Effects of berberine and yohimbine on splenocyte apoptosis in septic mice

AIM: To observe the effects of berberine and yohimbine on splenocyte apoptosis in septic mice and underlying mechanisms. METHODS: The mice were subjected to cecal ligature and puncture (CLP). The drugs or vehicle were given intragastrically 2 h after the surgery according to the following 5 groups: sham, CLP, CLP+berberine, CLP+yohimbine, and CLP+berberine+yohimbine. The apoptosis of splenocytes stained by TUNEL was observed under laser scanning confocal microscope 20 h after CLP. The splenic lymphocytes were isolated and observed using flow cytometry. The activities of caspase-3, caspase-8 and caspase-9 in splenic lymphocytes were detected, and the expression of Fas, Bim, Bcl-2 and Bax in the splenocytes was also determined by Western blotting. RESULTS: The TUNEL staining showed that the apoptotic rate of the splenocytes in septic mice 20 h after CLP was significantly higher than that in sham and CLP+yohimbine groups (P＜0.05). Compared with CLP group, the proportion of apoptotic cells was decreased in septic mice in CLP+berberine+yohimbine and CLP+yohimbine groups (P＜0.05). Flow cytometry analysis demonstrated the similar results in the apoptosis of splenocytes and T lymphocytes. However, only yohimbine treatment reduced the apoptosis of B lymphocytes in the spleen of sepsis-challenged mice. Compared with CLP group, caspase-9 activity was significantly reduced in CLP+berberine group (P＜0.05), the activities of caspase-3, caspase-8 and caspase-9 were all statistically reduced (P＜0.05) in CLP+yohimbine group and CLP+yohimbine+berberine group. CLP significantly increased the expression of cytosolic Fas, Bim and mitochondrial Bax in the splenocytes, and decreased Bcl-2 expression compared with sham group. Compared with CLP group, the expression of cytosolic Bim and mitochondrial Bax in CLP+berberine group were reduced (P＜0.05). Fas expression decreased only in CLP+yohimbine group (P＜0.05). Berberine combined with yohimbine reduced the expression of cytosolic Fas, Bim and mitochondrial Bax in the septic mouse splenocytes (P＜0.05).CONCLUSION: Yohimbine reduces sepsis-induced splenic lymphocyte apoptosis in mice by inhibiting Fas expression and in turn blocking both extrinsic and intrinsic apoptosis pathways. Berberine reduces Bim expression and inhibits caspase-9 activation, but not caspase-3 activation and apoptosis in the septic mouse splenocytes. Berberine combined with yohimbine reduces splenocyte apoptosis in the septic mice by inhibiting both extrinsic and intrinsic apoptotic pathways.     

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

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

Expression of apoptosis signal protein induced by Fas in labial salivary gland of patients with primary Sjgren’s syndrome

AIM:To study the expression of apoptosis signal proteins induced by Fas in labial salivary gland of patients with primary Sjgren’s syndrome (pSS), and investigate the possible pathway of the cell signaling transduction in pSS. METHODS:Biopsies of minor submucosal labial salivary gland were obtained from 32 patients with pSS and 8 normal controls. Fas, FasL and FADD were detected by Western blotting. Caspase-3 was measured by immunohistochemistry and CAD mRNA expression was determined by RT-PCR. RESULTS:The expressions of Fas, FasL, FADD, caspase-3 and CAD mRNA in labial salivary glands of patients with pSS were significantly higher than those in control group (P＜0.05). CONCLUSION:The apoptosis signalling transduction in labial salivary glands of patients with pSS may be that the product of FasL combined with Fas activates caspase-8 through FADD, then ICE family is cascaded, and finally CAD is splited by caspase-3 from ICAD, which catalyzes the degradation of DNA.      

