Protective effect of panaxadiols against brain injury induced by LPS through inhibiting NOS activity and p38 expression

AIM: To explore the protective effect of panaxadiols (PDS) on brain injury induced by endotoxin and its mechanism. METHODS: Rats were divided into control,LPS,LPS+dexamethasone (DEX) and LPS+PDS group, respectively. NOS activity, NO content and phosphorylated p38 expression in brain cortex were assayed 4 h after intravenous injection of LPS. RESULTS: NOS activity, NO content and phosphorylated p38 expression in brain cortex in LPS group were obviously higher than those in LPS group. NOS activity, NO content and phosphorylated p38 expression in brain cortex in LPS+DEX and LPS +PDS groups were obviously lower than those in LPS group. CONCLUSION: The protective effects of PDS against brain injury induced endotoxin may be related to decreasing NOS activity, NO content in the brain tissue, and this process is involved in p38MAPKs signal transduction.     

Effects of PDS on TLR4 mRNA expression in cortex in endotoxic shock rats

AIM: To explore the molecular mechanism of brain tissue injury induced by endotoxin. METHODS: Rats were divided into LPS, LPS+DEX, LPS+PDS and control group, respectively. NOS activity, NO content and TLR4 mRNA expression were assayed 4 h after intravenous injection of LPS. RESULTS: NOS activity, NO content and TLR4 mRNA expression in LPS+DEX and LPS +PDS groups were obviously lower than those in LPS group. CONCLUSION: PDS may provide protective effects on the central nervous system by down-regulating TLR4 expression, reducing NOS activity and NO content in the brain tissue.     

Paeoniflorin attenuates LPS-induced acute lung injury in mice

AIM: To investigate the mechanisms by which paeoniflorin (Pae) attenuates lipopolysaccharide(LPS)-induced acute lung injury in mice. METHODS: Male BALB/c mice were randomly divided into 4 groups: control,LPS, Pae+LPS, and Pae. Mice were administered intragastrically with double distilled water or Pae (20 mg/kg) once a day for 3 days. One hour after intragastrical treatment on the third day, LPS (20 mg/kg) or normal saline was injected intraperitoneally. Twelve hours after LPS challenge, the histological changes of the lung were observed, and histology score was also assessed. The myeloperoxidase (MPO),cytosolic phospholipase A₂ (cPLA₂) and phosphorylated cytosolic phospholipase A₂ (phospho-cPLA₂) in lung tissues were detected by Western blotting.RESULTS: LPS challenge resulted in acute lung injury, activated cPLA₂ and increased MPO content in lung. Pretreatment with paeoniflorin significantly attenuated lung injury induced by intraperitoneal injection of LPS. The levels of MPO and phospho-cPLA₂ in the lung tissues of the mice in Pae+LPS group were lower than those in LPS group (P<0.05).CONCLUSION: Pretreatment with paeoniflorin remarkably reduces LPS-induced acute lung injury through inhibiting phosphorylation of cPLA₂ and decreasing neutrophil infiltration in the lung. These findings provide a new strategy for the prevention and treatment of LPS-induced acute lung injury.     

Effect of panaxadiols on AQP1 expression in lungs of LPS shock rats

AIM：To investigate the alteration of aquaporin 1 (AQP1) expression in lung tissues in endotoxic shock rats and the effect of panaxadiols.METHODS：The histological changes were examined by HE staining.The expression of AQP1 was analyzed by immunohistochemical staining and Western blotting.RESULTS：(1) Significant inflammatory changes in pulmonary interstitial in rats of LPS group were observed.However,in dexamethasone (DEX) treatment group and panaxadiols (PDS) treatment group,the pulmonary pathologic changes were much slighter.(2) Positive AQP expression in the endothelium of capillary vessels on the wall of alveolus were seen in control (CTR) group,while merely very weak expression in LPS group was detected.The positive staining in PDS groups and DEX group were significantly stronger than that in LPS group,and close to CTR group.(3) The results of Western blotting showed that the quantity of AQP1 expression in LPS group was significantly lower than that in other groups,also the expression in PDS groups and DEX group was slightly lower than that in control group.CONCLUSION：LPS inhibits the expression of AQP1,while PDS increases the strength of AQP1 expression in lungs of endotoxic shock rats.     

