Neuroprotective effects of Ginkgo biloba dropping pills in Parkinson's disease / 药物分析学报

Parkinson's disease(PD)is the second most common neurodegenerative disease in the world;however,it lacks effective and safe treatments.Ginkgo biloba dropping pill(GBDP),a unique Chinese G.biloba leaf extract preparation,exhibits antioxidant and neuroprotective effects and has a potential as an alternative therapy for PD.Thus,the aims of this study were to evaluate the effects of GBDP in in vitro and in vivo PD models and to compare the chemical constituents and pharmacological activities of GBDP and the G.biloba extract EGb 761.Using liquid chromatography tandem-mass spectrometry,46 GBDP constitu-ents were identified.Principal component analysis identified differences in the chemical profiles of GBDP and EGb 761.A quantitative analysis of 12 constituents showed that GBDP had higher levels of several flavonoids and terpene trilactones than EGb 761,whereas EGb 761 had higher levels of organic acids.Moreover,we found that GBDP prevented 6-hydroxydopamine-induced dopaminergic neuron loss in zebrafish and improved cognitive impairment and neuronal damage in methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced PD mice.Although similar effects were observed after EGb 761 treatment,the neuroprotective effects were greater after GBDP treatment on several endpoints.In addition,in vitro results suggested that the Akt/GSK3β pathway may be involved in the neuroprotective effects of GBDP.These findings demonstrated that GBDP have potential neuroprotective effects in the treatment of PD.

Effect of Xihuang pills on apoptosis and expression of ER and PR in rat mammary epithelial cells / 中国药理学通报

Aim To investigate the effect of Xihuang pills on estradiol (E2) and progesterone (P)-induced apoptosis of rat mammary epithelial cells and the expression of estrogen and progesterone receptors, and to explore its anti-mammary hyperplasia mechanism. Methods Rat mammary epithelial cells were treated with different concentrations of Xihuangpills containing serumafter E2, P induced proliferation. The cell proliferation was detected by CCK-8 method (24, 48, 72 h). The apoptosis was detected by flow cytometry after AnnexinV-FITC/PI double staining. The average optical density of Bax and Bcl-2 was detected by immunohistochemistry; Estradiol receptor alpha (ER-α), estradiol receptor beta (ER-β), progesterone receptor (PR) protein expression were detected by Western blot. Results The serum containing Xihuang pillssignificantly inhibitedthe proliferation of rat mammary epithelial cells after E2and P treatment (P < 0. 05), and induced apoptosis, affecting the expression of apoptosis-related proteins Bcl-2 and Bax (P < 0. 05), and effectively inhibiting the expression of estrogen receptors ERα, ERβ. Conclusions Xihuang pills can induce the apoptosis of mammary epithelial cells by regulating related apoptotic proteins, and regulate the secretion of estrogen and progesterone in breast tissues, affecting the proliferation and rejuvenation of breast, which is of great significance for the treatment of breast hyperplasia.

Miltirone induces cell death in hepatocellular carcinoma cell through GSDME-dependent pyroptosis

Pyroptosis is a form of programmed cell death, and recently described as a new molecular mechanism of chemotherapy drugs in the treatment of tumors. Miltirone, a derivative of phenanthrene-quinone isolated from the root of Bunge, has been shown to possess anti-cancer activities. Here, we found that miltirone inhibited the cell viability of either HepG2 or Hepa1-6 cells, and induced the proteolytic cleavage of gasdermin E (GSDME) in each hepatocellular carcinoma (HCC) cell line, with concomitant cleavage of caspase 3. Knocking out switched miltirone-induced cell death from pyroptosis to apoptosis. Additionally, the induction effects of miltirone on GSDME-dependent pyroptosis were attenuated by siRNA-mediated caspase three silencing and the specific caspase three inhibitor Z-DEVD-FMK, respectively. Miltirone effectively elicited intracellular accumulation of reactive oxygen species (ROS), and suppressed phosphorylation of mitogen-activated and extracellular signal-regulated kinase (MEK) and extracellular regulated protein kinases 1/2 (ERK1/2) for pyroptosis induction. Moreover, miltirone significantly inhibited tumor growth and induced pyroptosis in the Hepa1-6 mouse HCC syngeneic model. These results provide a new insight that miltirone is a potential therapeutic agent for the treatment of HCC GSDME-dependent pyroptosis.