Effect of puerarin on expression of Fas/FasL mRNA in lung tissue with pulmonary injury induced by ischemia-reperfusion in rabbits

AIM：To investigate the effect of puerarin (Pur) on expression of Fas/FasL mRNA in lung tissue during pulmonary ischemia and reperfusion injury (PIRI) in rabbits.METHODS：Single lung ischemia and reperfusion animal model was used.The rabbits were randomly divided into three groups,sham operated group (sham,n=10),PIR group (I-R,n=30) and PIR+ Pur group (Pur,n=30).Changes of several parameters included apoptotic index (AI),wet to dry ratio of lung tissue weight (W/D) and index of quantitative assessment of histologic lung injury (IQA) were measured at 60,180 and 300 minutes after reperfusion in lung tissue.Meanwhile,the location and expression of Fas/FasL mRNA were observed.Lung tissue was prepared for light microscopic and electron microscopic observation at 60,180,300 minutes after reperfusion.RESULTS：As compared with group I-R,Fas/FasL mRNA slightly expressed in intima and extima of small pulmonary artery,alveoli,and bronchiole epithelia in group Pur.The values of AI,W/D and IQA showed significantly lower than that in group I-R at 60,180,300 minutes after reperfusion in lung tissue (P<0.01 and P<0.05).Meanwhile,abnormal changes of the lung tissue in morphologically were lessen markedly in group Pur.CONCLUSION：Puerarin produces a notable protective effects on PIRI in rabbits by inhibiting Fas/FasL mRNA expression and decreasing apoptosis.     

Effects of panax notoginseng saponins on pneumocyte apoptosis and Fas/FasL expression in rabbits with lung ischemia/reperfusion injury

AIM: AIM: To explore the relationship between apoptosis in the lung tissues and lung ischemia/reperfusion injury, and observe effects of panax notoginseng saponins (PNS) on apoptosis in lung ischemia/reperfusion injury. METHODS: Single lung in situ ischemia/reperfusion animal model was used. Eighty four Japanese white rabbits were randomly divided into control group (control), ischemia/reperfusion 1 h group (IR1h), IR3h, IR5h, Panax Notoginseng Saponins 1 h group (PNS1h), PNS3h and PNS5h. TUNEL, immunocytochemistry and in situ hybridization techniques were used to observe apoptosis and Fas/FasL expression in various phases of lung ischemia/reperfusion. RESULTS: Cell apoptosis in lung tissues were significantly high, Fas/FasL mRNA and its protein were up-regulated in lung tissues of lung ischemia/reperfusion injury compared with control (all of P<0.01). The PNS suppressed apoptosis as well as expression of Fas/FasL mRNA and its protein (P<0.05 or P<0.01, respectively). There was a significant correlation between expression of Fas/FasL protein, Fas/FasL mRNA and cell apoptosis (r=0.540，0.658，0.668，0.686；all P<0.01). CONCLUSIONS: Activation of Fas/FasL system and its initiating cell apoptosis of lung tissues may contribute to the pathogenesis of lung ischemia/reperfusion injury. The protective effects of PNS include suppressing the activation of Fas/FasL system and blocking apoptosis in lung tissues in lung ischemia/reperfusion injury.     

Mechanisms of hepatocytic mitochondria damage following septic shock

AIM: To study mechanism of hepatocytic mitochondria damage following septic shock. METHODS: 30 SD rats were randomly divided into three groups: sham operation group, 12 h cecal ligation and puncture (CLP) group and 16 h CLP group. The model of septic shock was made by cecal ligation and puncture. The liver mitochondria respiratory control rate (RCR), phosphate/oxygen (P/O) and ATPase activities were assayed. RESULTS: In 12 h CLP group mean artery pressure (MAP) [(9.54±1.26)kPa] was significantly lower than sham operation group [(14.58±1.32)kPa,P<0.05]. However, mortality was obviously higher than sham operation group (P<0.05), the liver mitochondria respiratory control rate (1.27±0.25), phosphate/oxygen (1.67±0.34) and Na⁺-K⁺-ATPase (40.80±3.45), Ca²⁺-ATPase (58.00±2.43), Mg²⁺-ATPase (78.30±4.16), Ca²⁺-Mg²⁺-ATPase(2.70±2.25) activities decreased strikingly. The difference between 12 h CLP group and sham operation group was significant (P<0.05), 16 h CLP groups was more lower than 12 h CLP group. As RCR, P/O and ATPase activities were significantly reduced, mortality significantly increased. Futhermore, obvious positive correlation was showed between them (r=0.892,P<0.01;r=0.834,P<0.01). CONCLUSION: Liver mitochondria function of ingestion-oxygen and phosphorus-acidification are decreased and membrane fluxion is weaken. Energy metabolism is blocked and Ca²⁺-Mg²⁺ shows imbalanced. All of them cause hepatocytic mitochondria injury following septic shock.     