Berberine prevents cardiac dysfunction and myocardial neutrophil accumulation independent of α2 adrenoceptor in endotoxemic mice

AIM: Berberine (Ber) has been reported to prevent lipopolysaccharide (LPS)-induced cardiac dysfunction, reduce neutrophil infiltration and activate α2 adrenoceptor. The present study was designed to determine whether α2 adrenoceptor activation and inhibition of neutrophil infiltration by Ber are involved in the improvement of LPS-induced cardiac dysfunction. METHODS: The mice were randomly divided into control, LPS, Ber+LPS, Ber+yohimbine (α2 adrenoceptor antagonist)+LPS, yohimbine+LPS, Ber, Ber+yohimbine and yohimbine groups. Water, Ber (50 mg/kg), Ber+yohimbine (50 mg/kg+2 mg/kg) or yohimbine (2 mg/kg) was given intragastrically once a day for 3 days. Normal saline or LPS (20 mg/kg) was injected intraperitoneally 1 h after intragastrical treatment on day 3. 12 h after LPS injection, the cardiac functions were determined by the technique of high-resolution ultrasonography. The histopathological changes of the myocardium were observed under microscope. Furthermore, the myocardial myeloperoxidase (MPO) was determined by Western blotting. RESULTS: Pretreatment with Ber, Ber combined with yohimbine or yohimbine attenuated histopathological changes in the heart of LPS-challenged mice. Echocardiography evaluation demonstrated that LPS decreased the cardiac output and stroke volume at 12 h after LPS challenge, which were reversed by pretreatment with Ber, Ber combined with yohimbine or yohimbine significantly. Compared to control group, LPS increased the level of myocardial MPO. Pretreatment with Ber and Ber combined with yohimbine, but not yohimbine alone significantly reduced the amount of myocardial MPO. CONCLUSION: The improvement of cardiac dysfunction by Ber is independent of α2 adrenoceptor activation and neutrophil infiltration inhibition in endotoxemic mice. The activation of α2 adrenoceptor by Ber may contribute to the pathogenesis of LPS-induced cardiac dysfunction in mice.     

Effects of propofol on expression of AIF and cell apoptosis in brain tissues of the rats with LPS-induced brain injury

AIM: To investigate the effects of propofol on the expression of apoptosis-inducing factor (AIF) and cell apoptosis in brain tissues of rats with lipopolysaccharide(LPS)-induced brain injury. METHODS: Seventy-two male and female SD rats weighing 220~250 g were randomly divided into 3 groups (n=24 each). Cerebral edema was induced by injection of LPS at 1 mg/kg via left internal carotid artery in LPS group and LPS+propofol group. In control group, equal volume of normal saline was administered instead of LPS. The rats in LPS+propofol group received intraperitoneal injection of propofol at 100 mg/kg immediately after LPS administration. Six rats in each group were decapitated 6 h, 12 h, 24 h or 48 h after operation and the frontal lobe cortex were immediately removed for determination of the water content. The apoptotic neurons were detected by Annexin V-PI staining. The protein levels of AIF, NF-κB and caspase-3 were measured by immunohistochemistry. The protein expression of AIF was detected by Western blotting analysis. RESULTS: Compared with control group, the brain water content, the number of neuronal apoptosis and the protein expression levels of AIF, NF-κB and caspase-3 were significantly increased in LPS group and LPS+propofol group. Compared with LPS group, the results mentioned above were markedly reduced in LPS+propofol group. CONCLUSION: Propofol attenuates LPS-induced brain injury by decreasing AIF protein expression and inhibiting apoptosis.     

Effect of berberine and yohimbine on impaired enterocyte proliferation and intestinal injury in LPS-challenged mice