Evaluation of lymphocyte apoptosis in patients with oral cancer

Abstract Objectives To evaluate apoptotic levels of peripheral blood mononuclear cells (PBMCs) and apoptotic regulatory proteins (Bax and Bcl-2) in lymphocyte subsets of oral cancer (OC) patients and healthy controls (HC). Methodology The percentage of apoptotic cells and lymphocyte counts were measured in the first cohort using PBMCs obtained from 23 OC patients and 6 HC. In the second cohort, (OC, 33; HC, 13), the mean fluorescence intensity (MFI) of Bax and Bcl-2 in CD19+ B, CD4+ T, CD8+ T, and CD16+56+ natural killer (NK) cells was determined via flow cytometry. Results The percentage of apoptotic cells was higher in the PBMCs of OC patients than in HC patients, particularly in patients with stage IV cancer (p<0.05). However, lymphocyte counts were significantly lower in stage IV patients (p<0.05). NK CD19+ B and CD16+56+ cell counts were significantly lower in OC patients compared with HC patients (p<0.001 and p<0.01, respectively), but CD4+ T cells were interestingly significantly higher in OC patients (p<0.001). While Bax MFI was slightly higher, Bcl-2 MFI was significantly lower for all four lymphocyte subsets in OC samples, particularly in stage IV patients, when compared with HC. Consequently, Bax/Bcl-2 ratios showed an upward trend from HC to OC patients, particularly those in stage IV. We found similar trends in Bax and Bcl-2 MFI for tumor stage, tumor size, and lymph node involvement. Conclusions The increased lymphocyte apoptosis in stage IV OC patients may be related to higher Bax levels and lower Bcl-2 levels. The Bax/Bcl-2 ratio in lymphocytes may be useful to determine the prognosis of OC patients, and could be considered a mean for supportive treatment in the future.

Acupuncture of intraorbital and extraorbital acupoints reduces apoptosis of retinal ganglion by down-regulating expression of Caspase-3 and ratio of Bax/Bcl-2 in rabbits with nonarteritis anterior ischemic optic neuropathy / 针刺研究

OBJECTIVE: To observe the effect of acupuncture of intraorbital and extraorbital acupoints on apoptosis and expression of Bax, Bcl-2 and Caspase-3 proteins of retinal ganglion tissue in rabbits with nonarteritis anterior ischemic optic neuropathy (NAION), so as to reveal its mechanism underlying improvement of NAION. METHODS: Female New Zealand rabbits were randomly divided into four groups: model, intraocular needling (ION), extraocular needling (EON), ION＋EON groups (n＝5 per group), and the 5 healthy eyes of those rabbits in the model group were selected to be used as the control group. The NAION model of the right eye was established by photodynamic stroke method. For ION group, acupoints "Jingming" (BL1), "Chengqi" (ST1) and "Qiuhou" (EX-HN7) were needled, and for EON group, "Cuanzhu" (BL2), "Yuyao" (EX-HN4) and "Qiaoming" (EX) were needled with filiform needles, followed by retaining the needles for 30 min. For ION＋EON group, the 6 acupoints were needled simutaneously. The treatment was conducted once daily for 3 days. The apoptosis of retinal ganglion tissue was detected by using TUNEL fluorescence labeling, and the expression of Bax, Bcl-2 and active Caspase-3 in the retinal ganglion were detected by immunohistochemistry. RESULTS: Following modeling and compared with the control group, the number of the apoptotic cells, and the expression levels of Bax and Caspase-3 proteins, as well as the ratio of Bax/Bcl-2 were significantly increased in the model group (P0.05). CONCLUSION: Acupuncture of intraorbital and extraorbital acupoints can reduce apoptosis of retinal ganglion in NAION rabbits via inhibiting the activation of Caspase-3 protein and ratio of Bax/Bcl-2, and up-regulating the expression of Bcl-2 protein.

