非小细胞肺癌组织基质金属蛋白酶-3和组织金属蛋白酶抑制剂-3的表达

目的 探讨非小细胞肺癌(NSCLC)组织中基质金属蛋白酶-3(MMP-3)和组织金属蛋白酶抑制剂-3(TIMP-3)的表达及其意义.方法 采用免疫组化S-P法检测56例非小细胞肺癌组织和35例癌旁正常肺组织中MMP-3和TIMP-3的表达.结果 非小细胞肺癌组织中MMP-3的表达显著高于癌旁正常肺组织(P<0.05).非小细胞肺癌中MMP-3的表达,有淋巴结转移者高于无淋巴结转移者(P<0.05),Ⅲ+Ⅳ期高于I+Ⅱ期(P<0.05).非小细胞肺癌组织中TIMP-3的表达显著高于癌旁正常肺组织(P<0.05),非小细胞肺癌中TIMP-3的表达有淋巴结转移者低于无淋巴结转移者(P<0.05).结论 MMP-3和TIMP-3表达与非小细胞肺癌的侵袭、转移密切相关. Abstract： Objective To investigate the expression of MMP-3 and TIMP-3 in non-small cell lung cancer tissue and their significance. Methods The expression of MMP-3 and TIMP-3 in 56 lung carcinoma tissues and 56 para-cancerous normal lung tissues were detected by immuno histochemishy tecneque. Results The expression of MMP-3 in non-small cell lung cancer tissues was significantly higher than that in para-cancerous normal lung tissues (P<0.05).The expression of MMP-3 was significantly higher in non-small cell lung cancer with lymph node metastasis than that in lung carcinomas without lymph node metastasis (P<0.05),and was significantly higher in stage-Ⅲ and stage-IV tumours than that in stage-I and stage-Ⅱtumours (P<0.05).The expression of TIMP-3 in non-small cell lung cancer tissues was significantly higher than that in para-cancerous normal lung tissues (P<0.05).The expression of TIMP-3 was significantly lower in non-small cell lung cancer with lymph node metastasis than that in lung carcinomas without lymph node metastasis(P<0.05). Conclusions The expression of MMP-3 and TIMP-3 is closely correlated with invasion and metastasis of non-small cell lung cancer.     

Matrix metalloproteinase-8 inhibitors mitigate sepsis-induced myocardial injury in rats

Background Sepsis-induced myocardial injury （SIMI） is caused by a variety of mechanisms. The aim of the study is to investigate the effects of metalloproteinase-8 （MMP-8） on SIMI and its mechanisms in rats. Methods Forty male Sprague Dawley rats were randomly divided into four groups： MMP-8 inhibitor （M81）, dexamethasone （DEX）, sepsis, and sham groups. The sepsis model was established by cecal ligation and puncture （CLP）. Rats in the M81 group immediately received an intraperitoneal injection of M81 （0.1 mg/kg） after CLP. Rats in the DEX group immediately received an intraperitoneal （IP） injection of DEX （2 mg/kg）. Rats in the sepsis and sham groups received intraperitoneal injections of normal saline. Rats were sacrificed 12 hours after CLP. Paraffin sections were stained with hematoxylin and eosin to observe the myocardium. The myocardial ultrastructure was observed with transmission electron microscopy. MMP-8, tumor necrosis factor-Q （TNF-a）, and interleukin-113 （IL-113） were detected by immunohistochemistry. The expression of MMP-8 was measured by Western blotting. TNF-a and IL-113 levels in serum and myocardial tissue were determined by enzyme-linked immunosorbent assay. Results Compared with the sham group, the myocardium in the sepsis group was seriously injured. MMP-8, TNF-α and IL-1β expression was higher in the sepsis group than in the sham group, Treatment with M81 or DEX, however, attenuated sepsis induced histopathological changes in the heart, and was associated with significant reductions in serum and myocardial levels of TNF-a and IL-113 （P 〈0.05）. M81 significantly inhibited MMP-8 expression in myocardial tissue （P 〈0.05）. In addition, treatment with DEX was not associated with a change in myocardial levels of MMP-8 （P 〉0.05）. Conclusion MMP-8 inhibitor attenuated myocardial injury in septic rats, which might be related to reduced expression of TNF-α and IL-1β.     