AIM: To evaluate the effects of oral berberine (Ber) and yohimbine (Y) in preventing intestinal damage and impaired enterocyte proliferation caused by lipopolysaccharide (LPS) in mice. METHODS: Male BALB/c mice were randomly divided into 8 groups: control, LPS, Ber+LPS, Ber+Y+LPS, Y+LPS, Ber, Ber+Y and Y. The mice were administered intragastrically with distilled water (0.1 mL/10 g), Ber (50 mg/kg), Ber (50 mg/kg) in combination with Y (2 mg/kg) or Y (2 mg/kg) once a day for 3 days. One hour after intragastrical treatment on the third day, LPS (18 mg/kg) or normal saline was injected intraperitoneally. Twenty hours after LPS administration, the histological changes of the intestine were observed, and injury score was assessed. The mucosal weight, villus height and the content of diamine oxidase (DAO) in the ileum were also measured. Furthermore, the proliferation of enterocyte was identified by immunohistochemical staining for proliferating cell nuclear antigen (PCNA). RESULTS: Compared to the control mice, the mice challenged with LPS resulted in intestinal injury, including significantly increased injury score, decreased gut mucosa weight, villus height and the DAO contents in ileum. Furthermore, enterocyte proliferation was inhibited significantly 12 h after LPS challenge. Pretreatment with Ber or Ber+Y significantly inhibited the intestinal injury, and attenuated the impairment of enterocyte proliferation induced by LPS. However, no significant difference in the above parameters between Ber+LPS group and Ber+Y+LPS group was observed. Treatment with Y only did not prevent LPS-induced intestinal injury. CONCLUSION: Pretreatment with berberine remarkably reduces LPS-induced intestinal injury and the impairment of enterocyte proliferation in an alpha 2 adrenoceptor-independent manner.     

Effects of PDS on rat brain cortical nuclear factor kappa B in LPS shock

AIM:To explore the molecular mechanism of brain tissue injury induced by lipopolysaccharide (LPS),the effects of panaxadiol (PDS) on the expression of nuclear factor kappa B (NF-κB) in cerebral cortex of rat with LPS shock were studied. METHODS:Rats were randomly divided into LPS roup,LPS+dexamethasone group,LPS+PDS group and control group. The DNA binding activity and protein expression of NF-κB were observed. These indices were assayed at 1 h and 4 h after intravenous injection of LPS (4 mg·kg^-1). RESULTS:EMSA showed that PDS inhibited NF-κB DNA-binding activity in nuclear extracts at both 1 h and 4 h after LPS injection,compared with the LPS group (P<0.01). Western blotting showed that PDS down-regulated the expression of p65 and p50 protein in the nuclear extracts compared with the LPS group. However,the expression of p65 and p50 protein from cytosolic extracts did not show any significant change. CONCLUSION:PDS may alleviate brain injury by inhibiting NF-κB activation.     

Protective effect of dexmedetomidine-ulinastatin combination on lipopolysaccharide-induced acute lung injury in rats

AIM：To investigate the effects of dexmedetomidine-ulinastatin combination on acute lung injury induced by lipopolysaccharide (LPS) in rats. METHODS：Male Wistar rats were randomly divided into 5 groups: saline control group (NS group) was given saline (5 mL/kg, iv) alone; LPS group (L group) was given LPS (10 mg/kg, over 10 min); dexmedetomidine+LPS group (L+D group) was treated with the additional administration of dexmedetomidine (1 μg·kg -1·h -1) immediately after LPS injection; ulinastatin+LPS group (L+U group) was treated with the addi-tional administration of ulinastatin (50 000 U/kg, ip) immediately after LPS injection; dexmedetomidine+ulinastatin+LPS group (L+D+U group) received dexmedetomidine (1 μg·kg -1·h -1) and ulinastatin (50 000 U/kg) immediately after LPS injection. The animals were sacrificed at 6 h after LPS or NS administration. Partial pressure of arterial oxygen (PaO 2), pH and base excess (BE) were measured, and the lungs were removed for evaluation of histological characteristics and determining the concentrations of TNF-α, IL-1β, macrophage inflammatory protein 2 (MIP-2), malondialdehyde (MDA), nitric oxide (NO), prostaglandin E 2 (PGE 2) and myeloperoxidase (MPO) in lung tissues, lung wet/dry weight ratio (W/D), and albumin in brochoalveolar lavage fluid (BLAF). The pulmonary expression of nuclear factor kappa B (NF-κB) p65 was evaluated by Western blotting. RESULTS：Compared with NS group, PaO 2, pH and BE was lower in L group, which was increased by treatment with dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone. Compared with NS group, LPS induced marked lung histological injury, which was less pronounced in the animals treated with dexmedetomidine-ulinastatin combination but not dexmedetomidine or ulinastatin alone. The levels of IL-1β, IL-6, MIP-2, MDA, NO and PGE 2 in the lung tissues increased in L group compared with NS group, which were reduced by dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone. The MPO activity, MDA level and W/D increased in the lung tissues in L group compared with NS group, which was reduced by dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone. Compared with NS group, the albumin concentration in the BLAF increased, which was reduced by dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone. Compared with NS group, the expression of NF-κB p65 increased in L group, which was reduced by dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone.CONCLUSION：Dexmedetomidine-ulinastatin combination has a protective effect on LPS-induced acute lung injury in the rats.     

