ABSTRACT
Objective To determine the difference of macrophage RAW264.7 apoptosis induced by Brucella melitensis virulent strain 16M and attenuated strain M5-90 and elucidate the regulatory role of caspases 3,8 and 9.Methods The best multiplicity of infection (MOI) was determined through kinetic analysis of Brucella melitensis strain 16M and M5-90 induced mouse macrophages apoptosis(bacterium ∶ cell =100 ∶ 1,50 ∶ 1,10 ∶1).The infection model was established using the best MOI =50 ∶ 1.The numbers of in vivo bacteria by colony formation units were calculated after macrophages were infected for different times,including 2,4,8,12,24 and 48 h,and the infected cells were collected.The ratios of apoptosis were detected and the regulation of caspases 3,8 and 9 in apoptosis pathway was elucidated by flow cytometry.Results The numbers of 16M in vivo bacteria were 105.4,104.8,105.8,106.5,108.0 and 109.0,respectively and of M5-90 were 106.1,106.2,106.4,106.3,106.1 and 105.0,respectively.The number of in vivo bacteria of 16M was significantly increased than that of M5-90 after infected for 24 h to 48 h.The ratios of apoptosis induced by 16M after infected for 2,4,8,12,24 and 48 h was (2.67 ± 0.09)％,(13.13 ± 0.30)％,(6.56 ± 0.42)％,(6.49 ± 0.28)％,(16.07 ± 0.86)％ and (24.23 ± 1.67)％,respectively,and by M5-90 was (3.62 ± 0.02)％,(32.01 ± 2.59)％,(17.58 ± 0.44)％,(16.09 ± 0.10)％,(62.53 ± 2.70)％ and (85.53 ± 0.15)％,respectively,and by control group was [(1.90 ± 0.20)％,(1.92 ±0.16)％,(1.99 ± 0.03)％,(2.48 ± 0.11)％,(3.56 ± 0.07)％,(5.26 ± 0.33)％].The differences were statistically between groups in same time.The Brucella melitensis vaccine strain M5-90 was more powerful than virulent strain 16M in respect of inducing macrophage apoptosis after infected for 24 to 48 h.Twenty-four hours after infection,the expression of caspases 3,8 and 9 was (1.47 ± 0.05)％,(1.52 ± 0.02)％ and (2.47 ± 0.12)％,respectively,in control group and the expression was (9.70 ± 0.46)％,(6.08 ± 0.56)％ and (35.08 ± 1.64)％,respectively,after infected for 24 h induced by M5-90.The expression of caspases 3,8 and 9 was significantly higher than that control group (P ＜ 0.01).Twenty-four hours after given caspases 3,8 and 9 inhibitor,apoptosis rate in control group was (66.72 ± 1.28)％,in M5-90 group was (22.58 ± 0.55)％,(53.15 ± 1.85)％ and (29.18 ± 0.23)％,respectively,and compared with control group,apoptosis rate of caspases 3,8 and 9 was significantly lower(P ＜ 0.01).Conclusions Apoptosis of macrophage can be induced by Brucella melitensis virulent vaccine strain 16M and attenuated strain M5-90.M5-90 is stronger than that of strain 16M.Caspases 3,8 and 9 can regulate macrophage apoptosis after M5-90 infection.