Inhibition of electrolyte transport in isolated guinea-pig ileum by berberine

AIM: To investigate the effects of berberine (Ber) on ion transport and hypersecretion induced by cholera toxin(ChT) in ileum and its mechanism. METHODS: Ussing Chamber technique was used to measure the potential difference(PD), short-circuit current(SCC), and the resistance(R) in isolated guinea-pig ileum. The effects of Ber on PD, SCC and R in glucose Tyrode solution or glucose-free Tyrode solution were examined. The secretory diarrhoea model was made by ChT to investigate the effects of ChT and ChT+Ber on PD, SCC and R. RESULTS: (1)The PD and SCC were decreased in glucose Tyrode solution by the Ber added in mocosal side or in serosal side in ileum of normal guinea-pig and which was with secretory diarrhoea (P<0. 01, P<0. 01). 1. 0 mmol·L^-1 Ber in serosal face also decreased the R (P<0. 01). (2) In glucose-free Tyrode solution, the Ber in mucosal face did not decrease the PD, SCC and R, but 1. 0 mmol·L^-1 Ber in serosal face decreased the PD, SCC and R (P<0. 01, P<0. 01, P<0. 05). CONCLUSION: Ber not only inhibited electrogenic ion transport in normal guinea-pig ileum, but also reversed hypersecretion induced by ChT.     

Effect of ghrelin on iNOS expression in alveolar macrophages and lung tissues in rats with sepsis-associated lung injury

AIM: To investigate the effect of ghrelin on inducible nitric oxide synthase (iNOS) expression in alveolar macrophages and lung tissues in sepsis-induced acute lung injury (ALI) rats. METHODS: The septic rat model was established by cecal ligation and puncture (CLP). Male SD rats were divided into sham group, CLP group and CLP+ghrelin group. The rats in the former 2 groups were further divided into 3 subgroups, which were 6 h, 12 h and 20 h post-operation groups. Ghrelin was administered by intraperitoneal injection at 3 h and 15 h after operation in ghrelin group. The samples were harvested 20 h after operation. The mRNA expression of iNOS in alveolar macrophages collected from bronchoalveolar lavage was detected by RT-PCR. The protein levels of lung iNOS were measured by Western blotting. The lung pathological examination was performed 20 h after operation. RESULTS: In CLP group, the mRNA expression levels of iNOS in the alveolar macrophages were 1.33±0.05, 1.44±0.08, 1.57±0.11 at 6 h, 12 h and 20 h after CLP, respectively, which were higher than that in sham group, but did not show time correlation. However, it was lower in CLP group than that in CLP+ghrelin group at 20 h after CLP (2.27±0.37, P<0.05). At 20 h after CLP, the protein level of lung iNOS was decreased in CLP+ghrelin group (0.87± 0.03) as compared with CLP group (1.08±0.05). Compared with sham group, the histopathological score was increased in both CLP group and CLP+ghrelin group, but it was lower in CLP+ghrelin group (5.83±0.477) than that in CLP group (7.83±0.75). CONCLUSION: Ghrelin treatment improves the degree of ALI. During 6 h to 20 h after CLP, the mRNA expression of iNOS in alveolar macrophages was elevated, but the difference was not seen as the time went on. Ghrelin up-regulates the mRNA expression of iNOS in alveolar macrophages and inhibits iNOS expression in lungs of septic rats.     

Effects of leptin on hypoxia-reoxygenation induced apoptosis in human L02 cells and expression of Fas and FasL

AIM: To observe the effect of leptin (LEP) on hypoxia-reoxygenation induced apoptosis in L02 cells.METHODS: In the experiment, L02 cell injury was induced by hypoxic air (95%N2 and 5% CO2). The cultured L02 cells were divided into hypoxic 12 h group (IR group) alone, normal control group and the hypoxic plus leptin (100 μg/L, 200μg/L, 400 μg/L, 800 μg/L and 1 600 μg/L) treatment groups in vitro. Flow cytometry, terminal dUTP nick-end labeling (TUNEL) assay and fluorescent quantitative PCR were used to measure the changes of apoptosis in L02 cells and expression of Fas/FasL mRNA.RESULTS: (1) The percentage of L02 cells apoptosis and positive TUNEL cells significantly increased in IR group compared to control group (P<0.01). When L02 cells were treated with LEP, the percentage of cell apoptosis and positive TUNEL cells were decreased compared to IR group. (2) Compared to control group, the Fas/FasL mRNA expression significantly increased in IR group (P<0.01). When L02 cells were treated with LEP, the Fas/FasL mRNA expression decreased compared to IR group, the effect of LEP at concentration of 400 μg/L was obviously (P<0.05). CONCLUSION: LEP decreases the apoptosis of hypoxic-reoxygenation L02 cells by down-regulation of Fas/FasL mRNA expression in L02 cells.     

