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Background Studies have determined that nucleotide binding oligomerization domain 2 (NOD2) plays a key role in innate immune response.However,whether NOD2 participates in the nature defense of fungal keratitis is unclear.Objective This study was to investigate the expression and significance of NOD2 on cornea in the initial of Aspergillus fumigatus keratitis (AFK) in rats.Methods Seventy-two adult clean Wistar rats were randomized into the normal control group,only corneal epithelial scraped group and AFK model group,and the AFK models were established by incubating Aspergillus fumigatus to cornea after corneal epithelium was scraped.All the operations were performed in the right eyes of rats.Reverse transcription PCR (RT-PCR) was carried out to detect the expression of NOD2 mRNA in corneal epithelium 4,8,16,24 hours after operation.Twenty-four hours after operation,the expression of NOD2 protein in rat corneas was examined by immunochemistry and immnunofluorescence technology.Also,the rat corneas were obtained for regular histopathological examination.The use and care of the animals complied with Institutional Animal Care and Use Committee Guidebook by NIH.Results All the models were made successfully.RT-PCR revealed that a fewer NOD2 mRNA were expressed on cornea in the normal control group,but the expressing levels of NOD2 mRNA were increased in the only corneal epithelial scraped group and AFK model group.Compared with only corneal epithelial scraped group,the elevated values of NOD2 mRNA expression in the AFK model group were statistically significant at 4,8,16 and 24 hours after operation (t =-0.409,-0.439,-0.534,-0.618,all at P=0.000).The histopathological examination displayed that the cornel tissue had intact structure in the normal control group,and partly corneal epithelial deficiency,slight corneal swelling and fewer neutrophil granulocytes were seen in the only corneal epithelial scraped group.However,corneal ulcer,severe corneal edema and a lot of neutrophil granulocytes were exhibited in the AFK model group.Immunochemistry and immnunofluorescence staining evidenced that weaker expression of NOD2 was visualized in the corneal epithelial and endothelial layers,and obviously enhanced staining was seen in the AFK model group.The expressing levels (absorbancy) were 0.045 ± 0.005,0.050 ± 0.005 and 0.092 ± 0.006 in the normal control group,only corneal epithelial scraped group and AFK model group,respectively,showing a significant increase in the AFK model group compared with the only corneal epithelial scraped group (t =0.042,P =0.000).Conclusions Expression of NOD2 is upregulated in the corneas with AFK,suggesting that NOD2 participates the natural defense in the initial of fungal keratitis.NOD2 may play an important role in the process of anti-fungal innate immune response in cornea.
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AIM: To investigate the effect of phosphatidylinositol 3-kinase (PI3K) inhibitor wortmannin on expression of inducible nitric oxide synthase (iNOS) in bronchiole epithelial cells of asthma rat.METHODS: 24 male Sprague-Dawley rats were randomly divided into 3 group, namely control group, asthma group and PI3K inhibitor wortmannin+asthma group. The numbers of total cells and eosinophil cells in bronchoalveolar lavage fluid (BALF) were counted. The expression of inducible nitric oxide synthase was detected by immunohistochemistry method, and iNOS mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). The activity of PI3K, iNOS and content of NO in lung homogenates were tested by spectrophotometry.RESULTS: The numbers of eosinophils and the ratios of eosinophils to total cells in BALF in asthma group were significantly higher than those in control group, while those in wortmannin+asthma group were decreased compared to asthma group. The activity of PI3K, iNOS and content of NO in lung homogenats in asthma group were significantly higher than those in control group, while those in wortmannin+asthma group were decreased compared to asthma group. The expression of iNOS protein in bronchiole epithelial cells and mRNA in lung homogenates in asthma group were markedly increased compared to control group, while those in wortmannin+asthma group were decreased compared to asthma group.CONCLUSION: PI3K regulates the expression of iNOS in airway of asthma rat and affects inflammatory reaction in airway.
