IL-4 regulates specific Arg-1(+) macrophage sFlt-1-mediated inhibition of angiogenesis.

One of the main drivers for neovascularization in age-related macular degeneration is activation of innate immunity in the presence of macrophages. Here, we demonstrate that T helper cell type 2 cytokines and, in particular, IL-4 condition human and murine monocyte phenotype toward Arg-1(+), and their subsequent behavior limits angiogenesis by increasing soluble fms-like tyrosine kinase 1 (sFlt-1) gene expression. We document that T helper cell type 2 cytokine-conditioned murine macrophages neutralize vascular endothelial growth factor-mediated endothelial cell proliferation (human umbilical vein endothelial cell and choroidal vasculature) in a sFlt-1-dependent manner. We demonstrate that in vivo intravitreal administration of IL-4 attenuates laser-induced choroidal neovascularization (L-CNV) due to specific IL-4 conditioning of macrophages. IL-4 induces the expression of sFlt-1 by resident CD11b(+) retinal microglia and infiltrating myeloid cells but not from retinal pigment epithelium. IL-4-induced suppression of L-CNV is not prevented when sFlt-1 expression is attenuated in retinal pigment epithelium. IL-4-mediated suppression of L-CNV was abrogated in IL-4R-deficient mice and in bone marrow chimeras reconstituted with myeloid cells that had undergone lentiviral-mediated shRNA silencing of sFlt-1, demonstrating the critical role of this cell population. Together, these data establish how lL-4 directly drives macrophage sFlt-1 production expressing an Arg-1(+) phenotype and support the therapeutic potential of targeted IL-4 conditioning within the tissue to regulate disease conditions such as neovascular age-related macular degeneration.

[Prevention and treatment mechanism of qingxia therapy (based on yinchenhao decoction and dachengqi decoction) on hepatocyte apoptosis in rats with acute hepatic injury induced by lipopolysaccharide/D-galactosamine].

OBJECTIVE: To study the prevention and treatment mechanism of Qingxia therapy (based on Yinchenhao Decoction and Dachengqi Decoction) on hepatocyte apoptosis in rats with acute hepatic injury induced by lipopolysaccharide plus D-galactosamine (LPS/D-GalN). METHODS: The acute hepatic injury model was established by LPS/D-GalN and then intervened with Qingxia therapy. Serum liver function, PT and liver tissue pathology were observed, hepatocyte apoptosis index was detected by Tunel, protein expressions of BCL-2, BAX and Caspase-3 were detected by Western blotting. RESULTS: Qingxia therapy could significantly decrease serum ALT, AST and TBIL levels (P < 0.01 or 0.05), reduce hepatocyte necrosis and inflammatory cell infiltration. There were more apoptotic cells in model group, which had significant differences compared with Qingxia group and control group. Protein expressions of BAX and Caspase-3 in model group were significantly higher than those in control group and Qingxia group (P < 0.05), but BCL-2 protein expression in model group was lower (P < 0.05). CONCLUSION: Qingxia therapy can ameliorate the liver function and hepatic tissue pathology of rats with hepatic injury induced by LPS/D-GalN, alleviate hepatocyte apoptosis in rats, prevent and treat hepatocyte apoptosis by down-regulating the protein expressions of Caspase-3 and BAX, up-regulating the protein expression of BCL-2, and adjusting the balance of BCL-2/BAX.

Ginsenoside Rb1 reverses H2O2-induced senescence in human umbilical endothelial cells: involvement of eNOS pathway.

OBJECTIVE: Senescence of endothelial cells has been implicated in endothelial dysfunction and atherogenesis. This study investigated the effects of Rb1, a major ginsenoside in ginseng, on H2O2-induced senescence in primary human umbilical vein endothelial cells (HUVECs). METHODS AND RESULTS: Real-time PCR and Western blot were used to detect the mRNA and protein expression, respectively. H2O2 (40∼100 µmol/L) effectively increased SA-ß-gal activity and PAI-1 mRNA levels, two important senescence related biomarkers, in HUVECs, which were dramatically inhibited by Rb1 pre-incubation. Furthermore, Rb1 administration reversed the H2O2-decreased protein and mRNA levels of eNOS and its phosphorylation at Ser-1177, and the increased eNOS phosphorylation at Thr-495. As a result, Rb1 pretreatment restored the NO generation, as assayed by nitrate reductase method. However, pretreatment with L-NAME, a NOS inhibitor, abolished all the inhibitory effects of Rb1 on senescence. Importantly, Rb1 modulated the H2O2-altered caveolin-1 and pAkt, two most important regulators of eNOS expression and activity, in HUVECs. CONCLUSIONS: We showed that Rb1 effectively protects HUVECs from senescence through eNOS modulation.

