Effect of sevoflurane on tissue permeability of lung ischemia-reperfusion injury in rats

Objective:To investigate the effect of sevoflurane on tissue permeability of lung ischemiareperfusion injury(LIRI)in rats.Methods:A total of 45 wistar rats were randomly divided into3 groupsⅠ,Ⅱ,Ⅲ.Modified Eppinger method was adopted to establish the rat lung ischemiareperfusion injury model.GroupⅠserved as the control group,groupⅡas ischemia reperfusion group,groupⅢas sevoflurane ischemia-reperfusion group.Blood gas index,lung permeability index(LPI)change,lung tissue pathology change and lung water content were observed and compared between groups of rats at different time points.Results:During ischemia reperfusion,all rats kept balance of the MAP during different time points,SPO_2 of groupⅡandⅢdecreased significantly thanⅠgroup(P0.05);after reperfusion lung permeability index in GroupⅡandⅢwas higher than the control group significantly(P0.05),120 min after reperfusion LPI change and iujury of groupⅢwas significantly lower thanⅡgroup(P0.05);interstitial and alveolar cavity effusion in of groupⅢwere lower than that of groupⅡ.Conclusions:Sevoflurane pretreatment can reduce the lung tissue permeability,and LIRI plays a protective role in LIRI.     

Anti-tumor effect of matrine combined with cisplatin on rat models of cervical cancer

Objective:To observe the anti-tumor effect of matrine combined with cisplatin on U14 rat models of cervical cancer.Methods:A total of 80 female Kunming rats were used to establish U14 rat models of cervical cancer and then divided into groups Ⅰ,Ⅱ,Ⅲ and Ⅳ,with 20 rats in each.For Group Ⅰ,the control group,injection of normal saline was given around the tumors.For Group Ⅱ,injection of 2 mg/kg cisplatin was given around the tumors.For Group Ⅲ,injection of 75 mg/kg matrine was given around the tumors while the combined injection of matrine and cisplatin was given for Group Ⅳ with the same doses as Groups Ⅱand Ⅲ.The animals were sacrificed 10 d after the injection and tumors were taken out for the comparisons of tumor weights after injection and calculation of anti-tumor rates,while thymus and spleen were taken for thymus index and spleen index.Blood in eyeball was collected for determination of changes in serum creatinine and urea nitrogen levels.Sections of tumor issue were prepared and morphological changes in tumor tissue cells were observed by using immunohistochemistry technique.Results:After injection,the thymus index and spleen index in Groups Ⅲ and Ⅳ were significantly higher than those in Groups Ⅰ and Ⅱ(P0.05)while the two indexes in Group Ⅱ were significantly lower than Group Ⅰ(P0.05).The tumor weights in Groups Ⅱ and Ⅳ were significantly smaller than those in Groups Ⅰ and Ⅲ(P0.05) with significantly higher anti-tumor rates than Groups Ⅰ and Ⅲ(P0.05).The serum creatinine and urea nitrogen levels in Groups Ⅲ and Ⅳ were significantly lower than Group Ⅱ(P0.05) and the two indicators in Group Ⅲ were significantly lower than those in Group Ⅳ(P0.05).The observation under the histological microscope showed densely arranged tumor cells in Group Ⅰ,growing as a crumby structure and diffuse appearance,with hyperchromatic and large nuclei,and abundant cytoplasm.In the case of Group Ⅱ,it showed less tumor cells,with extensive degenerative necrosis,sparse arrangement and karyopyknosis as well as karyoclasis.For Group Ⅲ,necrosis of tumor cells in different sizes and heterogeneous color in nuclei were observed.For Group Ⅳ,the number of tumor cells was significantly smaller than Groups Ⅰ and Ⅲ and the tumor cells presented an appearance of crumby structure as cancer nests,with more proliferation of connective tissue.Conclusions:The treatment of matrine combined with cisplatin can significantly improve the anti-tumor effect on U14 rats with cervical cancer,which can be a new option for the treatment for cervical cancer.     