Clinical study of inflammatory factors in sputum induced early after lung volume reduction surgery

Background The aim of this study was to prospectively study the changes in neutrophil elastase （NE）, fibroblast growth factor 9 （Fgf9）, matrix metalloproteinase-9 （MMP-9）, tissue inhibitor of metalloproteinase 1 （TIMP-1） in sputum induced during the early period after lung volume reduction surgery （LVRS）. Methods From April to October 2005, ten consecutive patients with chronic obstructive pulmonary disease （COPD） underwent LVRS. Ten non-small cell lung cancer patients （stage II-Illa） received Iobectomy as a control group. The induced sputum was collected from both groups at six different times （two weeks before operation and postoperatively at 1, 2, 4, 6 and 10 days）. The level of NE, Fgf9, MMP-9 and TIMP-1 were measured using enzyme-linked immunosorbent assay. Results The pulmonary function （FEV~%） and arterial blood gases （PaO2 and PaCO2） were significantly different belween the groups. There were no significant differences in age, ejection fraction （EF）, and operation duration, but hemoglobin in the LVRS group was statistically higher than in the controls. At certain times, there were significant differences in NE, MMP-9, TIMP-1 and MMP-9/TIMP-1 （P 〈0.05） but not in Fgf9 between the two groups. The levels of NE and TIMP-1 were maximal at 2 days postoperatively and that of MMP-9 and MMP-9/TIMP-1 at 4 days postoperatively in the LVRS group. In the control group, maximal levels of NE and TIMP-1 occurred at 2 days postoperatively and that of MMP-9 and MMP-9/TIMP-1 at 1 day postoperatively. Ten days after surgery, all values of the control group were not significantly different from the baseline. In the LVRS group, the levels were significantly different from the pre-operative values （P 〈0.05） apart from TIMP-1. Conclusion The levels of NE, MMP-9, TIMP-1 and MMP-9/TIMP-1 of the LVRS group were different from those of the control group. The time course of these chanties may be related to LVRS and the underlying process of COPD.     

Pathogenesis of cigarette smoke-induced chronic obstructive pulmonary disease and therapeutic effects of glucocorticoids and N-acetylcysteine in rats

