ObjectiveTo study the influence of myxobacteria metabolites NX52 and NX83 on the proliferation of colorectal carcinoma cells and investigate its probable mechanism. MethodsThe human colorectal carcinoma cell lines HT-29, SW480, and SW1463 were respectively treated with the two metabolites (NX52 and NX83) at different concentrations (0.1 mg/mL, 1.0 mg/mL, and 10.0 mg/mL), the cells of negative control were treated without metabolite. The proliferation inhibition was examined by methyl tthiazolyl tetrazolium assay. The cell morphology character was compared by inverted microscope, and the apoptosis of cell was analyzed by flow cytometry. ResultsTwo kinds of metabolites NX52 and NX83 had time-dose inhibitory effects on proliferation of human colorectal carcinoma cells HT-29, SW480, and SW1463 (P < 0.05). The metabolite NX83 had more obvious proliferation inhibition in the colorectal carcinoma cells as compared with the metabolite NX52 (P < 0.05). After 48 h, the apoptosis rate of the metabolite NX83 for SW1463 cell was observably increased as compared with the negative control group (P < 0.01). ConclusionsThe two kinds of metabolites NX52 and NX83 from myxobacteria could kill colorectal carcinoma cells in vitro. The possible mechanism might be induced by apoptosis of tumor cells.
Objective To observe the expression of cyclin dependent kinase 5 (Cdk5) and p25 in the pathogenesis of retinitis pigmentosa (RP) in Royal College of Surgeon (RCS) rats and its relationships with apoptosis. To explore the mechanism of Cdk5 and p25 induced photoreceptor apoptosis in the pathogenesis of RP. Methods Retinas of RCS and RCS-rdy+ rats were obtained at the ages of postnatal day 17, 25, 35, 60. The retinal structure and thickness of outer nuclear layer were measured by optical microscopy. The expression of Cdk5, p25, cleave-caspase 3 in the retina was evaluated by immunohistochemistry. The protein expression of cleave-caspase 3 in the retina was determined by Western blot. The apoptosis of retinal cells was examined by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL). The mean absorbance value of apoptotic cells was analyzed by SPSS 17.0 software. Results The retinal thickness of the RCS rats was significantly reduced in comparison to the RCS-rdy+ rats as the postnatal days progressed, particularly in the layer of rods and cones and the outer nuclear layer. The expression level of Cdk5, p25, cleave-caspase 3 of RCS rats increased from postnatal 17 days to postnatal 35 days, while decreased on postnatal 60 days; but there was no obvious change of above indexes in RCS-rdy+ rats. The protein expression of cleave-caspase 3 in the RCS rats was significantly increased with progression of postnatal days to postnatal 35; but there was no obvious similar change in RCS-rdy+ rats. The results of TUNEL showed that the apoptotic cells significantly increased in the outer nuclear layer of RCS rats from postnatal 17 days to postnatal 35 days, while decreased on postnatal 60 days; but there was no obvious change of above index in RCS-rdy+ rats. This study showed that there were significant correlations between the following variables: Cdk5 expression and p25 expression, Cdk5 expression and cleave-caspase 3 expression, Cdk5 expression and apoptotic cells, p25 expression and cleave-caspase 3 expression, p25 expression and apoptotic cells, cleave-caspase 3 expression and apoptotic cells. The partial correlation coefficients were 0.949, 0.808, 0.959, 0.887, 0.979, 0.852, respectively and the P value was 0.000. Conclusions The apoptotic cells significantly increases and the expression level of Cdk5, p25, cleave-caspase 3 of RCS rats increases from postnatal 17 days to postnatal 35 days. The tendency of apoptotic cells to increase is consistent with the change of Cdk5, p25, cleave-caspase 3 expression. The apoptosis of photoreceptor cells is related to increasing expression of Cdk5 and p25 in RCS rats. Cdk5 may be involved in the development of RP in RCS rats.