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Objective To investigate the influence of injection carthamus tinctorius D. (1C) on the expression of intercellular adhesion molecule-1(ICAM-1) during the ischemia-reperfusion injury of lung (LIRI) in rabbits and its potential mechanism. Method Single lung ischemia-reperfusion animal model was induced in rabbits. A total of 30 Japanese white rabbits were randomly divided into sham-operation group (S group, n =10), ischemia-reperfusion group (I/R group, re = 10) and ischemia-reperfusion plus 1C group (1C group, n = 10) .The rabbits of 1C group received 1C 2.0 ml/kg injected intravenously just at 20 min before ligation of artery involved and the same dose of 1C instantly at the initiation of reperfusion. Malondialdehyde (MDA) , superoxide dismutase (SOD) and xanthine oxidase(XO) in serum were measured. The lung tissue was sampled and assayed wet/dry weight ratio (W/D), contents of myeloperoxidase (MPO) at the end of the experiment, and ultrastructure changes were observed under electron microscope. The expression of ICAM-1 was measured by using immunohistochemistry(IHC) . snd one-way ANOVA was used for statistical analysis. Results In I/R group, serum XO and MDA increased and SOD decreased, whereas the same pattern of changes but less magnitude happened in 1C group ( P < 0.01). The values of W/D and MPO were much higher in I/R group, but lower in 1C group. Under electron microscope, the ultrastructure of lung tissue showed pathological changes in the rabbits of I/R group,and these changes were greatly attenuated in the rabbits of 1C group . The IHC showed that ICAM - 1 in lung tissue of I / R group was (2.94±0.48) which was significantly higher than that of 1C group(1.75 (P < 0.01). Conclusions Injection Carthamus tinctorius D. may meliorate the ischemia-reperfusion injury of lung by way of suppressing the expression of ICAM-1, inhibiting neutrophil aggregation, lowering oxygen free radical level and decreasing lipid peroxidation.
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Objective To investigate the change of caspase-3 in rabbits after lung ischemia-reperfusion injury(LIRI) and the effect of puerarin(葛根素).Methods Thirty healthy rabbits used for unilateral lung ischemia-reperfusion model were randomly divided into 3 groups(each n=10): control group(C group),lung ischemia-reperfusion group(I/R group) and puerarin group.The activity of serum superoxide dismutase(SOD),the contents of serum malondialdehyde(MDA) and nitric oxide(NO),the wet to dry weight(W/D) ratio of lung tissue and the index of quantitative assessment of histological lung injury(IQA) were measured respectively in different groups;the pneumocyte apoptosis index(AI) was achieved by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL);caspase-3 protein and mRNA expression were studied by using in situ hybridization(ISH) and immunocytochemistry(IHC) techniques in the groups mentioned above.Results The activity of SOD and content of NO were significantly lower in I/R group than those in C group(both P
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Objective To explore the relationship between the change of intestinal environment and pathogenesis of non alcoholic steatohepatitis (NASH). Methods Forty two SD rats were divided into 3 groups randomly: model group( n =24), treatment group( n =12) and normal group( n =6). The rats of model group and treatment group were given fat rich diet, and those of normal group were given normal diet. Furthermore, the rats of treatment group were given lactulose after 8 weeks of fat rich diet feeding. Twelve rats of model group were sacrificed at 8 weeks of study. At 16 weeks, the rats of treatment group, normal group, and the rest of model group were sacrificed. The serum levels of aminotransferase were measured and the pathology of livers were observed by HE stain. Results The rats' livers presented the pathology of steatohepatitis with higher serum levels of ALT and AST in the 16 weeks model group. The serum levels of ALT and AST of treatment group decreased significantly and were close to normal group. The hepatic inflammation scores also decreased markedly (5.83?2.02 vs. 3.63?0.64), but were still higher (3.63?0.64 vs. 1.98 ?0.90) than those of 8 weeks model group. And the degree of hepatocyte steatosis did not change in the treatment group. Conclusions Lactulose could ameliorate the hepatic inflammation of rats with steatohepatitis induced by fat rich diet, but could not prevent the development of steatohepatitis. It suggested the change of intestinal environment, such as intestinal bacteria overgrowth, was one of important factors in the pathogenesis of NASH.
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AIM:To study testicular pathological change and its pathogenesis. METHODS:Testicular structure of alloxan induced diabetic rats was observed under light microscopy (LM)and transmission electron microscopy(TEM). Activity of superoxide dismutase(SOD),glutathione peroxidase(GSH-PX)，nitric oxide synthase(NOS)and content of malondialdehyde (MDA),nitric oxide(NO) were detected biochemically in testicular homogenate. RESULTS:It is manifestated as atrophy of seminiferous tubule and germinal arrest under LM and expansion of mitochondria and smooth endoplasmic reticulum, cytoplasmic inclusion of sertoli cell under TEM.Activity of SOD, GSH-PX decreased while activity of NOS, content of MDA, NO increased in diabetic rats compared with control one. CONCLUSION:Disturbance of spermatogenesis and damage of sertoli cell are the main morphological change of diabetic testis, lipid peroxidation and NO may be involved in it.