Potential advantages of a combination of chinese medicine and bone marrow mesenchymal stem cell transplantation for removing blood stasis and stimulating neogenesis during ischemic stroke treatment.

Combined treatment of ischemic stroke with Chinese medicine and exogenous bone marrow mesenchymal stem cell (BMSC) transplantation may improve the removal of blood stasis and stimulation of neogenesis. Chinese medicines that remove blood stasis not only promote blood circulation but also calm the endopathic wind, remove heat, resolve phlegm, remove toxic substances and strengthen body resistance. The medicinal targeting effect of Chinese medicine can promote the homing of BMSCs, and the synergistic therapeutic effects of drugs can contribute to BMSC differentiation. As such, exogenous BMSC transplantation has potential advantages for neogenesis. Chinese medicines and exogenous BMSCs provide complementary functions for the removal of blood stasis and tion of Chinese medicine and transplantation of exogenous BMSCs may be particularly suited to ischemic stroke treatment.

Rb1 protects endothelial cells from hydrogen peroxide-induced cell senescence by modulating redox status.

Senescence of endothelial cells has been proposed to play an important role in endothelial dysfunction and atherogenesis. In the present study we aimed to investigate whether ginsenoside Rb1, a major constituent of ginseng, protects endothelial cells from H(2)O(2)-induced endothelial senescence. While H(2)O(2) induced premature senescent-like phenotype of human umbilical vein endothelial cells (HUVECs), as judged by increased senescence-associated ß-galactosidase (SA-ß-gal) activity, enlarged, flattened cell morphology and sustained growth arrest, our results demonstrated that Rb1 protected endothelial cells from oxidative stress induced senescence. Mechanistically, we found that Rb1 could markedly increase intracellular superoxide dismutase (Cu/Zn SOD/SOD1) activity and decrease the malondialdehyde (MDA) level in H(2)O(2)-treated HUVECs, and suppress the generation of intracellular reactive oxygen species (ROS). Consistent with these findings, Rb1 could effectively restore the protein expression of Cu/Zn SOD, which was down-regulated in H(2)O(2) treated cells. Taken together, our data demonstrate that Rb1 exhibits antioxidant effects and antagonizes H(2)O(2)-induced cellular senescence.

Ginsenoside-Rg1 mediates microenvironment-dependent endothelial differentiation of human mesenchymal stem cells in vitro.

Bone marrow-derived mesenchymal stem cells (MSCs) possess a multi-lineage differentiation potential and have the ability to repair and rebuild injured vessels. The autologous differentiated MSC transplantation also makes possible the tissue-engineered grafts. Therefore, the efficient endothelial differentiation of MSCs could be beneficial in the successful injured vessel repair and engraftment. Ginsenoside-Rg1, the most prevalent active constituent of ginseng, is a potent proangiogenic factor of vascular endothelial cells and also has the ability to enhance the proliferation of bone marrow cells. The aim of this study is to investigate the role of ginsenoside-Rg1 in the microenvironment-dependent endothelial differentiation of human MSCs (hMSCs) in vitro. The endothelial differentiation environment was established by co-culturing hMSCs with mature endothelial cells (human umbilical vein endothelial cells) indirectly in vitro. Reverse transcriptase-polymerase chain reaction analysis and fluorescence immunocytochemistry showed a strong expression of endothelial-specific markers such as CD31, Von Willebrand factor, and VE-cadherin. Electron microscopy showed the endothelial characteristic Weibel-Palade bodies of differentiated hMSCs. The increased expression of CD31 demonstrated that Rg1 promoted the endothelial differentiation of hMSCs. The findings here show the differentiation of hMSCs into cells with phenotypic features of endothelial cells using indirect co-culture with mature endothelial cells and provide the evidence that ginsenoside-Rg1 can promote the milieu-dependent endothelial differentiation of hMSCs in vitro.

