the protein expression of inflammatory cytokines.Rat intestine injuries in aged model groups worsen pathologically, death rate rises, water content increases.IL-1,IL-6,TNF-a,JAK2,STAT1,STAT3 and STAT5 in rats are highly expressed(P<0.01)in aged model groups. AK2mNRA and SOCS3mNRA are up-regulated (p<0.01); all dose groups can improve the intestine pathological injury degree, which has significant difference from model group(P<0.05或P<0.01). Model groups and Lomefloxacin groups show no difference in the decrease the expression of IL-6 and JAK2, but the rest groups can see the significant decrease of the expression of IL-1,IL-6 and TNF-a , the down-regulation of the protein expression of STAT1,STAT3,STAT5 and JAK2 and the down-regulation of JAK2mNRA(P<0.01 orP<0.05).The rat death rate in Du Su Qing group is lower than that in Lomefloxacin group and pathological injury scores decrease, which are not significant statistically; compared to Lomefloxacin group, Du Su Qing grop can significantly lower the expression of IL-1, IL-6, JAK2, STAT1, STAT3 and STAT5(P<0.01).AG490 union group is optimal to Rapamycin group and Du Su Qing group in down-regulating the protein expression of IL-1, IL-6, JAK2, STAT1, STAT3,STAT5 and SOCS3mNRA expression (P<0.01) and optimal to Rapamycin group in down-regulating IL-1 protein expression and SOCS3mNRA gene expression. Rapamycin group is optimal to Du Su Qing group and Lomefloxacin group in lowering the expression of IL-1, IL-6, TNF-a, STAT1, STAT3,STAT5,JAK2 and SOCS3mNRA (P<0.01) and optimal to Rapamycin group and AG490 group in down-regulating IL-6,STAT1and STAT3 protein expression. AG490 group and Rapamycin group are optimal to Du Su Qing group and Lomefloxacin group in lowering IL-6,STAT1,STAT5 protein expression. In addition, compared to Du Su Qing group, Rapamycin group can significantly lower STAT3 expression and AG490 group can significantly lower SOCS3mNRA expression.4 ConclusionMultiple organ dysfunction model in rats with klebsiella imeumoniae was successfully duplicated. Compared to young rat model, rat death rate is higher and pathological injury degree is more serious. Inflammatory cytokines played an important role in MODS pathological process. JAK/STAT signal passage mediated the amplification of imflammatory, and increased inflammatory injury to organs, caused MODS. Du Su Qing lowered the related cytokine expression, weakened JAK/STAT signal passage activation. Its defense mechanism to MODS probably blocked JAK/STAT signal passage, and had synergistic function with blocker to up-regulate the cytokine expression and protect the organs from injuries.