Low spontaneous apoptosis index at diagnosis predicts a high-risk phenotype in paediatric acute lymphoblastic leukaemia

Background & objectives: Significance of apoptosis as a prognostic marker is less well studied in paediatric acute lymphoblastic leukaemia (ALL) cases. Hence, a prospective study, involving 30 paediatric ALL cases, was done to assess the clinical relevance of in vivo apoptosis. Methods: Peripheral blood mononuclear cells from all patients were subjected to annexin V/propidium iodide staining to detect the degree of apoptosis [apoptotic index (AI)] at day 0 and day 35 post-induction chemotherapy. In addition, Bax and Bcl2 apoptotic protein expressions were studied at day 0 and their relative fluorescence mean intensity (RFMI) ratios were calculated. Results: Mean age of patients was 5.1 years. Of the 30 cases, 21 (70%) were at standard-risk, five (17%) at intermediate and four (13%) at high risk. Majority (83%) were B-ALL. Day 8 absolute blast count was >1000/?l in seven (23%) and <1000/?l in 23 of 30 (77%) cases. Day 35 marrow was M1 in 23 (92%) and M2 in two of 25 (8%) cases. AI at day 0 and day 35 ranged from 0.9 to16.6 per cent and 1.4 to 62.8 per cent with a mean of 5.90 and 19.64 per cent, respectively. The Bax/Bcl2 ratio ranged from 0.2 to 3.5 with a mean of 0.83. The ratio was predominantly anti-apoptotic, i.e. <1 (77%). A significant association was noted between low AI at day 0 and high total leucocyte count (P=0.02), T-cell phenotype (P=0.043) and high-risk as per NCI category (P=0.025). Significant increase (>30%) in day 35 AI was seen in only six cases. Interpretation & conclusions: Our study showed that low AI at day 0 was associated with a high-risk clinical phenotype in paediatric ALL. However, studies on larger group, especially with longer follow up or study of relapse cases, will help draw conclusions regarding apoptosis assessment in paediatric ALL.

The effect of Ulinastatin on myocardial cell apoptosis and Caspase-3 channel / 中华急诊医学杂志

Objective To investigate the effect of ulinastatin on myocardial injury in rats with sepsis.Methods Thirty male SD rats were randomly (random number) divided into 3 groups:sham operation group (Sham group,n=10),sepsis group (sepsis group,n=10) and ulinastatin group (UTI group,n=10).The sepsis model of the rats was subjected to cecal ligation and puncture (CLP).Rats of UTI group were given 200 000 U/kg ulinastatin at 6 hour after modeling,and dosing was repeated every 12 h.Blood samples were collected from inferior vena cava at 6,12,24,36 h after modeling for determination of cardiac troponin-Ⅰ (cTnI) and the inflammatory factor by ELISA,then the rats were sacrificed and hearts were removed for myocardial tissue stained with hematoxylin eosin (HE) staining,the expression of Bax/Bcl-2 protein level and myocardial cell apoptosis were detected by TUNEL.The level of caspase-3 protein in myocardial tissue was detected by Western-blot.Results The level of cTnI (ng/mL) in serum in UTI group at 6,12,24 and 36 h after modeling were significantly lower compared with sepsis group(P＜0.05).The protein expression levels of TNF-α and IL-6 (ng/mL) in group UTI were higher than those in Sham group,but was significantly lower than those in Sepsis group (P＜0.05).HE staining showed that inflammatory cell infiltration present in myocardial cells,edema and vacuole formation were observed in sepsis group,while those were significantly attenuated in UTI group compared with sepsis group.UTI increased the level of myocardial Bcl-2 protein in the rats (P＜0.05),and reduced the level of myocardial Bax protein (P＜0.05).TUNEL and HE staining showed apoptosis cells in UTI group was significantly reduced compared with sepsis group [(32.2±4.8)％ vs.(58.4±5.6)％,P＜0.05];Western-blot method showed the level of Caspase-3 protein in UTI group was higher than that in group sham (0.32±0.048) vs.(0.12±0.03),P＜0.05],but significantly lower than that of Sepsis group [(0.32±0.048) vs.(0.55±0.052),P＜0.05].Conclusions Ulinastatin can reduce proinflammatory mediators release in the blood of sepsis rats,and inhibit the apoptosis of myocardial cells protecting myocardial cells through the regulation of the Caspase-3 pathway during sepsis.