Background T lymphocytes and matrix metalloproteinase (MMP) play an important role in the pathogenesis of chronic obstructive pulmonary disease (COPD). However, the details of the mechanisms involved are unclear. The aims of this study were to investigate the changes in interferon-γ (IFN-γ), interleukin-4 (IL-4), MMP-9, MMP-12 and tissue inhibitor of metalloproteinase-1 (TIMP-1 ) levels in a smoke-induced COPD rat model and the therapeutic effects of glucocorticoids and Nacetylcysteine.Methods Male Wistar rats were exposed to cigarette smoke for 3.5 months. Budesonide or Nacetylcysteine was given in the last month. Lung function was measured at the end of the study. IL-4 and IFN-γ levels were then determined in bronchoalveolar lavage fluid and lung tissue samples by enzyme-linked immunosorbent assay. The expression of MMP-9, MMP-12 and TIMP-1 mRNA in lung tissue was determined by RT-PCR.Results In comparison with the control group, rats exposed to smoke had a significant increase in IL-4 and MMP-12 levels and a significant decrease in IFN-γ levels. In addition, the IL-4/ IFN-γ ratio and MMP-12/TIMP-1 ratio were both higher. At the same time, the ratio of forced expiratory volume in 0.3 second to forced vital capacity (FEV0.3/FVC) and dynamic compliance (Cdyn) decreased and expiratory resistance (Re) increased. By measuring pulmonary mean linear intercept and mean alveolar numbers, obvious emphysematous changes were observed in the smoke exposed group.After treatment with budesonide, IL-4 and MMP-12 decreased and IFN-γ increased. The IL-4/IFN-γ ratio returned to normal, though the MMP-12/TIMP-1 ratio remained unchanged. FEV0 JFVC was significantly higher and Re was significantly lower than that in untreated smoke exposed rats. No significant differences were found in pulmonary mean linear intercept and mean alveolar numbers.After treatment with N-acetylcysteine, IFN-γ increased and the IL-4/IFN-y ratio decreased. The MMP-12/TIMP-1 ratio remained unchanged. Re and Cdyn both improved obviously. No significant differences were found in pulmonary mean linear intercept and mean alveolar numbers. Correlation analysis indicated that IL-4 levels in lung tissue correlated negatively with FEV0.3/FVC (r=-0.53, P=0.001 ), IFN-γ levels in lung tissue correlated negatively with Re ( r=-0.63, P=0. 000) andpositively with Cdyn ( r=0.44, P=0. 009), and that the IL-4/IFN-γ ratio correlated negatively with FEV0.3/FVC ( r=-0.44, P=0.010 ) and Cdyn ( r=-0.42, P=0. 015 ) and positively with Re ( r=0.58, P=0.000). Finally, MMP-12 correlated negatively with FEV0.3/FVC (r=-0.36, P=0.026).Conclusions Cigarette smoke exposure increases IL-4 levels and decreases IFN-γ levels. This may be the result of smoke-induced changes in lung function. Budesonide can mitigate the chanqes in IL-4 and IFN-γ levels induced by smoke exposure.N-acetylcysteine has no effect on IL-4, but increases IFN-γ levels and brings the IL4/ IFN-y ratio back to normal. Cigarette smoke can also promote MMP-12 gene expression and elevate the MMP-12/TIMP-1 ratio. This effect may play a role in smokeinduced emphysema. Budesonide and Nacetylcysteine do not alter the MMP-12/TIMP-1 ratio in this study when given in the late phase of smoke exposure.     

Duodenal-jejunal bypass surgery on type 2 diabetic rats reduces the expression of matrix metalloproteinase-9 and tissue inhibitor of matrix metalloproteinase-1 in the thoracic aorta

Background Bariatric surgery offers a productive resolution of type 2 diabetes mellitus （T2DM）.The development of T2DM vasculopathy is due to chronic inflammation,which increases matrix metalloproteinase-9 （MMP-9） and tissue inhibitor of matrix metalloproteinase-1 （TIMP-1） expression.This study sought to examine MMP-9 and TIMP-1 expression in the thoracic aorta after duodenal-jejunal bypass （DJB） surgery on a T2DM rat model induced by a high-fat diet and low dose streptozotocin （STZ）.Methods Twenty-one T2DM Wistar rats induced by high-fat diet and low dose STZ were randomly divided into DJB and sham duodenal-jejunal bypass （S-DJB） groups.Ten Wistar rats were fed a normal diet as a control.Recovery of gastrointestinal function post-operation and resumption of a normal diet completed the experiment.Body weight,blood glucose,blood lipid levels,and MMP-9 and TIMP-1 expression levels in aortic endothelial cells were measured throughout.Results DJB rats showed significant weight loss 2 weeks post-operation compared with S-DJB rats.After surgery,DJB rats showed significant improvement and steady glycemic control with improved insulin sensitivity and glucose tolerance.They also exhibited improved lipid metabolism with a decrease in fasting free fatty acids （FFAs） and triglycerides （all P ＜0.05）.Immunohistochemistry showed decreased MMP-9 and TIMP-1 expression 12 weeks after surgery （P ＜ 0.01）.Conclusions DJB surgery on an induced T2DM rat model improves blood glucose levels and lipids,following a high-fat diet and low dose STZ treatment.In addition,DJB decreased MMP-9 and TIMP-1 expression in vascular endothelial cells,which may play an important role in delaying the development of T2DM vascular disease.     