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Aim To investigate the effect of ligustrazine on pulmonary arterial pressure,thromboxane A2(TXA2) and prostacyclin(PGI2) in chronic hypoxic hypercapnic rats.Methods Thirty rats were randomly divided into control group(A), hypoxic hypercapnic group(B) and hypoxic hypercapnia+ ligustrazine (lig) group (C) .Results The mPAP in group B was significantly higher than that in group A(P0.05) ; The specific value of WA/TA(vessel wall area/total area) and SMC (the density of medial smooth muscle cells) were significantly higher in group B than that in group A(P0.05).Conclusion Ligustrazine can inhibit pulmonary hypertension and structural remodeling of pulmonary artery in chronic hypoxic hypercapnic rats by inhibiting synthesis of TXA2.
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AIM: To investigate the effect of puerarin (Pur) on expression of Fas/FasL mRNA in lung tissue during pulmonary ischemia and reperfusion injury(PIRI) in rabbits.METHODS: Single lung ischemia and reperfusion animal model was used.The rabbits were randomly divided into three groups,sham operated group(sham,n=10),PIR group(I-R,n=30) and PIR+ Pur group(Pur,n=30).Changes of several parameters included apoptotic index(AI),wet to dry ratio of lung tissue weight(W/D) and index of quantitative assessment of histologic lung injury(IQA) were measured at 60,180 and 300 minutes after reperfusion in lung tissue.Meanwhile,the location and expression of Fas/FasL mRNA were observed.Lung tissue was prepared for light microscopic and electron microscopic observation at 60,180,300 minutes after reperfusion.RESULTS: As compared with group I-R,Fas/FasL mRNA slightly expressed in intima and extima of small pulmonary artery,alveoli,and bronchiole epithelia in group Pur.The values of AI,W/D and IQA showed significantly lower than that in group I-R at 60,180,300 minutes after reperfusion in lung tissue(P
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AIM: To study the role and the mechani sm of heme oxygenas/endogenous carbon monoxide on nitric oxide synthase/nitric oxide system in rats with pulmo nary hypertension induced by hypoxic hypercapnia. METHODS: Spr ague-Dawley rats w ere randomly divided into three groups: control group (A group),hypoxic hypercap n ic group (B group), hypoxic hypercapnia+hemin group (C group). Blood CO concentr at ion (COHb%),NO concentration,HO-1 activity, iNOS, cNOS in blood serum and lung h omogenate were measured, respectively. RESULTS: ① mPAP and RV /(LV+S) of B g roup were significantly higher than those of A and C group( P
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AIM: To clarify the role of nitric oxide (NO) system in development of chronic hypoxic hypercapnic pulmonary hepertension. METHODS: Male Sprague-Dawley rats were randomly divided into control group and hypoxic hypercapnic group. NO content of plasma was determined, constitutive nitric oxide synthase (cNOS) and inducible nitric oxide synthase (iNOS) were examined using the technique of immunohistochemistry, expression of cNOS mRNA and iNOS mRNA of arteriole were detected by in situ hybridization. RESULTS: Plasma NO concentration, cNOS activity and cNOS mRNA expression in arteriole of chronic hypoxic hypecapnic group were significantly lower than that of control group ( P
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AIM: To study the effect of L-arginine (L-Arg) on function and structure of mitochondria in ischemia-reperfusion (MRI) myocardial cells. METHODS: Thirty rabbits were randomly divided into three groups (n=10 in each), control group, MIR group and MIR+L-Arg group. The mitochondrial respiratory function, Ca~(2+) concentration ([Ca~(2+)]m), malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were determined. Meanwhile, the contents of adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), total adenylic acid number (TAN) and energy charge (EC) in the myocardial tissue were respectively measured. Moreover, the ultrastructure changes in myocardial mitochondria were observed during MIR. RESULTS: The mitochondrial respiratory control rate (RCR), velocity 3 (V_3), SOD, surface density (Sv) and specific surface (?) in MIR+L-Arg group were higher than those in MIR group, velocity 4 (V_4), [Ca~(2+)]m, MDA, volume density (Vv), horizental diameter (Hd) were lower than those in MIR group. ATP, ADP, TAN and EC levels of myocardial tissue were higher than those in MIR group. There was no significant difference between MIR+L-Arg and control group in V_3, V_4, SOD, MDA, Vv, Sv, ?, Nv, Vd, AMP and TNA. CONCLUSION: It is suggested that L-Arg improves the function and structure of mitochondria in myocardial cells in the reperfusion injury after myocardial ischemia by decreasing oxygen free radical level and Ca~(2+) overload in the mitochondria. [