[Identification of salivary biomarkers in breast cancer patients with thick white or thick yellow tongue fur using isobaric tags for relative and absolute quantitative proteomics].

OBJECTIVE: To explore the presence of informative protein biomarkers in the salivary proteome of breast cancer patients with thick white or thick yellow tongue fur. METHODS: Salivia samples were collected from 20 breast cancer patients with thick white or yellow tongue fur and 10 healthy controls. The samples were profiled by using isobaric tags for relative and absolute quantitation (iTRAQ) technology coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). The analyzed map and data were assessed with Mascott 2.2 and Scaffold software. Ratio of proteins between groups of less than 0.6 or more than 1.5 could confirm that there was difference between groups. RESULTS: A total of 464 proteins were identified and 125 proteins met strict quantitative criteria. There were 9 proteins associated with breast cancer, expression levels of which were up- or down-regulated more than 1.5 folds compared with healthy people. There were 16 proteins associated with tongue coating, of which 10 proteins expressed in breast cancer patients with thick white fur were lower than in patients with thick yellow fur, and the expressions of the other 6 proteins were increased. CONCLUSION: This study demonstrates that iTRAQ combined with LC-MS/MS quantitative proteomics is a powerful tool for biomarker discovery and the identification of proteins associated with breast cancer and tongue coating.

Alterations in the Composition of Intestinal DNA Virome in Patients With COVID-19.

Patients with Coronavirus Disease 2019 (COVID-19), due to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection mainly present with respiratory issues and related symptoms, in addition to significantly affected digestive system, especially the intestinal tract. While several studies have shown changes in the intestinal flora of patients with COVID-19, not much information is available on the gut virome of such patients. In this study, we used the viromescan software on the latest gut virome database to analyze the intestinal DNA virome composition of 15 patients with COVID-19 and investigated the characteristic alternations, particularly of the intestinal DNA virome to further explore the influence of COVID-19 on the human gut. The DNA viruses in the gut of patients with COVID-19 were mainly crAss-like phages (35.48%), Myoviridae (20.91%), and Siphoviridae (20.43%) family of viruses. Compared with healthy controls, the gut virome composition of patients with COVID-19 changed significantly, especially the crAss-like phages family, from the first time of hospital admission. A potential correlation is also indicated between the change in virome and bacteriome (like Tectiviridae and Bacteroidaceae). The abundance of the viral and bacterial population was also analyzed through continuous sample collection from the gut of patients hospitalized due to COVID-19. The gut virome is indeed affected by the SARS-CoV-2 infection, and along with gut bacteriome, it may play an important role in the disease progression of COVID-19. These conclusions would be helpful in understanding the gut-related response and contribute to the treatment and prevention strategies of COVID-19.

Fuzi polysaccharide-1 produces antidepressant-like effects in mice.

Current antidepressants are clinically effective only after several weeks of administration. We show that Fuzi polysaccharide-1 (FPS), a new water-soluble polysaccharide isolated from Fuzi, which has been used to treat mood disorders in traditional Chinese medicine for centuries, increases the number of newborn cells in the dentate gyrus in adult mice, and most of these cells subsequently differentiate into new neurons. We also found that FPS administration reduces immobility in the forced swim test, and latency in the novelty suppressed-feeding test. Moreover, a 14-d regimen with FPS reverses avoidance behaviour and inhibition of hippocampal neurogenesis induced by chronic defeat stress. In contrast, imipramine, a well known antidepressant, reverses this avoidance behaviour only after 4 wk of continuous administration. Finally, acute treatment with FPS had no effect on brain monoamine levels in frontal cortex but significantly increases BDNF in the hippocampus, while the antidepressant effect and enhancement of cell proliferation induced by FPS administration were totally blocked by K252a, an inhibitor of trkB in a chronic social defeat depression model, suggesting that the neurogenic and antidepressant effects of FPS may involve BDNF signalling. In conclusion, our findings suggest that FPS could be developed as a putative antidepressant with a rapid onset of action.