Effect of Dachengqi Decoction on NF-κB p65 Expression in Lung of Rats with Partial Intestinal Obstruction and the Underlying Mechanism

To investigate the effect of Dachengqi decoction on NF-κB p65 expression in lung of rats with partial intestinal obstruction and the underlying mechanism, 30 SD rats were randomly divided into three groups： sham-operation group, model group and Dachengqi decoction treatment group （Dachengqi group）, with 10 animals in each group. The models were made by partially ligating their large intestines outside the body. The pathological changes were analyzed by HE staining. The expression of NF-κB p65 in rats lung were measured by using real-time polymerase chain reaction and immunohistochemistry respectively. Moreover, the expression of caveolin-1 in rats lung was also measured to. Increased edema, interstitial thickening, hemorrhage, and infiltration of inflammatory cells were found in the model group. In contrast, this change was significantly reduced in Dachengqi group as compared with model group. In addition, the up-regulated caveolin-1 and NF-κB p65 were also suppressed by Dachengqi decoction in lung of rats with partial intestinal obstruction. We are led to concluded that the caveolin-l-NF-κB pathway plays an important role in the development of lung injury of rats with partial intestinal obstruction and Dachengqi decoction could down-regulate the expression of caveolin-1 and NF-κB p65 in lung of rats with partial intestinal obstruction.     

Expression of Leukemia Inhibitory Factor in Airway Epithelial Tissue of Asthmatic Rats

In order to investigate the expression of leukemia inhibitory factor(LIF) in airway epithelial tissues of normal and asthmatic rats,the influence of dexamethasone and the role of LIF in pathogenesis of asthma,30 Sprague-Dawley(SD) rats were randomly divided into 3 groups(10 for each group):normal group,asthma model group,and dexamethasone-interfered group.In asthma model group and dexamethasone-interfered group,asthma rat models were established by intraperi-toneal(i.p.) injection of 10% ovalbumin(OVA) and challenge with 1% OVA via inhalation.Rats in dexamethasone-interfered group were pretreated with dexamethasone(2 mg/kg,i.p) 30 min before each challenge.The expression of LIF protein in lung was detected by immunohistochemistry.The results showed that LIF protein was mainly expressed in cytoplasm of bronchial epithelial cells.The expression of LIF protein in the airway epithelial tissue of asthma model group was significantly higher than that in normal group and dexamethasone-interfered group(P<0.01),but there was no sig-nificant difference between normal group and dexamethasone-interfered group(P>0.05).It was con-cluded that the expression of LIF was increased significantly in the airway epithelial tissue of the asthma rats,and dexamethasone could down-regulate the expression of LIF.It was suggested that LIF might play an important role in the pathogenesis of asthma as an inflammation regulator.     

Heme oxygenase-1 expression in rats with acute lung rejection and implication

This study investigated the expression of hemeoxygenase-1 （HO-1） in rats with acute lung rejection and its implication. A valid rat orthotopic left lung transplantation model （SD rat→Wistar rat） was established by using an improved three-cuff anastomosis technique. The rats were divided into control group, CoPP （HO-1 inducer）-treated group and ZnPP （HO-1 inhibitor）-treated group. The severity of acute rejection was graded on the basis of the morphologic changes of the lung samples stained with HE. The expression of HO-1 protein in lung tissue was detected by using immunohistochemistry and Western blot, and HO-1 mRNA activity was assayed by RT-PCR. The results showed that the expression of HO-1 protein was significantly increased with the acute rejection grading in rats （P〈0.01）. As compared with control and ZnPP-treated groups, the severity of acute rejection was not alleviated and the grade not reduced significantly in CoPP-treated group （P〉0.05）. It was concluded that HO-1 protein might be involved in the pathological process of post-graft acute rejection. The expression of HO-1 protein was increased gradually with aggravation of acute rejection, and HO-1 protein might be used as an index to monitor acute rejection after lung transplantation.     