Therapeutic effect of Captopril on rheumatoid arthritis in rats

Objective:To investigate the therapeutic effect of the intervention treatment with different doses of Captopril on TNF-α contents in serum of rheumatoid arthritis(RA) rats,and to provide the theoretical proofs for clinical application of Captopril in treatments ol rheumatoid diseases.Methods:Fifty Wistar rats were randomly divided into 5 groups,namely.Group A,Group 13.Group C.Group D,Group E with 10 rats in each group.Injection of Freund's complete adjuvant was employed to establish adjuvant-induced arthritis model in rats.Group A was model group;after model establishment,rats were treated with 20 mL normal saline as placebo(ip.).Rats in Group B were treated with 8 mg/kg cyclophosphamide(ip.).Rats in Group C.D and E were intraperitoneally injected with 30 mg/kg.100 mg/kg and 300 mg/kg Captopril respectively.Rats in each group were subjected to continuous treatment for 3 weeks,and then sacrificed.Eyeballs of rats were excised and blood was collected.TNF- α content in serum were detected using ELISA:each group rats were compared for the hind legs arthrocele.Right ankle tissues of rats were collected to prepare section,and microscopic observation of pathological changes was performed.Results:TNF- α content in serum of Group A rats was significantly higher than that of rats in other 4 groups(P0.05).TNF- α content in serum of Group B rats was significantly lower compared with that of rats in Groups C.D and E.The highest TNF- α content in serum of rats treated with Captopril was found in Group C,followed by Groups D and E(P0.05).Right ankle arthrocele of rats in Groups B.C.D and E in early stage showed no statistical difference compared with that of Group A rats(P0.05).From Day 8,ankle arthrocele of rats in Groups B.C.D and E was obviously relieved compared with that of Group A rats:the anti-inflammatory effects were gradually enhanced with the extension of medication time.Treatments of Groups C.D and E showed significant activities against tardive aithrocele:the degree of ankle arthrocele in rats of these three groups was lower than that of Group A rats(P0.01).Histological observation showed that large amount of inflammatory cells and plasmocyte infiltration was found in ankle synovial tissues of Group A rats.Relief of hyperaemia and edema of right ankle synovial tissues as well as significant decrease in synoviocyte layer hyperplasia,intra—articular inflammatory cells infiltration and cartilage articularis damage degree etc.were observed in Groups B.C.D and E.Conclusions:Intervention treatment with Captopril can effectively reduce the TNF- α content in serum of rheumatoid arthritis rats and inhibit the generation of inflammatory factors,so as to achieve the therapeutic effect.     

Effect of abdominal trauma on hemorrhagic shock-induced acute lung injury in rats

AIM: To evaluate the effects of abdominal trauma on hemorrhagic shock-induced acute lung injury in rats. METHODS: Five groups were allocated (n = 8) in the study. GroupⅠwas taken as the control group, groupⅡas the hemorrhagic shock group, groupⅢas hemorrhagic shock laparotomy, groupⅣas hemorrhagic shock splenectomy and groupⅤas splenec-tomy omentectomy hemorrhagic shock group. Hemorrhagic shock was induced by drawing blood and reducing mean arterial pressure (MAP) to 40 mmHg within 10 min. After a hypotensive period of 1 h, animals were resuscitated. Bronchoalveolar lavage (BAL) was performed to recover cells from the alveolar space with 40 ml of BAL fluid after resuscitation malondialdehyde (MDA) and L-γ-glutarnyl-L-cysteinyl-glycine (GSH) levels were measured in serum, eryth-rocytes and lung tissue. RESULTS: Serum, erythrocyte, lung tissue MDA and GSH levels were significantly increased in hemorrhagic shock groupsⅡ-Ⅴ(P < 0.05). Lymphocyte, neutrophil and alveolar macrophage counts in BAL fluid indicated a significant difference between control and shock groups (P < 0.05). CONCLUSION: The degree of trauma increases hemorrhagic shock-induced acute lung injury.     

Expression and significance of MMP-9 and MDM2 in the oncogenesis of lung cancer in rats

Objective:To observe the expression of matrix metalloproteinase-9(MMP-9)and mouse double minute 2 homolog(MDM2)in the oncogenesis of lung cancer in rats and to explore their clinical value.Methods:A total of 140 rats were selected,of which 20 were selected randomly as the control group;and the remaining 120 as the observation group.The observation group was injected with benzopyrene to establish diseases model such as tissue proliferation,abnormal proliferation and lung cancer.Delected the MMP-9 levels of lung tissue by enzyme-linked assay,detected the MDM2 levels of lung tissue by immunochemistry assay.Results:The MMP-9 and MDM2 expression of the lung cancer group and the abnormal proliferation group were significantly higher than that in the tissue proliferation group and the control group,the difference was significant(P0.05).And the MDM2 expression of the tissue proliferation group was significantly higher than that in the control group,the difference was significant(P0.05).There was no significant difference in the MMP-9 expression between the tissue proliferation group and the control group(P0.05).The MDM2 and MMP-9 expression were increased in turn in the small cell carcinoma,squamous cell carcinoma and adenocarcinoma,the difference was statistically significant(P0.05).The MMP-9 and MDM2 expressions of stageⅢand stageⅣlung cancer tissue in rats were significant higher than that during stageⅠand stageⅡ,the difference was significant(P0.05).There was no significantly different in the MMP-9 and MDM2 expressions between stageⅢand stageⅣ(P0.05),and there is no significant difference of the MMP-9and MDM2 expressions between stageⅠand stageⅡ(P0.05).Conclusions:The expression of MMP-9 and MDM2 in lung tissue was associated with lung disease and lung cancer,both of them may be involved in the development and metastasis of lung cancer.Combined detection can be used as therapy and prognostic indicators for lung cancer.     