Homocysteine impaired endothelial function through compromised vascular endothelial growth factor/Akt/endothelial nitric oxide synthase signalling.

1. Hyperhomocysteinaemia (HHcy) is associated with endothelial dysfunction and has been recognized as a risk factor of cardiovascular disease. The present study aimed to investigate the effect of homocysteine (Hcy) on endothelial function in vivo and in vitro, and the underlying signalling pathways. 2. The HHcy animal model was established by intragastric administration with l-methionine in rats. Plasma Hcy and nitric oxide (NO) concentration were measured by fluorescence immunoassay or nitrate reductase method, respectively. Vasorelaxation in response to acetylcholine and sodium nitroprusside were carried out on aortic rings. Human umbilical vein endothelial cells (HUVEC) were treated with indicated concentrations of Hcy in the in vitro experiments. Intracellular NO level and NO concentration in culture medium were assayed. The alterations of possible signalling proteins were detected by western blot analysis. 3. l-methionine administration induced a significant increase in plasma Hcy and decrease in plasma NO. Endothelium-dependent relaxation of aortic rings in response to acetylcholine was impaired in l-methionine-administrated rats. The in vitro study showed that Hcy reduced both intracellular and culture medium NO levels. Furthermore, Hcy decreased phosphorylation of endothelial nitric oxide synthase (eNOS) at serine-1177 and phosphorylation of Akt at serine-473. Hcy-induced dephosphorylation of eNOS at Ser-1177 was partially reversed by insulin (Akt activator) and GF109203X (PKC inhibitor). Furthermore, Hcy reduced vascular endothelial growth factor (VEGF) expression in a dose-dependent manner. 4. In conclusion, Hcy impaired endothelial function through compromised VEGF/Akt/endothelial nitric oxide synthase signalling. These findings will be beneficial for further understanding the role of Hcy in cardiovascular disease.

Fasudil attenuates lipopolysaccharide-induced acute lung injury in mice through the Rho/Rho kinase pathway.

BACKGROUND: The transendothelial migration of polymorphonuclear leukocytes (PMNs, neutrophils) may be a hallmark of acute lung injury (ALI). The breakdown of the vascular endothelial barrier has likewise been considered to have etiologic linkage in the pathogenesis of ALI. Rho-associated coiled-coil-forming protein kinase (ROCK), a downstream target effector of the small GTP-binding protein Rho, plays a key role in cell adhesion, motility, and contraction mediated by reorganization of the actin cytoskeleton. The aims were to investigate protection by fasudil in lipopolysaccharide (LPS)-induced ALI and the role of ROCK2 in neutrophil transendothelial migration in a murine model. MATERIAL/METHODS: Mice were assigned to three groups: sham-treated controls (Sham group), LPS instillation (LPS group), and protective application of fasudil and LPS instillation (Fasudil/LPS group). Indexes tested were breathing frequency, histopathological examination, lung injury score, lung wet-to-dry weight ratio, neutrophil percentage in bronchoalveolar lavage fluid (BALF), myeloperoxidase activity, and ROCK2 mRNA expression in lung homogenate. RESULTS: Permeability pulmonary edema (histopathological examination, lung injury score, and lung wet-to-dry weight ratio) was ameliorated and neutrophil infiltration in the lungs (neutrophil percentage in BALF, myeloperoxidase activity) was depressed in response to fasudil administration. Expression of ROCK2 mRNA in the lung homogenates of the LPS-treated mice increased approximately fourfold; however, fasudil did not affect the increase. CONCLUSIONS: The Rho/Rho kinase pathway may play an important role in the pathogenesis of LPS-induced ALI and fasudil, as a ROCK inhibitor, could decrease neutrophil transendothelial migration by attenuating cytoskeletal rearrangement of endothelial cells, leading to the inhibition of ALI development.

[Protective effects of sini decoction on adriamycin-induced heart failure and its mechanism].