Peroxisome proliferator-activated receptor γ agonist attenuates ICAM-1 and CINC-1 expression in lungs of rats with acute lung injury

AIM：To investigate the effects of rosiglitazone (ROSI),an agonist of peroxisome proliferator-activated receptor γ (PPARγ),on the lung expression of intercellular adhesion molecule-1 (ICAM-1) and cytokine-induced neutrophil chemoattractant (CINC) in rats with acute lung injury.METHODS：Thirty-six male Wistar rats were randomly divided into six groups：control group,ROSI group,GW9662 (a PPARγ antagonist) group,lipopolysaccharide (LPS,6 mg/kg,iv) group,ROSI-LPS group (0.3 mg/kg ROSI iv 30 min prior to LPS) and GW9662-ROSI-LPS group (0.3 mg/kg GW9662,iv,20 min before ROSI).Four hours after LPS injection,wet/dry weight (W/D) ratio,myeloperoxidase (MPO) activity,malondialdehyde (MDA) and CINC-1 concentrations were assayed in the lung tissues.Immunohistochemical analysis of ICAM-1 expression was also studied.RESULTS：Pretreatment with ROSI significantly attenuated LPS-induced increases in W/D ratio,MPO activity,MDA and CINC-1 concentrations as well as ICAM-1 expression in the lung tissues.The specific PPARγ antagonist GW9662 antagonized the effects of ROSI.CONCLUSION：Pretreatment with ROSI reduces LPS-induced lung injury in rats.The mechanism involves inhibition of the lung expression of ICAM-1 and CINC-1 by the activation of PPARγ.     

Effect of Xueshuantong injection on LPS-induced DIC in rabbits

AIM：To evaluate the effect of Xueshuantong injection on lipopolysaccharide (LPS)-induced experimental disseminated intravascular coagulation (DIC) in rabbits. METHODS：To establish the LPS-induced DIC model, LPS was continuously injected into the rabbit autricular vein for 6 h. The survival rate of the rabbits was recorded after 24 h. The plasma levels of alanine aminotransferase (ALT) and blood urine nitrogen (BUN) were detected. Activated partial thromboplastin time (APTT), prothrombin time (PT), platelet count and fibrinogen concentration were measured. The activity of protein C, antithrombin Ⅲ (ATⅢ) and the concentration of tumor necrosis factor α (TNF-α) were also determined. RESULTS：The survival rate of DIC rabbits was only 26.7%. BUN and ALT increased obviously. APTT and PT became much longer, platelet count, fibrinogen, protein C and ATⅢ decreased obviously, and plasma TNF-α increased remarkably. The intravenous administration of Xueshuantong injection increased the survival rate of DIC rabbits up to 66.7% in a dose-dependent manner. Xueshuantong injection also decreased the plasma levels of BUN, ALT, APTT, PT and TNF-α significantly, while increased the levels of fibrinogen, platelet, protein C and ATⅢ in plasma. CONCLUSION： Xueshuantong injection has therapeutic effect on LPS-induced DIC in rabbits.     

