Anticonvulsant and Sedative Effects of Eudesmin isolated from Acorus tatarinowii on mice and rats.

This paper was designed to investigate anticonvulsant and sedative effects of eudesmin isolated from Acorus tatarinowii. The eudesmin (5, 10, and 20 mg/kg) was administered intraperitoneally (i.p.). The maximal electroshock test (MES) and pentylenetertrazole (PTZ)-induced seizures in male mice were used to evaluate anticonvulsant activities of eudesmin, and sedative effects of eudesmin were evaluated by pentobarbital sodium-induced sleeping time (PST) and locomotor activity in mice. Finally, the mechanisms of eudesmin were investigated by determining contents of glutamic acid (Glu) and gamma-aminobutyric acid (GABA) in epileptic mice, and expressions of glutamate decarboxylase 65 (GAD65), GABAA , Bcl-2, and caspase-3 in the brain of chronic epileptic rats. Results of MES and PTZ tests revealed that eudesmin possesses significant anticonvulsant effects, and the PST and locomotor activity tests demonstrated that eudesmin has significant sedative effects. Furthermore, our study revealed that after treatment with eudesmin, GABA contents increased, whereas Glu contents decreased, and ratio of Glu/GABA decreased. Our results also indicated that expressions of GAD65, GABAA, and Bcl-2 were up-regulated by treating with eudesmin, whereas the caspase-3 obviously was down-regulated. In conclusion, eudesmin has significant anticonvulsant and sedative effects, and the mechanism of eudesmin may be related to up-regulation of GABAA and GAD65 expressions, and anti-apoptosis of neuron the in brain.

[Correlation between the expression of perforin in T-lymphocyte subsets and antituberculosis in mice vaccinated with bacillus calmette-guerin].

OBJECTIVE: To investigate the relationship between the expression of perforin in T-lymphocyte subsets and antituberculosis in mice vaccinated with bacillus calmette-guerin (BCG). METHODS: 120 KM male mice were divided into a Control group (group C, n = 40, 20 each in group C1 or C2), a BCG vaccinated group (group B, n = 40, 20 each in group B1 or B2), a Tuberculosis group (TB group, n = 20), and a BT group, in which the mice were attacked by Mycobacterium tuberculosis H(37)Rv (MTB) after acquired immunity by vaccination with BCG (n = 20). Initially, mice in group B1, B2 and BT were vaccinated with BCG synchronously. Three month later, mice in group BT and TB were attacked by MTB synchronously. Samples of blood, lung, liver and spleen of mice in group C1 and B1 were collected at the same time. One month later again, Samples of blood, lung, liver and spleen of mice in group C2, B2, BT and TB were collected at the same time. Samples of blood were assayed for T-lymphocyte subsets expressing perforin (PFN(+)). The positive numbers of PFN(+) [CD(3)(+), CD(4)(+), CD(8)(+), CD(4)(+)CD(8)(+) double positive (CD(4)(+)CD(8)(+))] T lymphocytes and their percentages were assayed by flow cytometer. Specimens of lung, liver and spleen were examined for pathology and bacteriology. RESULTS: All the mice in the TB group acquired tuberculosis and the mortality was 55% (11/20) within 1 month. There were no tuberculosis and no death in mice of C, B and BT groups during the observation period. The amount of PFN(+) CD(8)(+) T lymphocytes in B group [(5.9 ± 0.9) × 10(3)] was significantly higher than that in C group [(4.8 ± 0.8) × 10(3)] (F = 42.24, P < 0.01). The PFN(+) CD(8)(+)% in TB group [(5.6 ± 0.9)%] was significantly less than that in B group [(7.3 ± 1.1)%] (F = 35.51, P < 0.05). For Mice in the BT group, the amount of PFN(+) (CD(3)(+), CD(8)(+), CD(4)(+)CD(8)(+)) T lymphocytes [(20.1 ± 5.5) × 10(3), (8.7 ± 0.4) × 10(3), 72 ± 19] and their CD(3)(+)%, CD(8)(+)%, CD(4)(+)CD(8)(+)% [(23.3 ± 3.3)%, (10.7 ± 1.6)%, (0.084 ± 0.015)%] were all higher than those in the B group [(13.0 ± 3.2) × 10(3), (5.9 ± 0.9) × 10(3), 36 ± 5, (15.5 ± 1.7)%, (7.3 ± 1.1)%, (0.044 ± 0.007)%] or the C group [(11.1 ± 3.0) × 10(3), (4.8 ± 0.8) × 10(3), 30 ± 7, (14.9 ± 1.7)%, (6.7 ± 0.9)%, (0.040 ± 0.006)%] or the TB group [(12.6 ± 1.6) × 10(3), (5.0 ± 0.1) × 10(3), 31 ± 3, (14.0 ± 1.7)%, (5.6 ± 0.9)%, (0.035 ± 0.005)%] (F = 14.23 - 74.98, P < 0.01 or P < 0.05), and the CD(4)(+)/CD(8)(+) in BT group (0.54 ± 0.17) was significantly lower than that in C group (0.76 ± 0.22) (F = 4.54, P < 0.01). CONCLUSIONS: Pre-vaccination with BCG increased PFN(+) CD(8)(+) T lymphocytes in the host. Acquired immunity by BCG vaccination can protect the host from attack by Mycobacterium tuberculosis. Increased PFN(+) (CD(3)(+), CD(8)(+), CD(4)(+)CD(8)(+)) T lymphocytes may be involved. The expression level of PFN by T lymphocytes may be an important marker for antituberculosis immunity.