Protective effect of sodium tanshinon Ⅱ A on lung injury induced by limb ischemia-reperfusion in rats

Objective To investigate the protective effect of sodium tanshinone Ⅱ A on lung injury induced by limb ischemia-reperfusion in rats. Methods 60 SD rats were randomly divided into three groups:control group (group A,n =20),limb-ischemia-reperfusion group (group B,n =20),and tanshinone Ⅱ A group (group C,n =20). The levels of malondialdehyde (MDA),superoxide dismutase (SOD),xanthine oxidase (XOD),interleukin-8 (IL-8),and tumor necrosis factor-α (TNF-α) in plasma and lung tissue were measured. The levels of TXB2 and 6-kcto-PGF1α in serum were measured by radioimmunoassay. The morphological and ultrastructure changes of lung tissues were observed under light microscope and electron microscope,respectively. Results ①Under light microscope,compared with group B,expansion of pulmonary vascular,aggregation of inflammatory cell,pulmonary edema relieved in group C. Under electron microscopy,compared with group B,mitochondrial swelling or degeneration,edema of basement membrane,swelling of pulmonary capillary endothelial cell,and pulmonary interstitial edema relieved in group C. ②MDA and XOD in plasma and lung tissue of group B were significantly higher than those in group A (all P ＜0. 01),and those in group C were significantly lower than those in group B (all P ＜0. 01). SOD in plasma and lung tissue of group B was significantly lower than that in group A (all P ＜0. 01). and that in group C was significantly higher than that in group B (all P ＜0. 01). ③IL-8、TNF-α in serum and lung tissue of group B were significantly higher than those in group A (all P ＜0. 01),those in group C were lower than those in group B ( P ＜0. 05 or P＜0. 01),and there was no statistical significance between group C and group A. ④TXB2 and 6-keto-PGF1α in serum of group B were significantly higher than those in group A (all P ＜0. 01),and those in group C were significantly lower than those in group B (all P ＜0. 01). TXB2 in serum of group C was higher than that in group A ( P ＜0. 05). There was no statistical significance on serum 6-keto-PGF1α between group C and group A. Conclusions Tanshinone Ⅱ A can alleviate lung injury of rats with limb ischemia-reperfusion.     

Combined early fluid resuscitation and hydrogen inhalation attenuates lung and intestine injury

AIM:To study the effects of combined early fluid resuscitation and hydrogen inhalation on septic shockinduced lung and intestine injuries.METHODS:Wistar male rats were randomly divided into four groups:control group(Group A,n = 15);septic shock group(Group B,n = 15);early fluid resuscitation-treated septic shock group(Group C,n = 15);and early fluid resuscitation and inhalation of 2% hydrogentreated septic shock group(Group D,n = 15).The activity of hydroxyl radicals,myeloperoxidase(MPO),superoxide dismutase(SOD),diamine oxidase(DAO),and the concentration of malonaldehyde(MDA) in the lung and intestinal tissue were assessed according to the corresponding kits.Hematoxylin and eosin staining was carried out to detect the pathology of the lung and intestine.The expression levels of interleukin(IL)-6,IL-8,and tumor necrosis factor(TNF)-α in lung and intestine tissue were detected by enzyme-linked immunosorbent assay method.The expression levels of Fas and Bcl2 in lung tissues were determined by immunohistochemistry and Western blotting.RESULTS:Septic shock elicited a significant increase in the levels of MDA(10.17 ± 1.12 nmol/mg protein vs 2.98 ± 0.64 nmol/mg protein) and MPO(6.79 ± 1.02 U/g wet tissue vs 1.69 ± 0.14 U/g wet tissue) in lung tissues.These effects were not significantly decreased by Group C pretreatment,but were significantly reduced by Group D pretreatment(MDA:4.45 ± 1.13 nmol/mg protein vs 9.56 ± 1.37 nmol/mg protein;MPO:2.58 ± 0.21 U/g wet tissue vs 6.02 ± 1.16 U/g wet tissue).The activity of SOD(250.32 ± 8.56 U/mg protein vs 365.78 ± 10.26 U/mg protein) in lung tissues was decreased after septic shock,and was not significantly increased by Group C pretreatment,but was significantly enhanced by Group D pretreatment(331.15 ± 9.64 U/mg protein vs 262.98 ± 5.47 U/mg protein).Histological evidence of lung hemorrhage,neutrophil infiltration and overexpression of IL-6,IL-8,and TNF-α was observed in lung tissues,all of which were attenuated by Group C and further alleviated by Group D pretre     

Role of Kupffer cells in acute hemorrhagic necrotizing pancreatitis-associated lung injury of rats

AIM:To investigate the role of Kupffer cells(KCs)inacute hemorrhagic necrotizing pancreatitis-associatedlung injury(AHNP-LI).METHODS:Forty-two rats were allocated to fourgroups[sham operation,AHNP model,gadoliniumchloride(GdCl_3)pretreatment,GdCl_3 control].In GdCl_3pretreatment group,GdCl_3 was administered by caudalvein injection 24 h before the AHNP model induction.Blood from the iliac artery,alveolar macrophages andtissues from the pancreas and lung,were collected insix animals per group 3 and 6 h after acute pancreatitisinduction.TNF-α,IL-1 of serum,myeloperoxidase(MPO)of lung tissue,NF-kB activation of alveolar macrophageswere detected.Serum AST and ALT in sham operationgroup and GdCl_3 control group were tested.In addition,histopathological changes of the pancreas and lung wereobserved under light microscope.RESULTS:MPO of lung tissue and TNF-α,IL-1 levelsof serum were all reduced significantly in GdCl_3pretreatment group compared to those in AHNP group(P<0.01).NF-kB activation of alveolar macrophageswas also attenuated significantly in GdCl_3 pretreatmentgroup compared to that in AHNP group(P<0.01).Thepathological injury of the lung was ameliorated obviouslyin GdCl_3 pretreatment group compared to that in AHNPgroup.Nevertheless,the serum amylase level did notreduce and injury of the pancreas was not prevented inGdCl_3 pretreatment group.CONCLUSION:Pulmonary injury induced by AHNPis mediated by KC activation and AHNP-LI can be significantly ameliorated by pretreatment with GdCl_3 andKCs play a vital role in AHNP-LI.     