Effects of Tertram ethylpyrazine on expression of Fas/FasL mRNA in lung tissue with ischemia-reperfusion injury in rabbits

AIM: To study the expression of Fas/FasL mRNA in lung tissue with ischemia-reperfusion lung injury in rabbits and the relationship with the apoptosis,and to observe the effects of Tertram ethylpyrazine on them.METHODS: The pulmonary ischemia-reperfusion models in rabbits with occlusion of left pulmonary hilum for 1 h and then reperfusion 1,3,5 h respectively were used in this experiment.In TMP group,Tertram ethylpyrazine was intravenously dropped at dose of 60 mg/kg at 1 h before ischemia.The TUNEL technique was used to explore apoptotsis of pulmonary cells.In situ hybridization was performed on the rabbit lung tissue to assay the expression of Fas/FasL mRNA.RESULTS: Apoptosis of pulmonary cells was found in both IR group and TMP group.Compared with group IR,the apoptosis index (AI) was decreased obviously in group TMP (P<0.01).There was a significant positive correlation between the expression of Fas/FasL mRNA and the apoptosis of pulmonary cells (r₁=0.900,r₂=0.869,P<0.01).CONCLUSION: The activation of Fas/Fas-L system may contribute significantly to induce pneumocyte apoptosis in pulmonary ischemic injury.Tertram ethylpyrazine inhibits the activation of Fas/FasL system to decrease apoptosis in pulmonary tissue,which may protect the pulmonary tissues in ischemia injury.     

The protective effect of cistanche deserticola Y.C Ma. on thymocytes in septic rats

AIM: To study the protective effect of cistanche deserticola Y. C Ma. on thymocytes in septic rats. METHODS: Cecum ligation perforation (CLP) was used to induce sepsis. Treatment group was treated with cistanche deserticola Y. C Ma. （1.25 g·kg-1·d-1， ig） for 14 days before CLP. Animals were killed 12 h or 24 h after CLP and thmocytes were collected. The ratio of thmocyte apoptosis and mitochondrial membrane potential were determined by the flow cytometry. The ATP activity was detected by spectrophotography. RESULTS: The rate of thmocyte apoptosis significantly increased 12 h after CLP. The ATP activity decreased 24 h after CLP was significant. The extract of desert living cistanche effectively repressed the apoptosis of thymocytes and maintained the mitochondrial membrane potential. CONCLUSIONS: The extract of cistanche deserticola Y. C Ma. protects thymocytes against apoptosis induced by sepsis. Maintaining of mitochondrial membrane potential may be the protective mechanism.     

Establishment of a mouse model of sepsis associated encephalopathy and preliminary research of cognitive dysfunction in this model

AIM: To establish the mouse model of sepsis-associated encephalopathy (SAE) and the preliminary research of cognitive dysfunction in this model. METHODS: SPF male C57BL/6J mice of 8~10 weeks old were selected. The first part of the experiment divided the mice into 4 groups randomly, namely control group, cecal ligation and puncture (CLP)1 group and CLP2 group (CLP was performed with 7 and 12 syringe needle respectively). The mice in sham operation group were only laparotomy. In the second part of the experiment, the mice were randomly divided into control group, sham operation group and CLP group. The Kaplan-Meier method was used to analyze the postoperative survival rate of the mice in the first part experiment. The neurobehavioral scores were used to evaluate the neurobehavioral changes of the mice. The Morris water maze test and the passive avoidance experiment were used to detect the changes of cognitive memory function in the mice. The pole test and the wire suspension test were used to test the motor coordination of the mice. The serum levels of prostaglandin E2 (PGE₂) were measured by ELISA. RESULTS: In the first part of the experiment, the CLP mice showed obvious symptoms such as lethargy, piloerection, chills and anorexia. The 48 h mortality in CLP1 group and CLP2 group were 20% and 30% respectively. In the 2 parts of the experiments, the neurobehavioral scores of the CLP mice were significantly lower than those in control group and the sham operation group (P<0.01). In CLP mice, the escape latency time of the Morris water maze was significantly prolonged (P<0.01), the target quadrant dwell time and the number of crossing platforms were decreased (P<0.01), the scores in the suspension experiment and the pole test were significantly reduced (P<0.01), the activity of the mice was decreased or even did not enter the darkroom in the step-through test (P<0.05). In the second part of the experiment, the serum level of PGE₂ in the mice after CLP was significantly increased (P<0.01). CONCLUSION: A stable mouse model of sepsis-associated encephalopathy is successfully established by cecal ligation and puncture with 12 syringe needle. The SAE mouse model established by this method is useful for investigating the learning and memory cognitive dysfunction.     