[Analysis of T lymphocyte subsets and their expression of 4 cytokines in tuberculosis mice].

OBJECTIVE: To investigate the relationship of anti-tuberculosis immunity with perforin (PFN), granzyme B (GzmB), interferon-gamma (IFN-gamma), and interleukin-2 (IL-2) expression by T lymphocyte subsets. METHODS: Sixty mice were randomly allocated into a tuberculosis group and a control group (n = 30 each). Surface markers of T lymphocytes were stained with CD(3)PerCP, CD(4)FTTC, CD(8)APC, and intracellular cytotoxic molecules with PE-PFN, PE-GzmB, PE-IFN-gamma, and PE-IL-2 multi-color-labeled monoclonal antibodies, and analyzed at the single cell level. The relation between T lymphocyte subsets expression PFN, GzmB, IFN-gamma, IL-2 and antituberculosis immunity by flow cytometer. RESULTS: (1) CD(4)(+), CD(8)(+), and CD(4)(+)CD(8)(+) (DP) T lymphocytes all expressed PFN, GzmB, IFN-gamma and IL-2 to some degrees. The expressions of PFN and GzmB were much higher in CD(8)(+) T lymphocytes than those in CD(4)(+) T lymphocytes, while the expressions of IFN-gamma and IL-2 were higher in CD(4)(+) T lymphocytes. (2) The counts of T lymphocyte subsets and the percentages of T lymphocyte subsets to total lymphocytes may or may not reflect the cellular immunity consistently. (3) There was no significant difference in T lymphocytes expressing PFN between the tuberculosis group and the control group. But the counts of CD(3)(+), CD(4)(+), DP and CD(8)(+) T lymphocytes and the percentages of CD(3)(+), DP and CD(8)(+) cells expressing GzmB were significantly increased in the tuberculosis group (t value from -3.72 to 4.13, all P < 0.05). (4) IFN-gamma expressing CD(3)(+) and CD(4)(+) lymphocytes were increased significantly in the tuberculosis group. The counts of CD(8)(+) and DP T lymphocytes and the percentages of CD(3)(+), CD(4)(+), CD(8)(+), and DP cells that expressed IL-2 were decreased significantly in the tuberculosis group (t value from 2.62 to 3.46, all P < 0.05). CONCLUSION: CD(4)(+), CD(8)(+) and DP lymphocytes all can express PFN, GzmB, IFN-gamma and IL-2 at different degree levels.

[Immunotherapeutic efficacy of BCG vaccine in pulmonary tuberculosis and its preventive effect on multidrug-resistant tuberculosis].

OBJECTIVE: To evaluate the effect and safety of BCG vaccine on initially treated pulmonary tuberculosis and its controlling effect on multidrug-resistant tuberculosis. METHODS: All 360 volunteers with initially treated pulmonary tuberculosis of positive smear and culture were divided into immunotherapy group (180 cases, also BCG group) and control group (180 cases) at random pair. The patients in BCG group were treated with chemotherapy of a regimen of 2HRZ/2HR and immunotherapy with BCG for 4 months,and the first BCG vaccine was given a month after chemotherapy. Meanwhile, the patients in the control group were treated with chemotherapy of 2HRZ/4HR only. RESULTS: (1) The negative conversion rate of sputum smear in BCG group was 98.3% (177/180), and it was 97.2% (175/180) in control group. There was no significant difference between the two groups both at the ends of 4 and 6 months after treatment (chi2 = 0.1278, P > 0.05). (2) The positive conversion rate of sputum smear in BCG group was 2.3% (4/177), and it was 6.9% (12/175) in control group followed up for 5 years. The successful rate was 96.1% (173/180) in BCG group, and it was significantly higher than that of 90.6% (163/180) in control group (chi2 = 4.4643, P < 0.05). (3) In the 5-year follow up, bacteriologic result was similar to that of X-ray. (4) The occurrence rate of multidrug-resistant tuberculosis was 2.3% (4/177) in BCG group,significantly lower than that of 7.3% (13/178) in the control group (chi2 = 4.9513, P < 0.05). CONCLUSION: As an adjunct chemotherapy,immunotherapy with BCG vaccine should be helpful for patients with initially treated pulmonary tuberculosis. It would further strengthen the effects of chemotherapy and reduce the occurrence rate of multidrug-resistant tuberculosis.