The purposes of this study were to set up the method of the natural killer(NK) cell activity assay using the flow cytometer and to examine the characteristics and distribution of the NK cell during rat hepatocarcinogenesis. Forty five male 6 week-old specific pathogen free(SPF) Sprague-Dawley rats were randomly divided into three groups. Group I was the non-treated control and given normal diet and water. Group II was treated with diethylnitrosamine(DEN, 200mg/kg, i.p.) and partial hepatectomy. Group III was treated with DEN, partial hepatectomy and 0.05% phenobarbital sodium in water from 3 to 16 weeks. All animals were examined the morphology of the large granular lymphocyte(LGL), the LGL percent of the total lymphocytes and the LGL conjugation rate with YAC-1 cell in peripheral blood, spleen and liver. Moreover, activity of the LGL isolated from peripheral blood lymphocytes was determined using the flow cytometer. As results, LGL were observed in the peripheral blood, spleen and liver. LGL were observed the relatively faintly staining basophilic cytoplasm with granules, and eccentric, often kidney-shaped nuclei in Giemsa stain. Its size was $11{\sim}13{\mu}m$. LGL percentage of the isolated lymphocytes in peripheral blood, spleen and liver were 1.8~2.3%, 1.3~1.4% and 0.87~0.99%, respectively. LGL conjugation rate with YAC-1 cell was shown to be peripheral blood(9.3~10.3 %) > spleen(7.7~8.7%) > liver(5.6~7.0%). The activity of the LGL isolated from peripheral blood lymphocytes in Group I, II and III was 33.7%, 30.5% and 35.4%, respectively. However, all values were not significantly between groups.
These experiments were carried out to examine the development capacity of sexed and then bisected embryo from 8-cell to morula stage. Antisera to histocompatibility-Y(H-Y) antigen were prepared in inbred SD female rat by repeated immunization of spleen cell or testis supernatant from males of same strain. Male and female embryos were separated by delaying development of embryos against H-Y antibody. After sexing, rabbit embryos were bisected and aggregated. The results obtained from the these experiments were summuerized as follows: 1. When mouse and rabbit 8-, 16-cell and morular embryos were culature in H-Y antiserum, the ratio of embryo which has developed to hatching blastocyst was 53.4, 46.3 and 57.4% in mouse embryos, and 49.0, 52.0 and 61.0% in rabbit embryo, respectively. The ratio of mouse and rabbit embryos developed to hatching blastocyst showed nearly natural sex rate(50%), except rabbit mourla showed a little higher ratio(61.0%) as compared to natural sex ratio. 2. When rabbit demi-embryos from 8-, 16-cell embryo and morula were cultured, the percentage of demi-embryos was 70, 68 and 58% without zona pellucida removed, and 62, 69 and 51% with zona pellucida. The rate of aggregation was higher in 8- and 16-cell demi-embryos than in morula demi-embryo. 4. When sexed-demi-embryo was aggregated with another demi-embryo with demi-embryo with same sex, the rate of embryo developed to blastocyst was 60, 50 and 25%, respectively. Eight-cell demi-embryo showed highest rate. In conclusion, it showed that H-Y antiserum which was made by rat spleen cell enabled sexing rabbit embryos. And when rabbit sexed 8-, 16-cell and morula demi-embryo were aggregated, they were developed to eu-blastocyst which suggested the potential of sexed embryo manipulation.
Objective: The purpose of this study was to investigate the effect of Hwanggigunjungtang(HGT) on white rats with deteriorated immunity caused by methotrexate. Methods: First, methotrexate was fed to the white rats once a day for 4 days. After the immune responses of the rats are deteriorated, dried extracts of HGT mixed in water was fed to the rats once a day for 14 days. And then we measured the percentage of B-cell and T-cell in peripheral blood, the percentage of CD3+CD4+ T-cell and CD3+CD8+ T-cell of blood sampled from spleen and peripheral region. Results: The percentage of B-cell of peripheral blood was not different statistically. The percentage of T-cell of peripheral blood was increased significantly in HGT group as compared with control group. The percentage of CD3+CD4+ T-cell of peripheral blood was increased significantly in HGT group as compared with control group. The percentage of CD3+CD8+ T-cell of peripheral blood was decreased in HGT group as compared with control group. The percentage of CD3+CD4+ T-cell of spleen was increased significantly in HGT group as compared with control group. The percentage of CD3+CD8+ T-cell of spleen was not different statistically. Conclusion: According to the above results, HGT has an effect of increasing immune responses on white rats with deteriorated immunity caused by methotrexate.
Experiments were performed on rats to investigate the effect of doses of capsaicin on the immune response. Olive oil and the 0.3 mg, 1.0 mg and 3.0 mg/kg administration of capsaicin in olive oil were injected intraperitoneally every day for 4 weeks. Rats were sensitized and challenged with sheep red blood cells (S-RBC). Immune responses were evaluated by organ weight, HA and HY titer, Arthus reaction, delayed type hypersensitivity (DTH) and Rosette froming cell. Following results have observed. 1) The weight of spleen and thymus were increased by doses of compared with control group, but the body weight of rats was decreased. 2) HA titer, Arthus reaction and DTH were significntly decreased by doses of capsaicin as compared with control group. 3) Rosette forming cell in spleen cells was decreased according to the increase of capsaicin doses. These results suggested that high dose of capsaicin decrease humoral and cellular immune response in rats.