PURPOSES:To investigate the time of neuronie apoptosis in the retinas of Imman fetuses,and its relations with neuronie proliferation and differentiation, METHODS:The retinas of 27 human fetuses from 8th to 38th week of R,~til- ization age and 3 adults were studied by TdT-mediated dUTP nick end labelling(TUNEL) method. RESULTS:Tbe nuctei of labeled apoptotic cells were charaeterised by nuclear marginization,ehromatln condensation and cleseent shape,and some apoptotie bodies were visible in the specimens. The apoptosis of neuroepithelium of fetal rclina took place during 8th to 18th week, Apoptosis of ganglion cells were observed from 1256 to 18th week. The apoptos[s of pholorec, plors were formd from 14th to 2Ist week ,while thai of bipolar neurones and M~ller cells were found from ldth to 28th week. No apoptosb of ocstones were observed in the retinas after 28th week of fertilization age and within the retinas of adults. CONCLUSION:The proliferating cells of neuroepithelium and Ihe neurones which just differetiated from fetal retina might partly undergo apoptosis. The time of apoptosls of differentiated neurones was consistent with the time of the synapses formation between neurones and their targel cells. （Chin J Ocul Fundus Dis,1997,13:67 -69 ）
ObjectiveTo investigate the possible mechanism of cucurmosin on apoptosis in human pancreatic cancer cell line SW1990 in vitro. MethodsThe inhibition of cucurmosin on SW1990 cell was detected by MTT assay, the apoptosis was observed by transmission electron microscope, the apoptosis rate was analyzed by flow cytometry, and the protein level of caspase3 was determined by Western blot. ResultsAfter exposure to cucurmosin at 1.25, 2.50, 5.00, 10.00, 20.00, 40.00, and 80.00 μg/ml for 24, 48, and 72 h, the proliferation of SW1990 cell was inhibited in a time-and dose-dependent manner (Plt;0.05). At 72 h after 40.00 μg/ml cucurmosin treatment, the typical apoptosis changes and apoptotic bodies were observed by transmission electron microscope. After exposure to cucurmosin at 0, 2.50, 10.00, and 40.00 μg/ml for 72 h, the apoptosis rate increased gradually as (0.30±0.11)%, (18.93±1.06)%, (28.00±2.07)%, and (49.93±3.25)%, respectively (Plt;0.05). The expression of caspase-3 protein was elevated gradually (Plt;0.05). ConclusionCucurmosin may induce the apoptosis of pancreatic cancer cell through up-regulating the expression of caspase-3.
Objective To investigate an effect of compressive stress on proliferation and apoptosis of human hyperplastic scar fibroblasts（HSFb） in vitro. Methods HSFb were obtained from a 20 year old female patient who developed a hyperplastic scar 3 months after operation for a largearea burn. HSFb were isolated, and were cultured in vitro with the simplified airpressure controlled cellculture instrument, and then they were randomly divided into the following 8 groups: the control group (no stress) and the 7 continuous compressive stress groups, which respectively underwent the 5, 10, 15, 25, 50, 100 and 150mmHg(1mmHg=0.133 kPa) pressure treatment for 4d ays. The absorbance (A) of the cell and the inhibition ratio (IR) of the cell proliferation were determined by the MTT assay, the cell growth cycle was determined by the flow cytometer, and the cell apoptosis was observed by the AnnexinV binding with PI labeling method. Results In the 5, 10, 15, 25, 50, 100 and 150mmHg pressure groups and the control group, the A values of the cells were 0.228±0.004, 0.226±0.003, 0.213±0.005, 0.180±0.005, 0.172±0.007, 0.165±0.004, 0.164±0.004 and 0.230±0.005, respectively; the IRs of the cell proliferation were 0.8%，2.0%，7.3%，21.7%，252%， 28.2% and 0, respectively;the ratios of the cells in G1 were 71.80％±0.44％, 72.32％±0.40％, 74.56％±1.01％, 82.82％±2.76％, 86.77％±2.06％, 88.23％±1.27％, 89.11％±1.74％ and 71.6％±0.49％,respectively; the cell apoptosis ratios were 4.22％±0.49％, 5.12％±0.74％ , 8.58％±0.79%, 19.28％±1.40%, 25.60％±1.21%, 3580％±2.39%, 36.18％±2.38% and 4.00％±0.36％, respectively. In the 5 and 10mmHggroups there were no statistically significant differences in all the above parameters when compared with those in the control group (P＞0.05); however, in the 15, 25,50, 100 and 150mmHg groups there were statistically significant differences in the above parameters when compared with those in the control group (P＜0.05). Furthermore, in the 10, 15, 25 and 50 mmHg groups, there were statistically significant differences in the Avalue of the cells and the ratios of the cells in G 1 when compared with each other (P＜0.01). By contrast, there were no statistically significant differences in the 50, 100 and 150 mmHg groups when compared witheach other (P＞0.05). In the 10, 15, 25, 50 and 100mmHg groups there werestatistically significant differences in the cell apoptosis ratio when comparedwith each other (P＜0.01). In the 100 and 150 mmHg groups there were no such statistically significant differences when compared with each other (P＞0.05).Conclusion A continuous compressive stress when given properly can have a combined effect of the proliferation inhibition and the apoptosis promotion on HSFb in vitro, and this kind of combined effects can becomeone of the important mechanisms for the pressure therapy in treating hyperplastic scar.
【Abstract】Objective To research relation of apoptosis muscular cell in 103Pd radioactive stent of dog biliary muscular formation and inhibition of biliary ductal stricture. Methods Twelve dogs were randomly divided into two groups, which were general stent group and 103Pd radioactive stent group. General stent and 103Pd radioactive stent were respectively put into extrahepatic biliary tract of two groups. After 30 days all dogs were killed, and biliary tract were taken out. Apoptotic cells were detected by immunohistochemical methodsand agar electrophoresis, and nucleus browyellow was positive cell. Dog biliary duct cross-sections were stained by hematoxylin-erosin; area and perimeter of lumen,thickness of inner membrane and stenosis degree in bile duct were analysed by image analysis software of computer.Results The apoptotic biliary duct smooth muscle cell ［(87.9±7.96)/cm2］ was more significantly increased in the 103Pd radioactive stent group than in the general stent group ［(5.6±0.51)/cm2］, P＜0.05; and comparing with the general stent group, the 103Pd radioactive stent significantly reduced biliary muscular formation thickness. Conclusion The result shows that 103Pd radioactive stent can inhibit proliferation of biliary ductal smooth muscle cell.