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AIM: To investigate the variations of heme oxygenase-1/carbon monoxide system in lung ischemia-reperfusion injury and effects of puerarin on the system.METHODS: The unilateral lung ischemia-reperfusion model was replicated in vivo.Rabbits were randomly divided into three groups(n=10 in each): control group,ischemia-reperfusion(I-R) group and puerarin group.The blood specimens collected at times before ischemia,ischemia 1 h,reperfusion 1 h,3 h and 5 h were tested for the contents of carboxyhemoglobin(COHb) and cyclic guanosine monophosphate(cGMP).The lung tissues sampled at 5 h after reperfusion were assayed for wet/dry weight ratio(W/T),the injured alveoli rate(IAR) and the activity of HO-1.The changes of ultrastructure were observed under electron microscope.The tissue slides were also stained by immunohistochemistry(IHC) and in situ hybridization(ISH) to detect the expression of HO-1.RESULTS: The plasma content of COHb and cGMP in I-R and puerarin group increased in a time-dependent manner after I-R compared with control group,but the increment in puerarin group was higher than that in I-R group(P
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AIM: AIM: To explore the relationship between apoptosis in the lung tissues and lung ischemia/reperfusion injury, and observe effects of panax notoginseng saponins (PNS) on apoptosis in lung ischemia/reperfusion injury. METHODS: Single lung in situ ischemia/reperfusion animal model was used. Eighty four Japanese white rabbits were randomly divided into control group (control), ischemia/reperfusion 1 h group (IR1h), IR3h, IR5h, Panax Notoginseng Saponins 1 h group (PNS1h), PNS3h and PNS5h. TUNEL, immunocytochemistry and in situ hybridization techniques were used to observe apoptosis and Fas/FasL expression in various phases of lung ischemia/reperfusion. RESULTS: Cell apoptosis in lung tissues were significantly high, Fas/FasL mRNA and its protein were up-regulated in lung tissues of lung ischemia/reperfusion injury compared with control (all of P
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AIM: To investigate the effect of heme oxygenase-1 (HO-1)/carbon monoxide (CO) system on pulmonary ischemia-reperfusion injury (PIRI) in rabbits. METHODS: Single lung ischemia and reperfusion animal model was used in vivo. The rabbits were randomly divided into three groups (n=10 in each), control group (C), PIR group (I-R), PIR+ hemin group (H) and PIR+zinc protoporphyrin IX (ZnPP) group (Z). Changes of several parameters which included plasma carboxyhemoglobin (COHb) at different time points, wet to dry ratio of lung tissue weight (W/D), the injured alveoli rate (IAR) and the HO-1 enzymatic activity were measured at 180 min after reperfusion in lung tissue. The tissue slides were also stained by immunohistochemistry (IHC) and in situ hybridization (ISH) for HO-1 to detect the expression of HO-1 in lung and to analyze the optical density. The lung tissue was prepared for electron microscopic observation at 180 min after reperfusion. RESULTS: The plasma content of COHb in I-R, H, and Z group increased in a time-dependent manner after I-R. But the increment of H group was higher than that of I-R group, while that of Z group was lower. The HO-1 activity in lung tissue was highest in H group, followed by IR group, Z group, and C group (P
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AIM: To investigate the expression of soluble guanylate cyclase protein and its mRNA in rat pulmonary artery after exposure to hypoxia and hypercapnia. METHODS: Male Sprague-Dawley rats were randomly split into 4 group, which were hypoxic hypercapnic (HH 1 week, HH 2 weeks, HH 4 weeks) group and control group, to copy pulmonary hypertensive animal model. The expression of sGC? 1 and ? 1 subunits protein of medial and small pulmonary artery was performed by immunohistochemistry with a polycolonal antibody. In situ hybridization was performed on the rat lung tissue using sGC oligonuclear probe to assay the expression of sGC? 1 subunit mRNA. RESULTS: The sGC? 1 and ? 1 subunits protein and sGC? 1 subunit mRNA were faint staining in the pulmonary small and medium artery in HH1 week, HH 2 weeks and HH 4 weeks groups compared with control group (all P