Anti-proliferative Effects of Androctonus amoreuxi Scorpion and Cerastes cerastes Snake Venoms on Human Prostate Cancer Cells

The present study evaluated the effects of Androctonus amoreuxi scorpion venom, Cerastes cerastes snake venom and their mixture on prostate cancer cells (PC3). An MTT assay was used to determine the anti-proliferative effect of the venoms, while quantitative real time PCR was used to evaluate the expression of apoptosis-related genes (Bax and Bcl-2). Furthermore, colorimetric assays were used to measure the levels of malondialdehyde (MDA) and antioxidant enzymes. Our results show that the venoms significantly reduced PC3 cell viability in a dose-dependent manner. On the other hand, these venoms significantly decreased Bcl-2 gene expression. Additionally, C. cerastes venom significantly reduced Bax gene expression, while A. amoreuxi venom and a mixture of A. amoreuxi & C. cerastes venoms did not alter Bax expression. Consequently, these venoms significantly increased the Bax/Bcl-2 ratio and the oxidative stress biomarker MDA. Furthermore, these venoms also increased the activity levels of the antioxidant enzymes, catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase. Overall, the venoms have cytotoxic and anti-proliferative effects on PC3 cells.

Study on the drug-resistant reversal effects of ginsenoside Rh2 in human hepatocellular carcinoma HepG2/ADM cells and its mechanism / 医学研究生学报

Objective Ginsenoside Rh2 can inhibit the proliferation of a variety of malignant tumor cells.However, little research has been done on the sensitivity of Rh2 in human hepatocellular carcinoma HepG2/ADM cells with multidrug resistance(MDR).This study aimed to explore the reversing effects of ginsenoside Rh2 on the MDR of human hepatocellular carcinoma HepG2/ADM cells and its potential mechanism.Methods MTT assay was applied to detect the effect of Rh2(0-250 μg/mL) on the viability of HepG2/ADM cells and screen out the optimum drug-resistant reversal concentration of Rh2.Cells were divided into 3 groups: HepG2/ADM group (without any medicine treatment), ADM group(ADM treatment for 48 h), ADM+40 μg/mL Rh2 group (pretreatment of 40μg/mL Rh2 for 30 min followed by ADM treatment for 48 h).Flow cytometry was applied to detect the effect of Rh2 on the fluorescence intensity of cellular Rh-123.RT-PCR was used to measure the expression of MDR1 gene.Western blot was used to detect the protein levels of P-gp, Bax, Bcl-2 and cleaved caspase-3.Results 40 μg/mL ginsenoside Rh2 significantly reversed the MDR of HepG2/ADM cells by a 2.55-to-3.70-fold increase in sensitivity.Furthermore, compared with ADM group, the efflux of Rh-123 in HepG2/ADM cells were remarkably inhibited by Rh2 in ADM+40 μg/mL Rh2 group (65.83±1.78 vs 78.21±1.26, P<0.01), along with the down-regulated expressions of MDR1 (0.48±0.02 vs 0.86±0.05, P<0.05), P-gp(0.97±0.04 vs 1.91±0.03,P<0.01), Bcl-2(1.25±0.05 vs 1.86±0.03, P<0.05) and the up-regulated protein level of Bax (1.76±0.04 vs 1.25±0.02,P<0.05) and cleved caspase-3(0.42±0.04 vs 38.26±5.45,P<0.05).Conclusion Ginsenoside Rh2 can effectively reverse the MDR of HepG2/ADM cells, and the potential mechanism is related to the decreased expressions of MDR1 and P-gp, the increasing drug concentration inside the cells and the Bax/Bcl-2 signaling pathway.