Berberine promotes anti-proliferative effects of doxorubicin in T24 bladder cancer cells by enhancing apoptosis

AIM: To observe the effect of berberine (Ber) on doxorubicin (DOX)-induced apoptosis in bladder cancer T24 cells. METHODS: The cells were exposed to DOX in the presence or absence of different concentrations of Ber. The viability of the cells was determined by CCK-8 assay. The apoptosis was measured by Hoechst 33258 staining and the protein levels of cleaved caspase-3, cleaved caspase-9, Bcl-2 and Bax were detected by Western blotting.RESULTS: Ber enhanced the inhibitory effect of DOX on the viability of T24 cells and promoted DOX-induced apoptosis in T24 cells. DOX increased the protein levels of cleaved caspase-3, cleaved caspase-9 and Bax, all of which were enhanced by treatment with Ber. In contrast, Ber exposure further decreased the expression of Bcl-2 in DOX-treated T24 cells.CONCLUSION: Ber enhances the anti-proliferative effects of DOX through promoting apoptosis in bladder cancer cells.     

Effect of Fas/FasL on late reperfusion of acute myocardial infarction and the potential oxidative stress mechanism in canine

AIM: To discuss the effect of Fas/FasL on the late reperfusion of acute myocardial infarction (AMI) and the potential oxidative stress mechanism. METHODS:Eighteen anesthetized dogs were randomly divided into three groups: late reperfusion group (n=6): ligated the coronary for 6 h, followed by reperfusion for 6 h; permanent ischemia group (n=6): after pericardium were opened for 6 h, ligated the coronary for 6 h, and did not reperfuse; control group (n=6): did not ligate the coronary but operation last for 12 h. Infarction brim myocardial Fas/FasL was detected by immunohistochemistry. Apoptosis index (AI) was detected by TUNEL. SOD and GR activity and MDA content were detected by colorimetry. RESULTS:The expression of Fas/FasL and apoptosis index were significantly higher in permanent ischemia group and late reperfusion group than those in control group (P<0.05, P<0.01), and the difference between them was also significant (P<0.05). SOD and GR activities were lower in permanent ischemia group and late reperfusion group than those in control group (P<0.05, P<0.01). The MDA contents in permanent ischemia group and late reperfusion group were higher than that in control group (P<0.05). CONCLUSION:The late reperfusion of AMI promotes the expression of Fas/FasL and myocardial apoptosis, and it may be due to oxidative stress mechanism.     

Heme oxygenase-1 protects renal function in septic rats via influencing expression of thrombomodulin

AIM: To investigate the protective effect of heme oxygenase-1 (HO-1) on the kidney of septic rats and the influence of HO-1 on the expression of thrombomodulin (TM) in the kidney. METHODS: Sepsis in rats was developed with cecal ligation and puncture (CLP). The septic rats were randomly divided into sham group, CLP group, CLP+HO-1 inducer group and CLP+HO-1 inhibitor group (n=18). The plasma levels of creatinine (Cr), cystatin-C (Cys-C), carboxyhemoglobin (COHb), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and TM, and the changes of prothrombin time (PT) and activated partial thromboplastin time (APTT) in each group were measured. Histopathological examination was performed in the kidney. The expression of TM in the kidney tissue was detected by Western blot. RESULTS: Compared with sham group, significantly elevated plasma levels of Cr, Cys-C, TNF-α, IL-1β and TM (P<0.05), shortened PT and APTT (P<0.05), significantly increased microthrombus formation, and lowered TM expression in the kidney (P<0.05) of CLP group were observed. The administration of hemin lowered the plasma levels of Cr, Cys-C, TNF-α, IL-1β and TM (P<0.05), prolonged PT and APTT (P<0.05), attenuated microthrombus formation, and up-regulated the expression of TM in the kidney (P<0.05). In contrast, ZnPP had the opposite effects. CONCLUSION: HO-1 increases the expression of TM in the kidney and exerts anticoagulatory and antiinflammatory effects, thereby improving renal function in the septic rats.     