Synergistic Anti-inflammatory Effect of Radix Platycodon in Combination with Herbs for Cleaning-heat and Detoxification and Its Mechanism

Objective:To investigate the synergistic anti-inflammatory effect of Radix Platycodon in combination with herbs for cleaning-heat and detoxification and its mechanism for Fei(肺)-targeting.Methods: Forty Wistar rats were randomly divided into five groups(8 per group):the sham-operated group,model group, Radix Platycodon group,Flos Lonicera and Fructus Forsythia(LF) group,and Radix Platycodon,Flos Lonicera and Fructus Forsythia combination(PLF) group,using a random number table.A rat chronic obstructive pulmonary disease(COPD) model was established by passive smoking and intratracheal instillation of lipopolysaccharide(LPS).The treatments started from the 15th day of passive smoking for a total duration of 14 days.At the end of the treatment,changes in the following measurements were determined:lung histopathology, inflammatory cytokines including tumor necrosis factorα(TNF-α),transforming growth factorβ(TGF-β) and interleukin IL-1β(IL-1β) in bronchoalveolar lavage fluid(BALF),and mRNA expression of endogenous active substance intestinal trefoil factor 3(TFF3) in the lung tissue.Results:Light microscopy showed that compared with the sham-operated group,rats in the COPD model group had disrupted alveolar structure,collapsed local alveoli,significantly widened or even fused alveolar septa,and massive infiltration of inflammatory cells in the alveolar wall and interstitium.In addition,significant bronchial epithelium hyperplasia,partially shed epithelia, and marked inflammatory cell infiltration in the bronchial wall and its surrounding tissues were noticed.Electron microscopy showed that rats in the model group had degeneration of alveolar type II epithelial cell;reduction, breakage or even loss of cell surface microvilli;swollen mitochondria with disappearing cristae and vacuole-like structure;and,increased secondary lysosomes in alveolar macrophages.The TNF-α,TGF-βand IL-1βlevels and white blood cell(WBC) count in BALF were significantly increased(P<0.01 or P<0.05) and TFF3 mRNA expression in the lung tissue was significantly reduced(P<0.01).After treatment,the pathological morphology of lung injury was less severe in all three treatment groups.In addition,TGF-βand IL-1βand WBC count in BALF were decreased(P<0.01 or P<0.05),and TFF3 mRNA expression in the lung tissue was significantly increased in the PLF group(P<0.01).Compared with the LF group,the IL-1 p in BALF was significantly decreased (P<0.05),and TFF3 mRNA expression was significantly increased(P<0.05) in the PLF group.Conclusions:Radix Platycodon synergizes with herbs for cleaning-heat and detoxification in reducing inflammatory injury in a rat model of COPD.The synergistic anti-inflammatory effect is reflected in the improvement in pathological changes and in the reduction of IL-1 p levels in BALF.The mechanism of such synergistic action may be related to its effect on maintaining the TFF3 mRNA expression and Fei-targeting function.     

Expression of Lung Surfactant Proteins SP-B and SP-C and Their Modulating Factors in Fetal Lung of FGR Rats

This study investigated the expression of lung surfactant proteins SP-B and SP-C, and their modulating factors TTF-1 and PLAGL2 in the fetal lung of rats with fetal growth restriction(FGR). The rat FGR model was established by prenatal hypoxia in the first stage of pregnancy, 180 rats for experiment served as hypoxia group, and 197 healthy rats served as normal control group. The FGR incidence in hypoxia was compared with that in normal control group. The histological changes in the fetal lung were observed under the light microscope and electronic microscope in two groups. The SP-B, SP-C, TTF-1 and PLAGL2 proteins were determined in the fetal lung of two groups immunohistochemically. The expression levels of SP-B, SP-C, TTF-1 and PLAGL2 protein and m RNA in the fetal lung of two groups were detected by using Western blotting and RT-PCR respectively. The FGR rat model was successfully established by using hypoxia. Pathologically the fetal lung developed slowly, and the expression levels of SP-B, SP-C, TTF-1 and PLAGL2 protein and mR NA in the fetal lung were significantly reduced in hypoxia group as compared with those in normal control group. It was suggested that maternal hypoxia in the first stage of pregnancy could induce FGR, and reduce the expression of SP-B and SP-C, resulting in the disorder of fetal lung development and maturation.     