ABSTRACT
<p><b>OBJECTIVE</b>To observe the effects of training on plasma content of NO, ET and NO/ET system in rats fed with high methionine diets.</p><p><b>METHODS</b>Male Wistar rats were randomly divided into normal diet group (CR), high methionine diet group (MR) and high methionine diet plus 90 min swimming group (T+ MR). Rats in MR and T+ MR group were fed with the diet which contained 3%of methionine for eight weeks, but rats in control group were fed with ordinary diet. After breeding for eight weeks homocysteine (Hcy) level, T-NOS, NO and ET were measured.</p><p><b>RESULTS</b>Plasma homocysteine of MR increased twofold as compared with that of CR,whereas plasma NO/ET of MR descended significantly (P < 0.05), suggesting that hyperhomocysteinemia was induced by the high methionine diet, and plasma NO/ET system was out of balance. Plasma homocysteine content of T+ MR obviously descended, plasma T-NOS, NO and NO/ET ascended obviously (P < 0.05) as compared with that of MR, and the above indicators of group T + MR were not different as compared with that of CR (P > 0.05).</p><p><b>CONCLUSION</b>Proper training can descend plasma homocysteine level of rats fed with high methionine diet and better balance between NO and ET to prevent developing of hyperhomocysteinemia.</p>
Subject(s)
Animals , Male , Rats , Dietary Supplements , Endothelins , Blood , Hyperhomocysteinemia , Methionine , Nitric Oxide , Blood , Nitric Oxide Synthase , Blood , Physical Conditioning, Animal , Physiology , Rats, Wistar , Swimming , PhysiologyABSTRACT
<p><b>OBJECTIVES</b>To investigate the pharmacological effects of azidothymidine (AZT) on p33ING1b expression, senescence and apoptosis of TJ905 glioblastoma cells.</p><p><b>METHODS</b>TJ905 cells were treated with AZT at a serial concentrations of 50, 100 and 200 µmol/L. Semi-quantitative RT-PCR and cytochemical staining of senescence related-galactosidase (sβ-Gal) were used to evaluate the expression of p33ING1b mRNA and to label the senescent cells at the 1st, 3rd and 6th generations, respectively. In situ cell death detection and single cell gel electrophoresis were used to detect the apoptosis at the 3rd and 6th generations.</p><p><b>RESULTS</b>AZT induced the expression of p33ING1b mRNA and senescence of the tumor cells of the 1st generation in a dosage and time dependent manner. At the 6th generation, the relative amount of p33ING1b RT-PCR product (1.44±0.23) and sβ-Gal labeling index of 200 µmol/L group (45.62±6.74) were significantly higher than those of the 1st (0.95±0.13 and 7.82±2.40) and the 3rd generation cells (1.35±0.23, 26.27±7.17) of the same group, and cells of the same generation in the 50 µmol/L (0.85±0.24, 27.37±6.41) and 100 µmol/L groups (1.23±0.34, 35.49±5.12, P<0.01). There was a significant positive correlation between the p33ING1b mRNA expression and the labeling index of sβ-Gal. Pro-apoptotic effects of AZT became obvious at the 6th generation.</p><p><b>CONCLUSION</b>AZT upregulates the expression of p33ING1b, a possible mechanism in regulating senescence and apoptosis of the TJ905 cells.</p>
Subject(s)
Humans , Apoptosis , Brain Neoplasms , Metabolism , Pathology , Cell Line, Tumor , Cellular Senescence , Dose-Response Relationship, Drug , Glioblastoma , Metabolism , Pathology , Inhibitor of Growth Protein 1 , Intracellular Signaling Peptides and Proteins , Genetics , Metabolism , Nuclear Proteins , Genetics , Metabolism , RNA, Messenger , Metabolism , Reverse Transcriptase Inhibitors , Pharmacology , Reverse Transcriptase Polymerase Chain Reaction , Tumor Suppressor Proteins , Genetics , Metabolism , Zidovudine , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of human beta-defensin-2 (HBD-2) mRNA in skin lesions of patients with recurrent genital herpes (RGH) and the effect of Huangbai Liquid (HL) on it.</p><p><b>METHODS</b>Twenty-seven patients were randomly assigned to 2 groups, the HL group (n = 14) treated with HL and the famciclovir group (n = 13) with famciclovir. HBD-2 expression of patients were detected before and after the treatment and compared with that of 10 healthy subjects.</p><p><b>RESULTS</b>HBD-2 expression was found in the skin lesions of both the healthy persons and the RGH patients before treatment. It is higher significantly in the HL group than in the famciclovir group after treatment.</p><p><b>CONCLUSION</b>HL was suitable for treatment of RGH since it could improve immune function of RGH patients and keep a rather higher concentration of HBD-2 expression in local skin lesions.</p>