Relation of Fas/FasL-mediated caspase-3 activation with acinar cell apoptosis in rats with acute pancreatitis

AIM: To observe the expression and interrelationship of apoptosis controlling genes and proteins Fas, FasL, caspase-3 in rats with acute pancreatitis (AP). METHODS: Forty male Sprague Dawley rats were randomly divided into four groups, 10 rats each group. Acute pancreatitis with different inflammatory degree was induced by retroinjecting 2.0%, 3.5% or 5.0% sodium taurocholate at dose of 1 mL/kg body weight into the pancreaticobiliary duct. All the rats were sacrificed 6 h after operation. The pathologic changes of pancreas were observed under optical microscope. The protein and mRNA expressions of Fas, FasL, caspase-3 in pancreatic tissue were measured by Western blotting and RT-PCR. The apoptosis of acinar cells was measured by the methods of in situ end labeling. RESULTS: In normal pancreatic tissue, there appeared the protein and mRNA expression of Fas, FasL, caspase-3. In acute pancreatitis with different inflammatory degree, with the degree of inflammation worsen, the apoptosis cells tapered, the expression of above protein and mRNA also descended gradually. Furthermore, the variation tendency of caspase-3 among the four groups was in coincidence with Fas or FasL. CONCLUSION: Fas/FasL -mediated apoptotic pathway participates in the regulation of acinar cell apoptosis in acute pancreatitis.     

Effect of respiratory syncytial virus on apoptosis and expressions of FasL,Fas, Bcl-2 and Bax

AIM: To study the relationship between respiratory syncytial virus (RSV) infection and apoptosis, between RSV infection and expressions of FasL, Fas, Bcl-2 and Bax. METHODS: Apoptotic cells were examined by flow cytometry and transmission electron microscope. Immunohistochemical analysis was used to detect the expressions of apoptosis-associated gene FasL, Fas, Bcl-2 and Bax in A549 cells during RSV infection. RESULTS: Apoptotic index increased at 72 h and 120 h postinfection. Apoptotic cells were detected by transmission electron microscope. High-expressions of FasL, Fas and Bax genes and low-expression of Bcl-2 gene were detected by immunohistochemical staining. CONCLUSION: Apoptosis in A549 cells was induced by RSV infection. This apoptosis may be induced by up-regulating the expression of FasL, Fas, Bax genes and down-regulating the expression of Bcl-2 gene.     

Protective effects of berberine in pancreatic islet beta cells

AIM: To explore the protective effects of berberine （Ber） on islet beta cells and related possible mechanisms. METHODS: The injury of INS-1 cells was induced by treatment with alloxan (Axn). Berverine was then given at serial concentrations. The cells were divided into control group (Con), injury group (Axn), low-dose berberine group (LBer), medium-dose berberine group (MBer) and high-dose berberine group (HBer). Flow cytometry was employed to detect the apoptosis. The activation of PTEN/PI3K/Akt and HNF-1α/PDX-1 pathways and the protein level of cleaved caspase-3 were evaluated by Western blotting. The insulin releases under normal or high-glucose stimulation were measured by ELISA. RESULTS: Compared with Con group, the apoptotic rate increased significantly in Axn group. Berberine treatment reduced the apoptotic rate in LBer group, MBer group and HBer group in a concentration-dependent manner. Compared with Con group, the protein levels of PTEN and cleaved caspase-3 increased, while PI3K and phosphorylation of Akt decreased significantly in Axn group. However, this effect was reversed by berberine in a concentration-dependent manner. Compared with Con group, the activation of HNF-1α/PDX-1 signaling pathway was inhibited in Axn group but recovered by berberine administration. The abilities of releasing insulin under normal or high-glucose stimulation were impaired in Axn group but recovered by berberine treatment in LBer group, MBer group and HBer group in a concentration-dependent manner.CONCLUSION: Berberine shows protective effects against alloxan-induced damage in beta cells by inhibiting apoptosis and recovering insulin secretion, thus attenuating the activation of PTEN/PI3K/Akt and HNF-1α/PDX-1 signaling pathways.