Study effects and mechanism of PHB on reactive oxygen species and apoptosis in cardiomyocytes under high glucose / 中国免疫学杂志

Objective:To investigate the effect and mechanism of PHB on the apoptosis of H 9C2 cardiomyocytes under high glucose.Methods:Select pCDNA3-PHB and pCDNA3-NC recombinant plasmid transfect into cardiomyocytes and effected by high glu-cose.The experiment were divided into four groups:control group,high glucose group(HG group),high glucose+empty vector group (HG+pCDNA3-NC group),high glucose+pCDNA3-PHB plasmid group(HG+pCDNA3-PHB group).The expression of PHB,Bax/Bcl-2,c-caspase3,AKt and p-AKt protein after transfected were detected by Western blot .ROS were detected under high glucose by DHE staining.The TNF-αmRNA and IL-6 mRNA were detected by Real-time quantitative PCR(qRT-PCR).Flow cytometry was used to detect the apoptotic rate .Results:The recombinant plasmid pCDNA 3-PHB could increase the expression of PHB .The apoptotic rate of HG group,HG+pCDNA3-NC group and HG+pCDNA3-PHB group were significantly higher than control group (P<0.05).Compared with HG group,the apoptotic rate of HG+pCDNA3-NC group was not significant(P>0.05),but that of HG+pCDNA3-PHB group was significantly lower(P<0.05).The levels of ROS,TNF-αmRNA,IL-6 mRNA,Bax/Bcl-2 and c-caspase3(P<0.05) protein in the HG group,HG+pCDNA3-NC group,HG+pCDNA3-PHB group were significantly higher than the control group .Compared with HG group , the level of ROS,TNF-αmRNA,IL-6 mRNA,Bax/Bcl-2 and c-caspase3 (P>0.05)protein in the HG+pCDNA3-NC group were no significant difference,but the ROS,TNF-αmRNA,IL-6 mRNA、Bax/Bcl-2 and c-caspase3 (P<0.05)protein in the HG+pCDNA3-PHB group were significantly decreased .Statistical analysis showed that there was no significant difference of AKt protein in each group (P>0.05).The relative expression of p-AKt protein in HG group,HG+pCDNA3-NC group and HG+pCDNA3-PHB group were signif-icantly lower than that in control group ( P<0.05 ) ,HG+pCDNA3-NC group was no significantly different than HG group ( P>0.05 ) , However ,HG+pCDNA3-PHB group was significantly higher than HG group ( P<0.05 ) .Conclusion: PHB overexpression can inhibit the apoptosis of H9C2 cardiomyocytes in high glucose environment ,andreduce the inflammatory response by effected the regulation of PI3K/Akt signaling pathway,reduced ROS,Bax/Bcl-2,c-caspase3 protein levels,that give us a new ideas and methods of DCM .

Involvement of p38-p53 signal pathway in neuroprotective effects of DGMI on SH-SY5 Y cells damaged by oxygen-glucose deprivation / 中国药理学通报

Aim To investigate the protective effects of Diterpene Ginkgolides Meglumine Injection(DGMI)on SY5 Y cells damaged by oxygen-glucose deprivation and its functional mechanisms.Methods After 4 h of OGD,the cells were treated with 25 mg·L-1 drugs for 1 h.Subsequently,cell viabilities were measured by cell counting kit-8(CCK-8 kit)and cell apoptosis was measured by flow cytometric analysis.Furthermore, the mitochondrial membrane potential was detected by rhodamine123 staining.The levels of phospho-p38, phospho-p53,Bcl-2,Bax and cleaved caspase-9/3 were evaluated by western blot.Results DGMI signif-icantly increased the cell viabilities of SY5 Y cells dam-aged by OGD,and reduced OGD-elicited dissipation of mitochondrial membrane potential and cell apoptosis. Furthermore,DGMI also reduced p-p38,p-p53,Bax/Bcl-2 ratio,cleaved caspase-9 and cleaved caspase-3. Conclusion DGMI shows good neuroprotective effects on SY5 Y cells after oxygen-glucose deprivation.The underlying mechanisms may be associated with the sup-pression of p38/p53/Bcl-2 /caspase-9/caspase-3 sig-naling pathway.