OBJECTIVE: To investigate the protective effects of Sini decoction (SND) on Adriamycin-induced heart failure and its mechanism. METHODS: SD rats were randomly divided into three groups:control group,heart failure model group and SND group. ADR was injected in to the rats of model group and SND group by caudal vein. After injection,the rats in SND group were given SND [3.75 g/(kg x d), p.o.]. Three weeks later, protein expressions of Bid and Bcl-xl were detected by immunohistochemistry; mRNA expression ratio of Bcl-xl/Bcl-xs was detected by RT-PCR and apoptosis rate was determinated by flow cytometry. RESULTS: Compared with control group, the protein expression of Bcl-xl and mRNA ratio of Bcl-xl/Bcl-xs obviously decreased,while the protein expression of Bid and apoptosis rate significantly increased in the model group. SND could decrease cell apoptosis, increase the protein expression of Bcl-xl, increase bcl-xl/bcl-xs mRNA ratio and decrease Bid protein expression. CONCLUSION: Bcl-xl may play an important role in ADR-induced heart failure rats. The mechanism of SND on protecting cardiocyte may be related to apoptosis correlation factor, Bcl-xl and Bid.

Treatment of Uncomplicated Recurrent Urinary Tract Infection with Chinese Medicine Formula: A Randomized Controlled Trial.

OBJECTIVE: To evaluate Chinese medicine (CM) formula Bazheng Powder () as an alternative therapeutic option for female patients with recurrent urinary tract infection (RUTI). METHODS: A randomized double-blinded trial was performed. Eligible female patients with RUTI were recruited from one hospital and two community health centers. By using a blocked randomization scheme, participants were randomized to receive a CM formula (10 herbs) for 4 weeks or antibiotics for 1 week, followed by 3 weeks of placebo. Clinical cure rate and microbiological cure and recurrence after treatment were evaluated. RESULTS: A total 122 eligible patients were enrolled, with 61 cases in each group. The clinical cure rate by the intentto- treatment approach was 90.2% for the CM group and 82.0% for the antibiotics group (P>0.05). Bacteria were cleared from 88.5% (54/61) of patients in the CM group and 82.0% (50/61) in the antibiotics group. The recurrence rate in recovered patients at the 6-month follow-up was 9.1% (5/61) and 14.0 (7/61) in the CM and antibiotics groups, respectively (P>0.05). CONCLUSION: CM formula Bazheng Powder is a good alternative option for RUTI treatment. (Registration No. NCT01745328).

Ginsenoside Rb1 preconditioning protects against myocardial infarction after regional ischemia and reperfusion by activation of phosphatidylinositol-3-kinase signal transduction.

BACKGROUND: Ginsenoside Rb1, a major bioactive component of Panax ginseng, bears various beneficial effects on the cardiovascular system. This study investigated whether ginsenoside Rb1 preconditioning has protective effects on myocardial ischemia-reperfusion injury and its potential mechanism. METHODS: Rats subjected to 45 min of myocardial ischemia followed by 120 min of reperfusion were assigned to the following groups: sham-operated, ischemia-reperfusion (I/R), ginsenoside Rb1+I/R, wortmannin(a specific PI3K inhibitor)+I/R, wortmannin drug vehicle (dimethyl sulfoxide, DMSO), wortmannin+sham, ginsenoside Rb1+ wortmannin +I/R. Infarct size was assessed by triphenyltetrazolium chloride staining. Plasma creatine kinase (CK), creatine kinase isoenzyme MB (CK-MB), lactate dehydrogenase (LDH), and troponin T levels were also measured. Akt phosphorylation expression was assessed by immunoblotting. RESULTS: Ginsenoside Rb1 preconditioning reduced infarct size compared with that in the I/R group: 30 +/- 2.6% versus 51 +/- 2.7% (p < 0.01). Ginsenoside Rb1 preconditioning also markedly reduced the plasma CK, CK-MB, LDH and troponin T levels in blood. Akt phosphorylation expression increased after ginsenoside Rb1 preconditioning. These effects of ginsenoside Rb1 preconditioning were significantly inhibited by wortmannin. CONCLUSION: This is the first study to demonstrate that ginsenoside Rb1 preconditioning has protective effects on myocardial ischemia and reperfusion injury, partly by mediating the activation of the PI3K pathway and phosphorylation of Akt.