Effect of matrine on transforming growth factor β1 and hepatocyte growth factor in rat liver fibrosis model

Objective:To observe the preventive and control effect of matrine on transforming growth factor(TCF- β1) and hepatocyte.growth factor(HCF) of liver fibrosis tissue in rals.Methods:A total of48 SD rats were randomly divided into A,B,C,D groups with 12 in each,group A as the normal control group and groups B.C,D as liver fibrosis models using composite modulus method with carbon tetrachloride(CCL_4).Group B was the model group,group C adopted γ— interferon lavage therapy in the second day of modeling,and group D adopted matrine lavage treatment,at 4 and8 weeks after treatment.Six rats were executed for detection of TGF- β1 and HGF,liver tissue histology and comparison fibrosis degree changes of rat liver tissue between groups.Results:Croups B,C,D showed a more significantly increased TCF- β1 at each time point compared with group A(P0.05);Group B showed a more significantly increased TGF- β1 than groups C and D at weeks 4 and 8(P0.05);group D showed a lowest level of TGF-β1,followed by groups C and B.HGF of group B decreased more significantly than A group at weeks 4 and 8(P0.05);HGF of groups C and D was significantly elevated at 4 and 8 weeks than groups A and B(P0.05),in which the group D showed the highest level of HGF.According to tissue histologic observation,rat liver tissue structure of group A was clear and normal,tissue structure of group B was destroyed with obvious fibrous tissue hyperplasia and fatty change of hepatic cells;groups C and D showed a slighter liver tissue damage,cell necrosis and connective tissue hyperplasia in collect abbacy than group B with a trend of obvious improvement.Conclusions:Matrine can reduce TGF- β1expression and enhance the activity of HGF,so as to realize the inhibition effect on liver fibrosis in rats.     

Anti-tumor activity of tanshinone ⅡA in combined with cyclophosphamide against Lewis mice with lung cancer

Objective:To explore the anti-tumor activity of tanshinone ⅡA in combined with cyclophosphamide against Lewis mice with lung cancer and the effect on cellular immune function.Methods:Lewis tumor cells were inoculated suhcutaneously into the right armpit of mice in each group(n=20) to establish Lewis lung cancer mice model.After model establishment,mice in the model group were given normal saline by lavage,qd.Mice in treatment Ⅰ group were given intraperitoneal injection of TanIIA,15 mg/kg,qd.Mice in treatment Ⅱ group were given intraperitoneal injection of CTX,25 mg/kg,qd.Mice in treatment Ⅲ group were given intraperitoneal injections of TanIIA and CTX,in which the administration method of TanIIA was the same as in treatment Ⅰ group,continuously for 2 weeks,and the dosage of CTX was the same as in treatment Ⅱ group,24 h after model establishment,every other day.Mice were sacrificed 2 weeks after establishment.The tumor tissues were collected to calculate the anti-tumor rate.Immunohistochemistry was used to detect the expressions of Bcl-2,Bax,VEGF,Angiostatin,and Endostatin.FCM was used to detect T lymphocyte subsets in spleen and liver of mice.Results:The tumor weight in treatment Ⅰ,Ⅱ,and Ⅲ groups was significantly lower than that in the model group(P0.05).The tumor weight in treatment Ⅲ group was significantly lower than that in treatment Ⅰ and Ⅱ groups(P0.05).The anti-tumor rate in treatment Ⅱ and Ⅲ groups was significantly higher than that in treatment Ⅰ group(P0.05).Bcl-2 expression in the tumor tissues of treatment Ⅰ,Ⅱ,and Ⅲgroups was significantly lower than that in the model group(P0.05),while Bax expression was significantly higher than that in the model group(P0.05).Bcl-2 expression in the tumor tissues of treatment Ⅰ and Ⅱ groups was significantly higher than that in treatment Ⅲ group(P0.05),while Bax expression was significantly lower than that in treatment Ⅲ group(P0.05).CD4~+ and CD4~+/CD8~+ in treatment Ⅰ,Ⅱ,and Ⅲ groups were significantly higher than those in the model group(P0.05).CD4~+ in treatment Ⅲ group was significantly higher than that in treatment Ⅰ and Ⅱ groups(P0.05),while CD4~+/CD8~+ was significantly higher than that in treatment Ⅱ group(P0.05).The comparison of CD8~+ among each group was not statistically significant(P0.05).NK cell activity in treatment Ⅰ,Ⅱ,and Ⅲ groups was significantly higher than that in the model group(P0.05).NK cell activity in treatment Ⅲ group was significantly higher than that in treatment Ⅰ and Ⅱ groups(P0.05).Conclusions:TannA in combined with CTX can down regulate Bcl-2 expression in lung cancer tissues,up regulate Bax expression,inhibit the neovascularization of tumor tissues,and enhance the immunological function,with a significant anti-tumor activity.     