Effect of gambogic acid on cell apoptosis and expressions of Bax, Bcl-2 and Caspase-3 in colorectal cancer cells with / 实用医学杂志

Objective To investiagate cell apoptosis and expressions of Bax,Bcl-2 andCaspase-3 in gambogic acid-treated colorectal cancer cells. Methods SW480/LOVO colorectal cancer cells were treated by gambogic acid. Cell Counting Kit-8 assay (CCK-8) was used to test cell proliferation. Microscopy was used to check the morphological changes. Immunofluorescence staining technique was used to detect cell apoptosis. Expressions of Bax,Bcl-2 and Caspase-3 protein were detected by Western blot assay. Results Gambogic acid inhibited the proliferation of SW480/LOVO in a dose and time-dependent manner. Gambogic acid could induce cell apoptosis. Gambogic acid increased expressions of Caspase-3 and Bax, increased the ratio of Bax/Bcl-2, and decreased Bcl-2 protein expression. Conclusion Gambogic acid can inhibit proliferation and induce apoptosis of SW480LOVO cells, with the mechanism of up-regulation of Bax/Bcl-2 and activation of Caspase-3.

Inhibitory effect of deoxyschizandrin on growth of brain glioma cells and its mechanism / 吉林大学学报(医学版)

Objective:To study the inhibitory effect of deoxyschizandrin on the growth of brain glioma C6 cells, and to explore its mechanism.Methods:The rat glioma C6 cells were cultured and divided into control group,50, 100,and 200 mg·L-1 deoxyschizandrin groups.The proliferation rates of C6 cells were examined by MTT assay;the changes of cell cycles were examined by flow cytometry;the expression levels of CyclinD1,Bax,Bcl-2 and Caspase-3 proteins in supernant were detected by ELISA assay. Results:Compared with control group, the proliferation rates at 24 and 48 h in 50,100,and 200 mg·L-1 deoxyschizandrin groups were significantly decreased (P <0.01),and the proliferation rates at 72 h in 100 and 200 mg·L-1 deoxyschizandrin groups were significantly decreased (P < 0.05 or P < 0.01 ). Compared with control group, the percentage of cells at SubG1 phase in 200 mg·L-1 deoxyschizandrin group was increased (P < 0.05 ), and the percentage of cells at S phase was decreased (P <0.05).Compared with control group,the expression levels of CyclinD1 in 100 and 200 mg· L-1 deoxyschizandrin groups were decreased (P < 0.01 );the expression levels of Bax protein in deoxyschizandrin groups were increased (P < 0.05 or P < 0.01 ), and the expression level of Bcl-2 protein in 200 mg · L-1 deoxyschizandrin group was decreased (P < 0.01 ), and the Bax/Bcl-2 value in deoxyschizandrin groups were increased (P < 0.01 ); the expression level of Caspase-3 protein in 200 mg · L-1 deoxyschizandrin group was increased (P < 0.01 ).Conclusion:Deoxyschizandrin could inhibit the growth of glioma cells through down-regulating the expression levels of CyclinD1 protein and up-regulating the expression levels apoptotic factors Bax and Bcl-2.

Study on inhibition of Dendrobium officinale extract on gastric carcinogenesis / 中草药

Objective: To explore the inhibitory effects of Dendrobium officinale extract (DOE) on gastric carcinogenesis and its related molecular mechanism. Methods: The rats were randomly divided into normal group, model group, positive drug group, low-dose and high-dose DOE groups. The gastric carcinogenesis model was induced by ig giving 200 mg/kg N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) at doses of 200 mg/kg every 10 days for 3 months. Meanwhile, the rats in the treatment groups were given the corresponding doses of drug while the rats in the control group were given normal saline. At the end of the experiment, the degree of gastric tissue carcinogenesis was calculated by HE staining analysis. The expression levels of EGF, EGFR, Bax, and Bcl-2 mRNA were detected by RT-PCR test in gastric tissue. The levels of EGF and EGFR in plasma were detected by ELISA. Apoptosis was detected by TUNEL assay. Results: Compared with the model group, the high-dose DOE could inhibit the degree of carcinogenesis significantly (P < 0.01). Both high-dose and low-dose DOE could significantly reduce the levels of EGF, EGFR mRNA, and Bcl-2 mRNA, and increase the levels of Bax mRNA in gastric tissue, with statistical significance compared with the model group (P < 0.05, 0.01). In addition, they also reduced the levels of EGF and EGFR in plasma significantly and induced apoptosis. Conclusion: DOE could inhibit the gastric carcinogenesis, which might be related to effects of decreasing the expression of EGF and EGFR mRNA in gastric tissue and EGF and EGFR in plasma and inducing the apoptosis through reducing the expression of Bcl-2 mRNA and increasing Bax mRNA.