[The effect of PKC activation and Smac release on inhibition of myocardial cell apoptosis by Sini Decoction].

OBJECTIVE: To investigate the effect of Sini Decoction on myocardial cell apoptosis after MI/R and the possible relation to myocardial cell apoptosis, PKC and Smac. METHODS: Sprage-Dawley rats were divided into four groups: control group, sham group, MI/R group and SND group. The rats in MI/R group, left anterior descending coronary artery (LAD) was occluded for 1 h and reperfused for 1 h. The rats in SND group were pretreated with Sini decoction(5 g/kg x d) for three days, the last treatment was pretreated 24 h before the index occlusion. Cell apoptosis was measured by flow cytometry, the expression of PKC was measured by immunohistochemistry. The expression of Smac was analyzed by Western blotting. The activity of Caspase-3 was also measured. RESULTS: 24 h after Sini Decoction treatment, Myocardial cell apoptosis significantly decreased, PKC was translocated and its expression increased, the release of Smac decreased, the activity of Caspase-3 decreased. CONCLUSION: Sini Decoction treatment can protect myocardial cell from apoptosis caused by MI/R, the mechanism is related to PKC activation, Mitochondria protection and in turn inhibition of Smac release, and inhibition of apoptosis signal transduction.

[Study on activity and mechanism of Sini Decoction anti-mitochondrial oxidation injury caused by myocardial ischemia/reperfusion].

OBJECTIVE: To study the activity and mechanism of Sini Decoction (SND) Anti-mitochondrial Oxidation Injury caused by Myocardial Ischemia/Reperfusion. METHODS: Kun ming mice were randomly divided into three groups: Control group, Ischemia/Reperfusion (I/R) group and SND-treated group. At the end of experiment,hearts of mice were taken out for further detection. Activity of myocardium and mitochondrial SOD, content of myocardium and mitochondrial MDA, swelling of mitochondria, Lactic Acid content of myocardium and MnSODmRNA expression were observed. RESULTS: SND could increase the activity of myocardium and mitochondrial SOD (P<0.01), decrease the content of myocardium and mitochondrial MDA (P<0.01), decrease the Lactic Acid content of myocardium, lighted the swelling of mitochondria (P<0.01) and change the expression of MnSODmRNA (P<0.01). CONCLUSION: Sini decoction can anti-mitochondrial oxidation injury caused by Myocardial Ischemia/Reperfusion, its mechanism may be relate to increasing the MnSODmRNA expression.

Traditional Chinese Medicine Syndrome Patterns and Their Association with Hepatitis B Surface Antigen Levels during the Natural History of Chronic Hepatitis B Virus Infection.

The aim of this study is to investigate traditional Chinese medicine syndrome (TCMS) patterns and their association with hepatitis B surface antigen (HBsAg) levels during the natural history of chronic hepatitis B virus infection (CHB). Patients were categorized according to the phase of CHB, as follows: immune tolerance (ITP); immune clearance (ICP); low or nonreplication (LRP); reactivation (RAP); hepatic cirrhosis (HC); and primary liver cancer (PLC). TCMS patterns were classified among the following types: spleen-kidney deficiency (SKD); liver-qi depression (LQD); damp-heat in liver-gallbladder (LGDH); liver-kidney deficiency (LKD); and blood stasis blocking collateral (BSBC). HBsAg levels and other serological indicators were quantified for all patients and their association with TCMS was statistically analyzed and determined. Two hundred and eighty-nine patients with CHB were included. During the natural history of CHB, TCMS patterns were statistically different among the different phases (P < 0.001). The most frequently occurring syndromes among the six progressive phases were SKD, LGDH, LKD, LGDH, BSBC, and LGDH, respectively. The predominant patterns in the inactive stage (ITP + LRP), active stage (ICP + RAP), and late or advanced stage (HC + PLC) were SKD (31%), LGDH (51.8%) and BSBC (34.4%), respectively. Median HBsAg levels were also statistically different among the five patterns of TCMS (P < 0.001). The highest HBsAg levels were observed in SKD (4.48 log10 IU/mL). Medium levels were in LQD (3.91 log10 IU/mL) and LGDH (3.90 log10 IU/mL). The lowest HBsAg levels were in LKD (3.60 log10 IU/mL) and the second lowest levels in BSBC (3.81 log10 IU/mL). In addition, HBsAg levels in LKD and BSBC were significantly lower than those in SKD, LQD, and LGDH (P < 0.05 or 0.001). TCMS was altered during the natural history of CHB and correlated with HBsAg titers. This study could provide further insight into the therapy of CHB.