Expression of cyclooxygenase-2 and its pathogenic effects in nonalcoholic fatty liver disease

Objective:To investigate the expression of cyclooxygenase-2 and its pathological effect in the experimental nonalcoholic fatty liver of rats,and to explore its possible mechanism.Methods:The rat NAFLD model was established by giving a fat-enriched diet.The blood samples were obtained form abdominal aorta and the levels of serum ALT,AST and IL-1,changes in the hepatic tissue 6-k-PGF1α TXB2 were measured.The expression level of COX-2 in rats livers were assayed by immunohistochemistry,RT-PCR and Western-blot.Results:Light microscope analysis revealed that hepatocytes were injured in the model group and slightly in the treatment group.The levels of serum TXB2 and IL-1 in the fatty liver rats were increased.Compared with the model group,the IL-1 and TXB2 increased significantly(P < 0.05),on the contrary,compared with the normal group,the hepatic tissue 6-Keto-prostagland decreased significantly in the model group(P < 0.05),the treatment group also increased but P > 0.05.There was no positive expression of COX-2 in hepatic tissue of normal rats.In the model group,there was positive expression of COX-2 antigen and the number of COX positive cells progressively increased at 4,8,12 wks.The intensity of expression of COX-2 had significantly increased(P < 0.05)and the intensity of COX-2 expression in the treated group decreased remarkably compared with the model group(P < 0.05).The expression of COX-2 mRNA and the level of COX-2 protein were significantly stronger in the liver of model rats compared with normal rats,and significantly weaker in treated rats,than in 8W and 12W model rats(P < 0.05).Conclusion:The increase of COX-2 expression in NAFLD is closely associated with the severity of liver inflammation and damage.COX-2 may play an important role in the progression of rat NAFLD,and the expression of COX-2 mRNA is downregulated by cyclooxygenase-2 inhibitor,which can depress the oxidative stress and control inflammatory response efficiently.     

The role of angiopoietin-1 and thrombospondin-1 in the kidney of rats subject to 5/6 nephrectomy

The expression of angiopoietin- 1 （Ang- 1） and thrombospondin- 1 （TSP- 1） in 5/6 subtotal nephrectomy （STN） rats model, and its correlation to the renal microvasculature injury were investigated. Rat 5/6 STN model was established in adult male SD rats, and the sham-operated group and 5/6 STN group were set up. The renal function and histopathological changes were examined at the 1st, 2nd, 4th, 8th and 12th week after operation. The expression orAng-1, TSP-1 and CD31 in renal tissues was detected by using immunohistochemistry. From 2nd to 8th week after operation, Ang-1 was significantly expressed in glomeruli of rats with STN. Ang-1 staining in glomeruli of STN group was increased significantly as compared with that in sham-operated group at 4th and 8th week after operation, and subsequently decreased after the 12th week. The expression of TSP-1 was increased significantly in STN group. As compared with sham-operated group, the CD31 expression was significantly down-regulated from the 2nd week. The expression of Ang-1 mRNA was detected by using RT-PCR at the same time points. The expression of Ang-1 mRNA in renal tissue of rats with STN was significantly up-regulated at the 2nd, 4th and 8th week after operation as compared with that in STN group at other time points or in sham-operated group at the same time points, while decreased evidently at the 12th week as compared with that in sham-operated group. It is concluded that there are changes in the mRNA expression of Ang-1, and the significant up-regulation of the expression of TSP-1 in renal tissue of rats with STN, which may be involved in the remnant renal microvasculature injury.     