The mRNA expression patterns of tumor necrosis factor-α and TNFR-I in some vital organs after thermal injury

AIM: To investigate changes of tumor necrosis factor-α (TNFα) and TNFR-Ⅰ expression in vital organs and their significance in the pathogenesis of multiple organ damage associated with endogenous endotoxin following major burns.METHODS: Wistar rats subjected to a 35 % full-thickness scald injury were sacrificed at 12 h, 24 h, 48 h, and 72 h postburn, respectively. Meanwhile, eight rats were taken as normal controls. Tissue samples from liver, spleen, kidney,lung and intestine were collected to assay tissue endotoxin levels and measure TNF-α and TNFR-Ⅰ expression, In addition, blood samples were obtained for the determination of organ function parameters.RESULTS: Endotoxin levels in liver, spleen and lung increased markedly after thermal injury, with the highest level in liver. The gene expression of TNF-α in liver, lung and kidney was up-regulated after thermal injury, while the TNFR-Ⅰ mRNA expression in liver, lung, kidney and intestine was shown decreased throughout the observation period. Thus, the mRNA expression ratio of TNF-α to TNFRⅠ was significantly increased postburn, particularly in pulmonary tissue (67-fold). In addition, the significant correlations between the expression of TNFR-Ⅰ or the expression ratio of TNF-α/TNFR mRNA in liver tissue and serum aspartate aminotransferase levels were noted (P ＜0.05-0.01). Similar results were also obtained between pulmonary TNF-α mRNA expression and myeloperoxidase activities (P＜0.01), whereas there was a highly negative correlation between levels of renal TNFR-Ⅰ mRNA expression and serum creatinine.CONCLUSION: Burn injury could result in the translocation of gut-derived endotoxin that was mainly distributed in the liver, spleen and lung. The translocated endotoxin then made the expression of TNF-α and TNFR-Ⅰ mRNA up-regulated and down-regulated respectively in various organs, which might be involved in the pathogenesis of multiple organ damage following burns.     

Experimental study on the role of endotoxin in the development of hepatopulmonary syndrome

AIM: To evaluate the role of intestinal endotoxemia in the genesis of hepatopulmonary syndrome. METHODS: A rat model of cirrhosis was prepared with the method of compound factors. At the end of the eighth week, rats with cirrhosis were treated with 300 μg LPS/100 g body weight, and 1 g/rat of glycine about four h prior to LPS. After three h of LPS treatment, blood and tissues were collected for various measurements. Kupffer cells were isolated from male Wistar rats and cultured, and divided into five groups. Supernatant was harvested at 3 h after treatment with LPS for measurement of tumor necrosis factor-alpha (TNF-α). RESULTS: Our results showed that in rats with cirrhosis, slowed and deepened breath with occasional pause was. PaO2, PaCO2 and standard bicarbonate (SB) in arterial blood were decreased. Arterial O2 and actual bicarbonate (AB) were markedly decreased. There was a close correlation between decreased O2 and endotoxin. Metabolic acidosis accompanying respiratory alkalosis was the primary type of acid-base imbalance. The alveolar-arterial oxygen gradient was sharply widened. Massive accumulation of giant macrophages in the alveolar spaces and its wall and widened alveolar wall architecture were observed. The number of bacterial translocations in mesenteric lymph nodes increased. The ratio of TC99M-MAA brain-over-lung radioactivity rose. Endotoxin, and TNF-α, endothelin-1 (ET-1), nitric oxide (NO) in plasma and ET-1, carbon monoxide (CO) in lung homogenates increased. After administration of a given dosage of LPS in rats with cirrhosis, various pathological parameters worsened. Plasma level of endotoxin was related to TNF-α, ET-1, NO in plasma and ET-1, NO, CO in lung homogenates. TNF-α level was related to ET-1 and NO in plasma and lung homogenates and CO in lung homogenate as well. The level of TNF-α increased after infusion of LPS into culture supernatant of Kupffer cells in vitro. However, TNF-α significantly decreased after pretreatment with glycine, PD98059 and SB212850. Glycine could antagonize the effect of LPS in vivo and in vitro. CONCLUSION: Intestinal endotoxemia accompanying by cirrhosis may be an important mechanism in the development of hepatopulmonary syndrome in rats. Overproduction of TNF-α due to endotoxin stimulation of Kupffer cells via mitogen-activated protein kinase (MAPK) signal transduction pathway may be a major mechanism mediating the pathologic alterations of hepatopulmonary syndrome.     