OBJECTIVE: To explore the effect of Fas/Apo-1 and Bcl-2 gene expression on mechanism of scar formation. METHODS: Immunohistochemical method was applied to defect the expression of Fas and Bcl-2 protein in fibroblasts from 10 cases with normal skin, 10 cases with hypertrophic scar and 10 cases with keloid. RESULTS: The positive expression rate of Bcl-2 protein in keloid was 83.2%, significantly higher than that in hypertrophic scar (38.6%), (P lt; 0.01), and the positive expression rate in hypertrophic scar and keloid was higher than that in normal skin (6.78%), (P lt; 0.01). But the positive expression rate of Fas/Apo-1 protein was 78.4% in normal skin 80.4% in hypertrophic scar, 84.4% in keloid respectively, which showed no significant difference among them (P gt; 0.05). CONCLUSION: Bcl-2 gene but Fas gene may take part in the formation of pathologic scar.
OBJECTIVE: To investigate the effect of vasostomy on apoptosis in the male rat spermatogenic cells after vasoligation. METHODS: Model of vasoligation and vasostomy in male rat was established, and then terminal deoxynucleotidyl transferase-mediated dUTP nick labelling technique to detect the apoptosis of spermatogenic cells at 4, 8, 12, 16 weeks after vasostomy. RESULTS: The number of apoptotic cells in vasostomy group was significantly lower than that of vasoligation group since 8 weeks after vasostomy(P lt; 0.05). The number of apoptotic cells in 8 and 12 weeks after vasostomy were significantly higher than that in prevasoligation(P lt; 0.05). 16 weeks after vasostomy, the number of apoptotic cells restored to the level same as that in prevasoligation stage. CONCLUSION: Vasostomy can reverse the apoptosis of spermatogenic cells due to vasoligation.
Objective To find the relation between donor liver cold preservation-reperfusion injury and hepatocellular apoptosis during liver transplantation. Methods Four groups of rabbit livers, which had experienced cold storage for 0,3,6,9hr respectively, were observed during their cold preservation-reperfusion by using TdT-mediated dUTP-biotion nickend labeling and electron microscope.Results Apoptopic hepatic parenchymal cells were obviously observed in reperfusing livers subsequent to cold storage. Furthermore, the longer the cold storage duration, the greater the number of apoptotic cells. On the contrary, no or rare apoptotic hepatic parenchymal cells was observed in all the groups at the end of cold preservation. Conclusion It suggests that apoptosis of hepatic parenchymal cells is markedly involved in donor liver cold preservation-reperfusion injury.
Objective To investigate whether the agonist of delta opoid receptor D-Ala(2),D-Leu(5) enkephalin (DADLE) has the effect of decreasing myocardial injury during ischemia-reperfusion of adult rabbits’ myocardium，so that a new mehanism and way to myocardial protection could be found. Methods Langendorff model was used during the experiment. Thirty rabbits were divided into three groups randomly (each group 10 rabbits). Control group: St.Thomas Ⅱ cardioplegic solution was used; group 1: St.Thomas Ⅱ cardioplegic solution and DADLE (1mg/kg) were used; group 2: St.Thomas Ⅱ cardioplegic solution and naloxone(3mg/kg) were used to induce the hearts to arrest respectively. After arrest the hearts were reperfused respectively. Data of left ventricle development pressure(LVDP) was recorded before and after ischemia. Biochemical indicators of myocardium, lactate dehydrogenase(LDH) were detected before and after ischemia. Some myocardial tissues were used to explore the changes of the tissue of ultrastructure with electron microscope，when the experiment was over. Still some myocardial tissues were to be detected by flow cytometer to evaluate the apoptosis of the myocardium. Results The LDH and LVDP showed significant difference among three groups after ischemia(Plt;0.05); LVDP in group 1 was higher than those in group 2 and control group(69.8±5.8 mmHg vs. 23.4±3.9 mmHg; 69.8±5.8 mmHg vs. 37.9±4.7 mmHg; Plt;0.05), the LDH in group 1 was lower than those in group 2 and control group(1 272.6±59.1 U/L vs. 2 764.4±27.7 U/L, 1 272.6±59.1 U/L vs. 1 884.4±37.5 U/L; Plt;0.05). The apoptosis rate in group 1 was lower than those in group 2 and control group. As could be shown from the ultrastructure: mitochondria structure was nearly normal in group 1; mitochondria structure was injuried severely in group 2; there was a minor injury in control group. Conclusion Agonist of δ opoid receptor DADLE in cardioplegic solution could induce hibernation, which has myocardial protection effect during ischemia-reperfusion injury.