Investigation of Matrix Metalloproteinase -1, 2 and tissue inhibitor of matrix metalloproteinases-1 in Endometriosis

Objective： To explore the role of matrix metalloproteinase-1,2 （MMP-1, MMP-2） and tissue inhibitor of matrix metalloproteinases-1 （TIMP-1） in endometriosis. Methods： The eutopic and ectopic endometria from 40 subjects suffering from endometriosis and regular.endometria from 40 subjects （excluding endometriosis） were collected and examined by in situ hybridization technology and western blot assay. Results： Both expressions of MMP-1 and -2 were stronger in ectopic endometrium and eutopic endometrium than in normal endometrium. On the contrary, the expression of TIMP-1 in ectopic endometrium and eutopic endometrium was lower. The differences were significant （P 〈 0.01 ）. Moreover, there was no relationship among the expressions of MMP-1, 2 and TIMP-1 in ectopic endometrium. Conclusion： The expressions of MMP-1, 2 and TIMP-1 lose balance and lack of periodic changes in ectopic endometrium , which explains the biological invasive behavior of endometriosis. It was suggested-that regulating the balance between the MMPs and TIMP-1 should be an ideal therapeutic target to endometriosis.     

Role of Nuclear Transcription Factor-κB in Endotoxin-induced Shock in Rats

To investigate the role of NF-κB in endotoxic shock in rats. the model of endotoxinshock rats was induced by intravenous infusion of lipopolysaccharidc (LPS). 1 h. 2 h. 4 h and 6 h after LPS injection, the activation of NF-κB in blood mononuclear cells and the content of TNF-α and IL-6 in plasma was detected by enzyme-linked immunoadsordent assay (ELISA). The level of mean arterial pressure (MAP) and the histopathological changes of lung and liver were also observed. The activation of NF-κB in mononuclear cells increased 1 h after LPS injection and reached its peak 2 h after the injection, and its level was higher than that of normal group. The level of TNF-α was increased 1 h after the infusion and peaked 2 h after the injection, and its level was higher than that of normal group after LPS infusion. The content of IL-6 increased gradually with time. the IL-6 level was higher than that of normal group after LPS injection. MAP was decreased gradually with time and its level was lower than that of normal group after LPS injection. Pathological examination showed that endotoxic shock could cause pulmonary alveolar hemorrhage, edema and infiltration of inflammatory cell in lung tissue and congestion, edema, capillary dilation and inflammatory cell infiltration in liver tissue. It is concluded that NF-κB can up-regulate the expression of TNF-α and IL-6 in plasma and play an important role in endotoxin induced shock in rats.     

Effects of pioglitazone on expressions of matrix metalloproteinases 2 and 9 in kidneys of diabetic rats

Background The changes in matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) expressions were examined in the kidneys of diabetic rats to investigate the degradative pathway of collagen type Ⅳ （C-Ⅳ） and the protective effects of pioglitazone on an experimental model of diabetic nephropathy.Methods In 54 SD rats used in our study, 18 served as normal controls. Diabetes mellitus was induced in 36 age- and weight-matched rats by intraperitoneal injection of streptozotocin (70 mg/kg); 18 of the diabetic rats were allocated at random to receive pioglitazone ［20 mg·kg（-1）·d（-1）］ in their drinking water and 18 served as diabetic controls. Rats were killed after 2, 4, or 8 weeks of treatment. Kidneys were examined pathomorphologically and the expressions of MMP-2, MMP-9, and C-Ⅳ were analyzed by immunohistochemistry, and the results were quantified by image analysis techniques.Results Diabetes mellitus was associated with a decrease in the expression of MMP-2 in the glomeruli (P&lt;0.05, vs control). By contrast, MMP-2 expression in the interstitium increased, but not significantly (P&gt;0.05, vs control). The expression of MMP-9 did not show any change when comparing the three groups (P&gt;0.05, vs control). STZ-diabetic rats were also associated with an increase in the expression of C-Ⅳ in the glomeruli and the interstitium (P&lt;0.05, vs control). All diabetes-associated changes in MMP-2 expression were attenuated by pioglitazone treatment in association with reduced C-Ⅳ accumulation.Conclusions These results indicate that a decrease in MMP-2 expression in the glomeruli of diabetic rats may lead to impairment of C-Ⅳ degradation and contribute to the matrix accumulation in diabetic nephropathy. Pioglitazone treatment, which can attenuate the decrease of glomerular MMP-2 and the increase of C-Ⅳ degradation, has curative effects on diabetic nephropathy.     