Effect of alprostadil combined with Diammonium glycyrrhizinate on renal interstitial fibrosis in SD rats

Objective:To observe effect of alprostadil combined with Diammonium glycyrrhizinate on renal interstitial fibrosis in SD rate.Methods:A total of 75 SD rate were randomly divided into A,B,C,D,E groups with 15 in each group.Rats in group A served as the control group received just only but tissue separation without modeling operation,while model of unilateral ureteral obstruction(UUO) was established in B,C,D,E groups.Rats in A,B group were given saline lavage placebo treatment,while rats in C,D,E groups were given dianunonium glycyrrhizinate and alprostadil injection.Five rats were sacrificed 1,2,3 weeks after modeling,serum creatinine level of femoral venous blood was determined.Transforming growth factor- β1(TCF- β1) and concentration of connective tissue growth factor(CTGF) were also detected by using ELISA.Line renal interstitial tissue was taken after HE staining,renal interstitial TGF- β1 and CTGF expression were detected by using immunohistochemical method.Results:Serum creatinine levels of B,C,D,E group at different time points in were significantly higher than that of group A(P0.05);serum creatinine levels in group B were significantly higher than that of C,D,E group at each time point(P0.05).Serum creatinine level of Croup E was significantly lower than C,D group after 2,3 weeks(P0.05).Rate in A group at each time point showed no significant changes in TGF- β1 and CREA concentration in serum and kidney tissues(P0.05);while serum and kidney tissue TGF- β1,concentration of CREA.expression of rats in B,C,D,E groups showed a gradual increasing trend over time.TCF- β1 and CREF of Group B in serum and kidney tissues at each time point were significantly higher than that of the other groups(P0.05).TCF- β1 and CREF of Group E in serum and kidney tissues at each time point were significantly lower than that of B,C,D group at all time points in serum and kidney tissues(P0.05).Conclusions:Alprostadil combined with diammonium glycyrrhizinate can significantly lower the expression of TGF- β1 and CTGF in serum and tissues of SD rat with renal interstitial fibrosis,thus inhibit rat renal interstitial fibrosis process.It has synergy protective effect.     

Reproductive endocrinology in Hatano high- and low-avoidance rats during the estrous cycle

The high- and low-avoidance animals (HAA and LAA rats) were originally selected from Sprague-Dawley rats for their shuttle-box task. Reproductive endocrinology during the estrous cycle was compared between HAA and LAA rats. All HAA rats showed a regular 4-d estrous cycle, whereas most LAA rats (70.8%) showed a regular 5-d estrous cycle. The peak level of preovulatory luteinizing hormone (LH) surge level was significantly lower in LAA rats than in HAA rats on the day of proestrus. In contrast, the peak level of prolactin surge on the day of proestrus was significantly higher in LAA rats than in HAA rats. Plasma concentrations of follicle-stimulating hormone (FSH) and estradiol-17β were significantly lower in LAA rats as compared with HAA rats at 12 h on the day of estrus and from 24 h on the day of diestrus to 18 h on the day of proestrus. On the other hand, plasma concentrations of progesterone were significantly higher in LAA rats compared with HAA rats on the day of diestrus. The number of antral follicles (300–600 μm in diameter) at 12 h on the day of proestrus was significantly fewer in LAA rats than in HAA rats. The size and number of corpus luteum at 12 h on the day of estrus were significantly greater in LAA rats than in HAA rats. These results clearly demonstrated that apparent differences are observed in reproductive endocrinology between two Hatano strains. These strain differences probably originated from neural regulation of pituitary hormones.     

Effect of levocarnitine on TIMP-1,ICAM-1 expression of rats with coronary heart disease and its myocardial protection effect

Objective:To study the effect of levocarnitine(L-CN) on tissue inhibitor of metalloproteinase-1(TIMP-1) and intercellular adhesion molecule-1(ICAM-1) expression of rats with coronary heart disease and evaluate the protective effect of L-CN on myocardial cells.Methods:Highfat diet feeding and intraperitoneal injection of pituitrin was performed on rats in model group and CHD Model of rats was built.Rats with successful model-building were selected and divided into L-CN group and Ctrl group randomly.Rats in L-CN group were given L-CN treatment,with intraperitoneal injection of 200 mg·kg~(-1)?d~(-1) and successive administration for 3 d.Rats in Ctrl group were given equal volumes of normal saline.Blood was collected from carotid artery at different time and expression quantity of creatine kinase-MB(CK-MB) and Troponin Ⅰ(Tn Ⅰ)in serum was detected.Rats in each group were put to death and were separated to obtain the myocardial tissue.Real-time PCR and Western Blotting hybridization were performed to detect the TIMP-1.ICAM-1 expression in myocardial tissue in each group.Statistical analysis was employed to explore the expression changes of TIMP-1 and ICAM-1.and ELISA test was used lo analyse the expression changes of myocardial necrosis marker- CK-MB and Tn I to learn the effect of L-CN and its myocardial protective effect.Results:The total cholesterol,triglyceride and blood glucose levels of rats in model group were significantly higher than that in control group,which indicated that due to high-fat diet feeding,blood lipid of rats in model group was obviously higher than that in control group.In myocardial tissue of rats in model group,TIMP-I level significantly reduced and ICAM-1 level significantly increased(P0.0l).In model group,after L-CN treatment.TTMP-I level had double increase,while ICAM-1 level had 43%of decrease in L-CN group compared with Ctrl group.After L-CN intervention treatment.CK-MB and Tn Ⅰ content in L-CN group relatively reduced compared with Ctrl group.The difference among groups was obvious(P0.01).Condusions:L-CN could increase the TTMP-I expression level and inhibit the ICAM-1 expression level.L-CN has a certain myocardial protective effect     