Effect of Siduqing decoction,a Chinese medicine,on survival rate and multiple organ dysfunction in mice challenged with LPS

AIM: To investigate the effect of siduqing decoction, a Chinese medicine, on survival rate and multiple organ dysfunction in mice challenged with LPS. METHODS: Mice were administered intragastrically with Siduqing decoction or distilled water (0.2 ml/10 g) twice a day for 3 days, two hours after Chinese herbal medicine treatment on day 3, LPS or normal saline was injected intraperitoneally, and survival rates in each group were recorded at 12-h intervals. In another experiment, mice were sacrificed at 12 h after LPS, lung, liver, kidney and small intestine were collected and processed for the H & E staining. In addition, Blood was collected at 10 h after LPS injection for determining alanine aminotransferase (ALT) activity, blood urea nitrogen (BUN) and creatinine (Cr) contents. RESULTS: At 96 h after LPS injection, the survival rate (27%, n=34) was lower in LPS group than Siduqing treatment group (65%, n=31, P<0.05). ALT activity, BUN and Cr contents in serum were higher in LPS group than control group, Siduqing treatment significantly attenuated a increase in ALT activity, BUN and Cr content in serum induced by LPS. Histological examination showed inflammatory injury in the lung and intestine, hemorrhage in the lung and kidney, degeneration, necrosis in the liver and kidney, while Siduqing treatment attenuated pathological changes induced by LPS. CONCLUSION: These data indicate that Siduqing has a protective effect against LPS-induced multiple organ injury and increases survival rate in mice challenged with LPS.     

The effect of nucleolin on interleukin-1β secretion induced by lipopolysaccharide

AIM: To explore the expression of nucleolin in lipopolysaccharide(LPS)-mediated inflammatory models, and further investigate the role of nucleolin in expression and secretion of LPS-induced interleukin-1β(IL-1β). METHODS: To establish inflammatory models, mice suffered intraperitoneal injection of LPS(15 mg/kg)and RAW264.7 cells were treated with LPS(500 μg/L).Western blotting were applied to identify the expression of nucleolin in these inflammatory models. After over-expression of nucleolin by transient pcDNA3.1-C23 transfection and down-regulation by transient transfection of nucleolin antisense oligonucleotides, the secretion of IL-1β were examined by enzyme-linked immunosorbent assay (ELISA) in LPS-stimulated RAW264.7 cells. RESULTS: Westem blotting assays showed that the 110 kD nucleolin increased in RAW264.7 cells treated with LPS (500 μg/L) and the lung tissues of the mice treated with LPS (15 mg/kg), while the 80 kD component of nucleolin decreased. ELISA showed that LPS-induced IL-1β release in RAW264.7 cells transfected with pcDNA3.1-C23 was higher than that in pcDNA3.1 empty vector transfected cells. LPS-induced IL-1β release in RAW264.7 cells transfected with C23 antisense oligonucleotide was lower than that in normal cells and scramble oligonucleotide transfected cells. CONCLUSION: In LPS-mediated mouse endotoxemia model and LPS-mediated RAW264.7 cell inflammatory model, the expression of 110 kD nucleolin was up-regulated, but 80 kD nucleolin fragment decreased. Nucleolin promoted secretion of LPS-induced IL-1β.     

Mechanism of carbon monoxide protects rat intestine against LPS

AIM: To study the effect of exogenous carbon monoxide on rat intestine attacked by LPS. METHODS: The experimental rats were divided into 6 groups randomly: normal group, lipopolysaccharide (LPS, 5 mg/kg) group, CO inhalation (250 mL/M3) group, CO intraperitoneal injection (2 mL/kg) group, LPS (LPS 5 mg/kg) with CO inhalation (250 mL/M3) group and LPS (LPS 5 mg/kg) with CO intraperitoneal injection (2 mL/kg) group. The PaO₂, PaCO₂, SO₂ and COHb were monitored. Rat intestine malondialdehyde (MDA) with thiobarbitric acid (TBA) and superoxide dismutase (SOD) with hydroxylamine were detected and heme oxygenase-1 (HO-1) mRNA expression were checked by RT-PCR. RESULTS: Low concentration of CO inhalation (250 mL/M3) and CO intraperitoneal injection (2 mL/kg) did not cause the rat hypoxia. Compared to control group, exposure exogenous CO, the MDA content in LPS attacked rat intestine decreased, the SOD activity and the expression of HO-1 mRNA increased. CONCLUSIONS: Low concentration of CO inhalation (250 mL/M3) and low dose CO intraperitoneal injection (2 mL/kg) are safe to rat. Exposure exogenous CO protects rat intestine against LPS attack, induces the HO-1 mRNA expression and exerts anti-inflammation via endogenous CO.     