Effect of All-trans Retinoic Acid on Airway Inflammation in Asthmatic Rats and Its Mechanism

The inhibitive effects of all-trans retinoic acid (ARTA) on airway inflammation in asthmatic rats and its mechanism on the basis of the regulation of nuclear factor kappaB (NF-kB) were explored. Thirty-two SD rats were randomly divided into 4 groups: control group, asthma group,dexamethasone treatment group and retinotic acid treatment group. The total and differential cell counts in the collected bronchoalveolar lavage fluid (BALF) were measured. The pathological changes in lung tissues were estimated by scoring. The expression of NF-kB inhibitor (IkBa), NF-kB,intercellular adhering molecule-1 (ICAM-1) in lung tissue was detected by immunohistochemical method. The results showed that in the two treatment groups, the total cell counts and proportion of inflammatory cells in BALF were significantly reduced, but there was no significant difference in differential cell counts in BALF between them. The pathological changes in lung tissues in the treatment groups were significantly attenuated as compared with asthma group. Except the epithelial injury in retinotic acid treatment group was milder than in dexamethasone treatment group, the remaining lesions showed no significant difference between them. In the two treatment groups, the expression of IkBa was increased, while the expression of NF-kB and ICAM-1 decreased with the difference between the two groups being not significant. It was concluded that the similar anti-inflammatory effects and mechanism of ATRA on airway in asthmatic rats to those of dexamethasone were contributed to the increase of cytoplasmic IkBa content and suppression of NF-kB activationand expression.     

Effects of Bone Marrow-derived Mesenchymal Stem Cells on TGF-β and MCP-1 in Injured Lung of Rats

The primary objective of the present study is to investigate the therapeutic effect of bone marrow-derived mesenchymal stem cells （MSCs） on bleomycin （BLM）-induced lung injury of rats and the effect on transforming growth factor-β （TGF-β） and monocyte chemoattractant protein-1 （MCP-1）. MSCs were isolated from SD rats. The recipients rats were divided randomly into four groups： lung injury group, MSC treatment group, MSC control group and normal control group. Rats of lung injury group and MSC treatment group were perfused with BLM of 5mg/kg （0.2-0.3ml） intratracheally, others were perfused with normal saline. After twelve hours, rats of MSC treatment group and MSC control group were injected MSCs of 0.5×10^6per rat into tail vein. Haematoxylin-eosin staining was used to observe the morphology in lung tissue. ELISA was used to detect the contents of TGF-β and MCP-1 in serum and bronchoalveolar lavage fluid （BALF）. Collagen content of the lung tissue was assessed by hydroxyproline （HYP） concentration. It was found that the thickness of alveolar wall and lung interstitium were significantly reduced in the rats of MSC treatment group compared with the lung injury group. HYP content in lung interstitium, TGF-β and MCP-1 in serum and BALF were increased significantly in rats of lung injury group two weeks after BLM perfusion, but they were reduced significantly in the rats of MSC treatment group compared with the injured rats. These observations provide evidence that MSCs engraftment could alleviate bleomycin-induced lung injury and fibrosis in rats and the therapeutic effects might relate with the decrease of TGF-β and MCP-1.