Expression and significance of ICAM-1 and its counter receptors LFA-1 and Mac-1 in experimental acute pancreatitis of rats

AIM: To investigate the role of intercellular adhesion molecule-1 (ICAM-1) and its counter receptors LFA-1 and Mac-1 in acute pancreatitis (AP). METHODS: SD rats were allocated to AP group and control group randomly (25 rats each). AP was induced by infusion of 5% chenodeoxycholic acid into the pancreatic duct, followed by ligation of pancreatic duct. The rats were sacrificed at 1, 3, 6, 12 and 24 h after induction of pancreatitis. Five rats were sacrificed at one time point in the two groups before the blood and specimens from pancreas and lung were obtained. Serum amylase and ascitic fluid were measured at each time point. Expression of ICAM-1 at different time points was assessed by immunohistochemistry in pancreas and lung, and the expression of LAF-1 and Mac-1 on neutrophils at different time points was detected by flow cytometer. RESULTS: Induction of AP was confirmed by the serum levels of amylase and histological studies. The expression of ICAM-1 in pancreas increased significantly than that in the control group at all time points (P < 0.05 or P < 0.01), as well as the expression in lung except at 1 h. The expression of LFA-1 and Mac-1 on neutrophil in blood increased significantly in AP group than that in control group at several time points (P < 0.05 ox P < 0.01). The amount of ascitic fluid and serum amylase level of AP group increased significantly than that of control group at all time points (P < 0.05 or P < 0.01). Parallel to these results, a significant neutrophil infiltration was found in pancreas and lung tissues of AP group rats. CONCLUSION: Our findings suggest the important role for ICAM-1, LFA-1 and Mac-1 in mediating the development of AP from a local disease to a systemic illness. Upregulation of ICAM-1, LFA-1, Mac-1 and subsequent leukocyte infiltration appear to be significant events of pancreatic and pulmonary injuries in AP.     

Antioxidation of probucol in rats with experimental atherosclerosis

Background Oxidative stress plays an important role in atherogenesis,which raises the possibility of using antioxidants to ameliorate atherosclerosis.In this research,we aim to determine the effects of probucol on atherosclerosis in rats.Methods Forty-five male adult Wistar rats were randomly and equally allocated to three groups,control group(Group N),model group(Group M) and probucol group(Group P).High-lipid diet and intraperitoneal injection of Vitamin D3 were given to establish rats atherosclerosis(AS) model.Group P was intragastrically administered Probucol after 8 weeks.At the end of 16 weeks,all the rats were weighted and sacrificed for detecting the levels of triglycerides(TG),total cholesterol(TC),low-density lipoprotein(LDL),high-density lipoprotein(HDL),oxidized low-density lipoprotein(OX-LDL) and malonaldehyde(MDA) in serum and the activity of serum superoxide dismutase(SOD).The histomorphological changes of the aorta were observed under light microscope.Results The body weight of rats in Group M and Group P were lighter than that in Group N(P < 0.01),and rats in Group P were heavier than those in Group M(P < 0.01).The contents of TC,TG and LDL-C in serum were obviously elevated in Group M and Group P compared with those in Group N(P < 0.01),and the content of HDL in Groups M and P was lower than that in Group N(P < 0.01).The contents of TC and LDL-C in serum were significantly lower in Group P than those in Group M(P < 0.01).However,the contents of TG and HDL in Groups M and P showed no statistically significant differences with each other(P > 0.05).The serum OX-LDL and MDA levels significantly increased,SOD activity decreased in Groups M and P compared with Group N(P < 0.01),while the levels of OX-LDL and MDA in Group P were lower than those in Group M(P < 0.05).No vessel lesion was found in Group N.The endothelial damage of the aorta was more significantly severe in Group M than in Group N,while the vessel lesion was less severe in Group P than in Group M.Conclusion Probucol can prevent atherogenesis and play a crucial role in inhibition of oxidative stress.     