Effect of Schisandra lignans on the apoptosis of SHG-44 glioma neurospheres / 中国药学杂志

OBJECTIVE: To explore the mechanism of Schisandra lignans for inducing the apoptosis of human glioma SHG-44 neurospheres. METHODS: The effects of Schisandra lignans on the SHG-44 neurospheres proliferation were detected by MTT. The apoptotic rate was analyzed by Annexin-V/PI double staining. The secretion of the bax, bcl-2 and caspase 3 protein was detected by Elisa assay. The expression of bcl-2 protein was examined by Western blot. RESULTS: Schisandra lignans could inhibit the proliferation of SHG-44 neurospheres with the concentration of 50,100,200 mg · L-1 in a dose-dependent manner. Schisandra lignans could induce the apoptosis of SHG-44 neurospheres in a dose-dependent manner. The secretion of bcl-2 was decreased, but ratio of bax/bcl-2 was increased, caspase 3 was increased, the expression of bcl-2 protein was down-regulated. CONCLUSION: Schisandra lignans could induce the apoptosis of human glioma SHG-44 neurospheres. Its mechanism is correlated with down-regulation of the expression of bcl-2 and up-regulation of the ratio of bax/bcl-2, and then activated caspase 3.

Effect of Electroacupuncture on Learning and Memory Ability of Cerebral Ischemia/Reperfusion Injured Rats / 中国康复理论与实践

@#Objective To investigate the effect of electroacupuncture at Shenting (DU24) and Baihui (DU20) on learning and memory ability, as well as the nerve cell and the expression of mRNA of Bcl-2 and Bax in the hippocampus of rats with cerebral ischemia/reperfusion (I/R) injury. Methods 45 male Sprague-Dawley rats were randomly diveded into sham group (n=15), control group (n=15) and electroacupuncture group (n=15). The latter 2 groups were modeled with middle cerebral artery occlusion for 2 h and reperfusion. The electroacupuncture group received electroacupuncture at Shenting and Baihui for 7 days. They were tested with Morris water maze, and then, their hippocampus was observed under Nissl staining and the level of Bcl-2 and Bax mRNA were determined with RT-PCR. Results The escape latency decreased (P<0.001) and the frequence the platform was crossed increased (P=0.001) in the electroacupuncture group compared with the model group, with less damage of the nerve cell and the expression of Bax mRNA, and more of Bcl-2 mRNA (P<0.05). Conclusion Electroacupuncture can prevent learning and memory ability from cerebral ischemia/reperfusion injury in rats, which may associate with adjustment of expression of apoptosis-related gene to inhibit apoptosis of nerve cell.

Ziyuglycoside II-induced apoptosis in human gastric carcinoma BGC-823 cells by regulating Bax/Bcl-2 expression and activating caspase-3 pathway

Ziyuglycoside II is an active compound of Sanguisorba officinalis L. that has anti-inflammation, antioxidation, antibiosis, and homeostasis properties. We report here on the anticancer effect of ziyuglycoside II on human gastric carcinoma BGC-823 cells. We investigated the effects of ziyuglycoside II on cell growth, cell cycle, and cell apoptosis of this cell line. Our results revealed that ziyuglycoside II could inhibit the proliferation of BGC-823 cells by inducing apoptosis but not cell cycle arrest, which was associated with regulation of Bax/Bcl-2 expression, and activation of the caspase-3 pathway. Our study is the first to report the antitumor potential of ziyuglycoside II in BGC-823 gastric cancer cells. Ziyuglycoside II may become a potential therapeutic agent against gastric cancer in the future.