Ginkgo biloba extract (EGb 761) attenuates lung injury induced by intestinal ischemia/reperfusion in rats: roles of oxidative stress and nitric oxide.

AIM: To investigate the effect of ginkgo biloba extract (EGb 761) on lung injury induced by intestinal ischemia/reperfusion (II/R). METHODS: The rat model of II/R injury was produced by clamping the superior mesenteric artery for 60 min followed by reperfusion for 180 min. The rats were randomly allocated into sham, II/R, and EGb + II/R groups. In EGb + II/R group, EGb 761 (100 mg/kg per day) was given via a gastric tube for 7 consecutive days prior to surgery. Rats in II/R and sham groups were treated with equal volumes of the vehicle of EGb 761. Lung injury was assessed by light microscopy, wet-to-dry lung weight ratio (W/D) and pulmonary permeability index (PPI). The levels of malondialdehyde (MDA) and nitrite/nitrate (NO2(-)/NO3(-)), as well as the activities of superoxide dismutase (SOD) and myeloperoxidase (MPO) were examined. Western blot was used to determine the expression of inducible nitric oxide synthase (iNOS). RESULTS: EGb 761 markedly improved mean arterial pressure and attenuated lung injury, manifested by the improvement of histological changes and significant decreases of pulmonary W/D and PPI (P < 0.05 or 0.01). Moreover, EGb 761 markedly increased SOD activity, reduced MDA levels and MPO activity, and suppressed NO generation accompanied by down-regulation of iNOS expression (P < 0.05 or 0.01). CONCLUSION: The results indicate that EGb 761 has a protective effect on lung injury induced by II/R, which may be related to its antioxidant property and suppressions of neutrophil accumulation and iNOS-induced NO generation. EGb 761 seems to be an effective therapeutic agent for critically ill patients with respiratory failure related to II/R.

The effect of ginkgo biloba extract (EGb 761) pretreatment on intestinal epithelial apoptosis induced by intestinal ischemia/reperfusion in rats: role of ceramide.

Apoptosis was demonstrated to be a major mode of intestinal epithelial cell death caused by intestinal ischemia/reperfusion (II/R). Ceramide has been proposed as a messenger for apoptosis. The present study was aimed to investigate the effect of Ginkgo biloba extract 761 (EGb 761) pretreatment on II/R-induced intestinal mucosal epithelial apoptosis in rats and the mechanism related to ceramide. The rat model of II/R injury was produced by clamping superior mesenteric artery for 60 min followed by reperfusion for 180 min. Twenty four rats were randomly allocated into Sham, II/R and EGb + II/R groups. In EGb + II/R group, EGb 761 (100 mg/kg per day) was administered intragastrically for 7 days before the surgery. Animals in II/R and sham groups were treated with equal volume of normal saline solution. Intestinal mucosal epithelial apoptosis was detected via electron microscopy and TUNEL method. Lipid peroxidation in intestinal mucosa was determined by detecting the malondialdehyde level and the activities of superoxide dismutase and peroxidase glutathione. The ceramide generation and sphingomyelinase (SMase) mRNA expression in intestinal mucosa were determined by high performance, thin layer chromatography, and RT-PCR, respectively. II/R caused intestinal mucosal epithelial apoptosis and over-production of the ceramide accompanied by up-regulation of SMase mRNA expression and increases of lipid peroxidation. EGb 761 pretreatment significantly decreased apoptosis index, and concurrently reduced the ceramide generation accompanied by down-regulation of SMase expression and inhibition of lipid peroxidation. The findings indicate that EGb 761 pretreatment attenuates II/R-induced intestinal epithelial apoptosis, which might be attributable to its antioxidant action of mediating ceramide pathway.