Mechanisms for protection of berberine against LPS-induced acute lung injury in mice

AIM:To investigate the mechanisms by which berberine attenuates LPS-induced acute lung injury, and provide a new strategy for the treatment of the lung injury due to LPS. METHODS:BALB/c mice were randomly assigned into three groups (control, LPS group, and berberine treatment group). Mice were administered intragastrically with distilled water (0.1 mL/10 g) or neutral sulfate berberine (50 mg/kg) once a day for 3 days, 1 h after intragastrical treatment on day 3, LPS (20 mg/kg) or normal saline was injected intraperitoneally (ip). All animals were sacrificed 12 h after LPS injection, the left lung tissue sections were prepared for histology analysis and the right lung were used to determine the ratio of wet to dry lung tissue weight (W/D). In another experiment, bronchoalveolar lavage fluid (BALF) was collected, and then the total protein content, and the amounts of white blood cells (WBC) and polymorphonuclear neutrophils (PMN) in BALF were determined. Furthermore, the phosphorylation of cytosolic phospholipase A₂ (cPLA₂) was detected with immunohistochemical analysis by using phospho-cPLA2(Ser505) antibody, and the contents of thromboxane B₂ (TXB₂) in BALF, malondialdehyde (MDA) in the lungs, and activity of superoxide dismutase (SOD) in lung tissues were also determined.RESULTS:LPS induced acute lung injury, activated cPLA₂, and increased TXB₂ content in the BALF and MDA level in the lung tissue. The pretreatment with berberine significantly attenuated lung injury, lung edema and protein leakage induced by intraperitoneal injection of LPS. The expression of phospho-cPLA₂ in the lung tissues and TXB₂ content in the BALF in the berberine treatment group were lower than those in LPS group (P<0.05). In addition, the content of MDA in the lung tissue was lower in the berberine treatment group than LPS group (P<0.05), but there was no significant difference in activity of lung SOD between the berberine treatment and LPS group (P>0.05). CONCLUSION:Pretreatment with berberine remarkably reduces the LPS-induced lung injury, which is, at least in part, through inhibiting phosphorylation of cPLA₂ and decreasing lipid peroxidation. These findings provide a new strategy for the prevention and treatment of LPS-induced acute lung injury.     

Losartan inhibits LPS-induced GFAP expression via AMPK activation in mouse hippocampus

AIM: To investigate the effects of losartan on lipopolysaccharide (LPS)-induced glial fibrillary acidic protein (GFAP) expression, and to determine whether adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) activation is involved in the mechanism.METHODS: Adult male KM mice were divided into control group, LPS model group, losartan treatment group, and losartan and Compound C co-treatment group. To establish a model of central nervous system inflammation, the mice received daily intracerebroventricular injection of LPS (24 μg/d) for 2 d. Daily losartan administration (0.5, 1 or 5 mg·kg^-1·d^-1, ip) initiated at 14 d prior to LPS injection. Compound C (10 mg/kg, ip), a selective AMPK inhibitor, started to be injected daily at 2 d prior to LPS injection. The hippocampal tissues in each group were isolated at 3 d after the last LPS injection, and then the protein levels of GFAP, AMPK, p-AMPK, mammalian target of rapamycin (mTOR) and p-mTOR were determined by Western blot.RESULTS: Twice LPS injections significantly increased the expression of GFAP in the hippocampus (P<0.01). Losartan inhibited LPS-induced GFAP expression in a concentration-dependent way, and losartan at 5 mg·kg^-1·d^-1 significantly inhibited GFAP expression and AMPK activation (P<0.05), but it had no obvious effect on mTOR activation. Furthermore, Compound C significantly reversed the effect of losartan treatment on LPS-induced GFAP expression and AMPK phosphorylation (P<0.05).CONCLUSION: Losartan inhibits LPS-induced GFAP expression in the mouse hippocampus, and AMPK activation but not mTOR, is involved in the mechanism.     