Apoptotic Activities in Soft Tissue Sarcoma: Immunohistochemical Study and Their Association with Tumour Characteristics

Background: Many studies on the role of apoptosis in cancer development and management have been undertaken. Apoptotic activity depends partly on the balance between anti-apoptotic (Bcl-2) and pro-apoptotic (Bax) activities. This study compared Bcl-2 and Bax expression in the tumour cells and endothelial cells of tumour blood vessels in soft tissue sarcoma, and examined the association of these with tumour characteristics. Methods: A cross sectional (retrospective) study was conducted on 101 cases of various types of soft tissue sarcoma tumour cells and endothelial cells of tumour blood vessels. The immunohistochemical expressions of Bcl-2 and Bax were compared by correlating them according to site, size, depth, tumour margin, lymph node involvement, and histological type. Results: Higher Bax than Bcl-2 expression in tumour cells was observed, although the difference was not statistically significant. There was a significant direct association between Bcl-2 and Bax in tumour cells with endothelial cells. Among tumour characteristics, the only significant correlation was that of the Bcl-2 expression in tumour cells with tumour histological subtypes (synovial sarcoma and leiomyosarcoma). Conclusion: The findings in this study support the role of endothelial cells in the survival and regression of tumour cells in tumour genesis. Therefore, inhibition of endothelial cell survival and activation, or induction of tumour cell apoptosis offers a promising prospect for tumour management.

Effect of tetrandrine eye drops on expression of apoptotic protein and transforming growth factor-β2 in human Tenon capsule fibroblast in vitro / 中华实验眼科杂志

BackgroundOur previous study determined that tetrandrine (Tet) has an inhibitory effect on the proliferation of human Tenon capsule fibroblasts ( TCFs ) in vitro,but its mechanism is poorly understood.ObjectiveThis study was to investigate the effect mechanism of Tet on human TCFs.MethodsHuman TCFs were isolated and cultured from scleral tissue of donor using explant technique.The cells were identified by vimentin antibody staining and morphology.The third generation of cells were seeded in the culture plate at the density of 1 × 105 cells/ml.Twenty-four hours after inoculation,the Tet of 1 × 10-5 mol/L was added in the well of culture plate,and the cells cultured only in 1640 medium served as the control group.The apoptosis of the cells was assessed by TUNEL,and the expressions of bax,bel-2,transforming growth factor-β2 (TGF-β2 ) in TCFs were detected using immunochemistry.Results The cultured cells showed the features of the fibroblasts in shape with the positive response for vimentin.A number of TUNEL positive cells were seen in Tet group and no TUNEL positive response was found in control group.The expression levels (A value) of bax,bcl-2 and TGF-~ protein in TCFs were 0.577 ± 0.009,0.430±0.012 and 0.341 ±0.017 in Tet group,and those in control group were 0.320±0.015,0.819±0.021 and 0.624±0.014 respectively,showing statistically significant differences between two groups( t =33.277,-35.356,-28.093,P＜0.01 ).Conclusions Tet suppresses the proliferation of human TCFs through up-regulating the expression of bax and down-regulating the expressions of bcl-2 and TGF-β2 in vitro.

The effect of microgravity on tissue structure and function of rat testis

To explore whether an environment of weightlessness will cause damage to the reproductive system of animals, we used the tail-suspension model to simulate microgravity, and investigated the effect of microgravity on the tissue structure and function of the testis in sexually mature male rats. Forty-eight male Wistar rats weighing 200-250 g were randomly assigned to three groups (N = 16 each): control, tail traction, and tail suspension. After the rats were suspended for 7 or 14 days, morphological changes of testis were evaluated by histological and electron microscopic methods. The expression of HSP70, bax/bcl-2 and AR (androgen receptor) in testis was measured by immunohistochemistry. Obvious pathological lesions were present in the testis after the rats were suspended for 7 or 14 days. We detected overexpression of HSP70 and an increase of apoptotic cells, which may have contributed to the injury to the testis. The expression of AR, as an effector molecule in the testis, was significantly decreased in the suspended groups compared to control (P < 0.01). We also observed that, with a longer time of suspension, the aforementioned pathological damage became more serious and some pathological injury to the testis was irreversible. The results demonstrated that a short- or medium-term microgravity environment could lead to severe irreversible damage to the structure of rat testis.