Therapeutic effect of interleukin-10 on CCI_4-induced hepatic fibrosis in rats

AIM: To study the therapeutic effect of exogenous inter-leukin-10 on CCI4-induced hepatic fibrosis in rats and its possible mechanisms. METHODS: Fourty-seven SD rats were randomly divided into control group (group N) and CCI4-induced hepatic fibrosis model group (group C). After CCI4 was given for 9 wk, the model group was divided into three groups. Rats in group M were put to death immediately, rats in group T were treated with IL-10 for another three wk and then put to death, rats in group R recovered after three weeks and were then killed. The degree of hepatic fibrosis was measured by HE staining and histological activity index (HAI). Histological activity index (HAI), change of collagen typesⅠandⅢwere measured by Picrosirius staining. The expression of TNF-α, MMP-2 and TIMP-1 in liver tissue was measured by S-P immunohis-tochemistry. RESULTS: CCI4- induced experimental rat hepatic fibrosis model was established successfully. The degree of hepatic fibrosis was markedly lower in group T than in groups M and R, and there was no difference between the two groups. The expression of collagen typesⅠandⅢwas significantly suppressed in group T and was slightly suppressed in groups M and R. The positive levels of TNF-α, MMP-2 and TIMP-1 in group M increased significantly compared to those in group N (P<0.01). The positive signals decreased significantly in groups T and R (P<0.01), but positive score was significantly lower in group T than in group R (P< 0.01). CONCLUSION: Exogenous IL-10 can reverse CCI4-in-duced hepatic fibrosis in rats. IL-10 may exert its reversible effects on hepatic fibrosis by blocking CCI4-induced inflammation, inhibiting expression of MMP-2 and TIMP-1 and promoting resolution of collagen typesⅠandⅢ.     

Defensive nature of Sargassum polycystum (Brown alga) against acetaminophen-induced toxic hepatitis in rats: Role of drug metabolizing microsomal enzyme system, tumor necrosis factor-a and fate of liver cell structural integrity

AIM: To assess the defensive nature of Sargassum polycystum (S. polycystum) (Brown alga) against acetaminophen (AAP)-induced changes in drug metabolizing mi crosomal enzyme system, tumor necrosis factor (TNF-α) and fine structural features of the liver during toxic hepatitis in rats. METHODS: Male albino Wistar strain rats used for the study were randomly categorized into 4 groups. GroupⅠconsisted of normal control rats fed with standard diet. GroupⅡrats were administered with acetaminophen (800 mg/kg body weight, intraperitoneally). GroupⅢrats were pre-treated with 5. polycystum extract alone. GroupⅣrats were orally pre-treated with S. polycystum extract (200 mg/kg body weight for 21 d) prior to acetaminophen induction (800 mg/kg body weight, intraperitoneally). Serum separated and liver was excised and microsomal fraction was isolated for assaying cytochrome P450, NADPH Cyt P450 reductase and bs. Serum TNF-αwas detected using ELISA. Fine structural features of liver were examined by transmission electron microscopy. RESULTS: Rats intoxicated with acetaminophen showed considerable impairment in the activities of drug metabolizing microsomal enzymes, such as cytochrome P450, NADPH Cyt P450 reductase and bs when compared with the control rats. The rats intoxicated with acetaminophen also significantly triggered serum TNF-αwhen compared with the control rats. These severe alterations in the drug metabolizing enzymes were appreciably prevented in the rats pretreated with 5. polycystum. The rats pretreated with S. polycystum showed considerable inhibition in the elevation of TNF-αcompared to the rats intoxicated with acetaminophen. The electron microscopic observation showed considerable loss of structural integrity of the endoplasmic reticulum, lipid infiltration and ballooning of mitochondria in the acetaminophen-intoxicated rats, whereas the rats treated with S. polycystum showed considerable protection against acetaminophen-induced alterations in structural integrity. CONCLUSION: These observations suggest that the animals treated with 5. polycystum extract may have the ability to protect the drug metabolizing enzyme system and mitochondrial functional status from free radical attack, thereby showing its defense mechanism in protecting hepatic cells from acetaminophen toxic metabolite N-acetyl-para-benzoquinone-imine (NAPQI).     

Hepatic fibrosis in biliary-obstructed rats is prevented by Ginkgo biloba treatment

AIM: To assess the antioxidant and antifibrotic effects of long-term Ginkgo biloba administration on liver fibrosis induced by biliary obstruction in rats. METHODS: Liver fibrosis was induced in male Wistar albino rats by bile duct ligation and scission (BDL). Ginkgo biloba extract (EGb 761, 50 mg/kg·per d) or saline was administered for 28 d. At the end of the treatment period, all rats were killed. Serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH) levels were determined to assess liver functions and tissue damage, respectively. Tumor necrosis factorα (TNF-α) was also assayed in serum samples. Liver tissues were taken for determination of the hepatic malondialdehyde (MDA) and glutathione (GSH) levels, myeloperoxidase (MPO) activity and collagen content. Production of reactive oxidants was monitored by chemiluminescence (CL) assay. Serum AST, ALT, LDH, and TNF-α levels were elevated in the BDL group as compared to control group and were significantly decreased by EGb treatment. RESULTS: Hepatic GSH level, depressed by BDL, was elevated back to control level in EGb-treated BDL group. Increase in tissue MDA level, MPO activity and collagen content due to BDL were also attenuated by EGb treatment. Furthermore, luminol and lucigenin CL values in BDL group increased dramatically compared to control and reduced by EGb treatment. CONCLUSION: Our results suggest that Ginkgo biloba protects the liver from oxidative damage following BDL in rats. This effect possibly involves the inhibition of neutrophil infiltration and lipid peroxidation; thus, restoration of oxidant and antioxidant status in the tissue.