CO2 与养分交互作用对番茄幼苗养分利用的影响

以番茄(Lycopersicon esculentumMill.)‘合作906’为材料进行溶液培养试验,设2个因子:CO2和营养液浓度;CO2浓度设正常(360μL/L)和倍增(720μL/L)2个水平;营养液浓度设基本营养液(日本山崎番茄营养液),微量元素采用阿农营养液配方的1/2、1/4、1/8、1/164个水平,完全试验方案8个处理,3次重复。pH为6·0±0·2,3d更换1次营养液。移植到1·2L盆(2株/盒)中,植株在CO2生长箱(VS-3DMC)中培养,全天施放CO2,白天25℃,晚上15℃,光照为14h/d,光照强度11000lx,相对湿度60%。46d时收获,根、茎、叶经蒸馏水冲洗吸干水分后,放入纸袋105℃杀青,75…     

Effect of safflower injection on LPS-induced disseminated intravascular coagulation in rabbits

AIM：To study the effect of safflower injection on lipopolysaccharide (LPS)-induced disseminated intravascular coagulation (DIC) in rabbits. METHODS：The rabbit model of DIC was induced by continuous infusion of LPS at a dose of 100 μg·kg-1·h-1 for 6 h. The treatment with safflower injection and heparin were started simultaneously with LPS infusion through the contralateral marginal ear vein. The activated partial thromboplastin time (APTT), prothrombin time (PT), platelet count and fibrin(ogen) concentration were measured. The plasma levels of fibrin(ogen) degradation products (FDP), alanine aminotransferase (ALT) and blood urine nitrogen (BUN) were detected. The activity of protein C and antithrombin III (AT III) and the concentration of tumor necrosis factor α (TNF-α) were also determined. RESULTS：The gradual injury of hemostatic parameters was induced by continuous infusion of LPS. APTT, PT, BUN, ALT and plasma TNF-α were increased obviously. The platelet count, fibrinogen, FDP, protein C and AT III were decreased significantly. The intravenous administration of safflower injection attenuated the increases in the levels of APTT, PT, BUN, ALT, FDP and plasma TNF-α induced by LPS infusion, and improved the plasma levels of fibrinogen, platelet, protein C and AT III. CONCLUSION: Safflower injection protects against LPS-induced DIC in rabbits．     

Mechanisms for a decrease in mortality of LPS-challenged mice by rhynchophylline

AIM: To observe effect of rhynchophylline （Rhy） on mortality and organ injury in endotoxemic mice and further investigate the mechanisms of its actions. METHODS: Male mice were randomly assigned into control, LPS, Rhy +LPS and Rhy group, and injected subcutaneously with normal saline (0.05 mL/10 g), or rhynchophylline once a day for 3 d, 1 h after subcutaneously treatment on day 3, LPS (20 mg/kg) or normal saline was injected intraperitoneally. Survival rate was recorded every 12 h for 6 d. In another experiment, 12 h after LPS injection, the left lung and intestine tissue sections were prepared for histological analysis and the right lung were used to determine the ratio of wet to dry lung tissue weight (W/D),the serum was collected to detect the concentrations of alanine aminotransferase（ALT）, aspartate aminotransferase (AST ), bloodureanitrogen (BUN) and creatinine (Cr). In addition, the concentrations of tumor necrosis factor-α (TNF-α), interleukin-1β(IL-1β) and interleukin-10 (IL-10) in serum at 2 h after LPS challenge were detected by enzyme-linked immunosorbent assay. The concentration of NO in serum at 8 h was detected by enzymic method. The effect of Rhy on survival rate of mice subjected to cecal ligation and puncture (CLP) was also observed. RESULTS: Mortality of mice challenged with LPS alone was higher significantly than that in control at 24 h after LPS challenge, pretreated with Rhy at a dose of 8 or 16 mg/kg increased markedly the survival rate of LPS-challenged mice. However, Rhy at a dose of 8 mg/kg significantly increased mortality of mice subjected to CLP. In the histological analysis, severe inflammation was observed both in the lung and intestine tissues in the LPS group. LPS elevated lung W/D, the levels of ALT, AST, BUN, Cr, TNF-α, IL-1β, IL-10 and NO in serum. Pretreatment with Rhy had no obvious improvement in the lung and intestine tissue injury, and no significant depression in the lung W/D and the serum levels of ALT, AST, BUN, Cr, IL-1β, IL-10 and NO, but decreased the level of TNF-α in serum significantly in LPS -treated mice. CONCLUSION: Pretreatment with Rhy reduces the mortality in endotoxemic mice, but not decrease the mortality of mice challenged with CLP, at least in part, through inhibiting the synthesis and secretion of TNF-α.