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AIM: To investigate the effect of diltiazem on mean pulmonary arterial pressure(mPAP) and nitric oxide synthase(NOS) in arterioles in chronic hypoxic hypercapnic rats. METHODS: Twenty-four rats were randomly divided into three groups: control group(A),hypoxic hypercapnic group(B), hypoxic hypercapnia+ diltiazem group (C), constitutive endothelial NOS(ceNOS) were observed in arterioles of rats using the technique of immunohistochemistry,ceNOS mRNA were observed by the technique of in situ hybridization . RESULTS: (1)mPAP was significantly higher in rats of B group than that of A and C group( P 0 05),but mCAP was lower in rats of C group than that in B group.(2)Light microscopy showed WA/TA (vessel wall area/total area) was significantly lower in rats of C group than that of B group ( P
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AIM: To study the effect of chronic hypoxic hypercapnia on expression of heme oxygenase-1(HO-1). METHODS: Sprague-Dawley rats were randomly divided into three groups: control group(A),hypoxic hypercapnic group(B), hypoxic hypercapnia+hemin group(C). HO-1 and HO-1 mRNA were observed in pulmonary arterioles by the technique of immunohistochemistry and in situ hybridization. RESULTS: ① mPAP and weight ratio of right ventricle(RV) to left ventricle plus septum (LV+S) were significantly higher in rats of B group than those of A and C group (P0.05). ② Blood CO concentration was significantly higher in rats of B group than that of A group(P
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AIM: To study the protective effect of Ligustrazini(LGT) on gut barrier function after hemorrhagic shock-reperfusion. METHODS: Thirty white rabbits were divided randomly into 3 groups: control group (A),shock group (B) and LGT group (C). Malondialdehyde(MDA), tumor necrosis factor-?(TNF ?), interleukin-1?(IL-1?) and nitric oxide products(NO - 2/NO - 3) contents were measured in intestinal mucosa at 3 hours following reperfusion,culture of bacteria in blood from rabbits of 3 groups was carried out,the intestinal mucosa was examined under optical and electron microscope. RESULTS: MDA, TNF ?, IL-1? and NO - 2/NO - 3 contents of intestinal mucosa remained unchanged in group C,but increased significantly in group B, compared with group A. Incidence of bacterial translocation in group B was markedly higher than that in group A at 30 min following reperfusion,there was not any difference between group A and group C. Under light and electronic microscope,in comparison with A and C groups,intestinal mucosa damage in B group became more severe. CONCLUSION: LGT can protect gut barrier from intestinal ischemia-reperfusion injury induced by hemorrhagic shock through reducing oxygen free radicals,raising nitric oxide and preventing inflammation.
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AIM: To study testicular pathological change and its pathogenesis. METHODS:Testicular structure of alloxan induced diabetic rats was observed under light microscopy (LM)and transmission electron microscopy(TEM). Activity of superoxide dismutase(SOD),glutathione peroxidase(GSH-PX),nitric oxide synthase(NOS)and content of malondialdehyde (MDA),nitric oxide(NO) were detected biochemically in testicular homogenate. RESULTS: It is manifestated as atrophy of seminiferous tubule and germinal arrest under LM and expansion of mitochondria and smooth endoplasmic reticulum, cytoplasmic inclusion of sertoli cell under TEM.Activity of SOD, GSH-PX decreased while activity of NOS, content of MDA, NO increased in diabetic rats compared with control one. CONCLUSION: Disturbance of spermatogenesis and damage of sertoli cell are the main morphological change of diabetic testis, lipid peroxidation and NO may be involved in it.
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0.05) was observed. The protein expressions of TLR-4, NF-?B p65, HSP70 and ICAM-1mRNA in IR group were significantly increased as compared to C group and PD group, while those expressions in PD group were evidently higher than those in C group (all P