Cytochrome P-450(CP-450) is one of the three components of the liver microsomal enzyme system which hydroxylates fatty acids, hydrocarbons and a variety of drugs and other foreign compounds. Female Balb/c mice were immunized with purified polychlorinated bipheny(PCB)-induced CP-450 LMII. The spleen cells derived from immunized mice were fused with $SP^2$ myeloma cells using polyethylene glycol(PEG 3500). The hybrid cells were selected by hypoxanthine-aminopterine and thymidine(HAT) medium and the culture fluid were screened by enzyme-linked immunosorbent assay to CP450 LMII. The hybrid cess(${\times}10^7$) were innoculated into intraperitoneal cavity of Balb/c mice for the purpose of production of ascitic fluids. Monoclonal antibody(Mab) was purified from ascitic fluid by DEAE cellulose ion exchange chromatography and $I^{125}$-labeled Mab was also confirmed by autoradiography and SDS-polyacrylamide gel electrophoresis (MW : 55,000 and 110,000).
This study was designed to investigate the distributions of the positive cells in rat spleens by two monoclonal antibodies of MT1 and MB2. The spleens of immature 10 rats (Sprague Duwely, approximately 200gm) were collected and paraffin-embedded sections of spleens were stained with immunohistochemical methods. Higher proportions of MT1-positive cell number in spleens were ordered as marginal zone(8.5~18.1%), red pulp(2.1~8.8%) and periarterial lymphoid sheath(0~1.6%) in white pulp, and those of MB2-positive cell number are ordered as the central area of the periarterial lymphoid sheath(100%), red pulp(29.1~45.0%), marginal zone(15.2~30.4%), and peripheral area of periarterial lymphoid sheath(2.3~3.5%). The positive cells by MB2 are more numerous in number than by MT1. The above results were concluded that the positive cells by above two monoclonal antibodies were scattered throughout the red pulp and marginal zone, but in the central area of the periarterial lymphoid sheath, the MB2-positive cells only were present.
Sprague Dawley rats were exposed to electric fields (6,000 V, 10 kV/m, 30 min/day, 6 days per week) and injected strontium 90 (681 kbq/rat, one time) through abdominal cavity (strontium 90 and electric field complexed exposure group). In parallel, series with the electric field exposure only, strontium 90 injection only and control groups were run. Every group was consisted of 110 rats (55 male and 55 female). This animal experiment was performed from May to December in 1993. This results were conducted to investigate the effect of electric field for 11 weeks. The results are summarized as follows: 1. Ornithine decarboxylase (ODC) activity in rat's bone marrow cells: The ODC values was significantly increased in Sr$^{90}$ injection group and Sr$^{90}$ and electric field complexed exposure group as compared with that of control group (p<0.05). The ODC value was significantly decreased in electric field and Sr$^{90}$ complexed exposure group in comparison with Sr$^{90}$ injection group (p<0.05). The ODC values of electric field only exposure group was not different to that of control group (p>0.05). 2. The amount of Sr$^{90}$ accumulation in the femur, kidney and spleen:The accumulation amount of Sr$^{90}$ in the femur of Sr$^{90}$ injection group represented higher value than that of electric field and Sr$^{90}$ complexed exposure group (p<0.05). In the kidney and spleen, the difference between electric field and Sr$^{90}$ complexed exposure group and Sr$^{90}$ injection group wasn't observed. 3. The counts of white cells in blood of Sr9?injection group was decreased as compared with the value from control group and electric field and Sr$^{90}$ complexed exposure group (p<0.05). The rat's body weight, red blood cell counts and the weight data of liver, kidney and spleen did not show differences among four groups.
Kim, Jang-Oh;Choi, Jun-Hyeok;Shin, Ji-Hye;Jung, Do-Young;Min, Byung-In
Journal of radiological science and technology
/
v.39
no.3
/
pp.451-459
/
2016
The development of radioprotector is being actively conducted in order to reduce the damage from over radiation exposure at radiation accident or radiation therapy. So this study was confirmed for radiation protective effects using the Cynanchi wilfordii Radix that has been known to be effective for antioxidant activity, anti-cancer, immune enhancing effects. The method of this study was administered orally Cynanchi wilfordii Radix ethanol extracts to Sprague Dawley Rat(SD Rat) for 14 days once a day, while measuring changed blood cell, spleen index, liver and uterus tissue along the change in time of 1, 4, 7 and 21 days after X-ray beam of 7 Gy irradiation. As the result of the experiment, the experimental group's rats which are administered with Cynanchi Wilfordii Radix ethanol extracts showed a rapid recovery in white blood cell count(p < 0.05) and spleen index(p < 0.05). In addition, condensation of nuclei, cytoplasmic swelling, and inflammatory cell infiltration in experimental group's liver cell was decreased more than in irradiation group's component. Further, experimental group's Uterine gland decreased the apoptosis more than irradiation group's components did. It is expected that Cynanchi Wilfordii Radix extracts will be useful as a new radioprotector. With above in mind, this paper may provide appropriate implications with the field of emergency management such as radiation accident.
Abel et al. in Germany discovered a new dioxin-responsive gene, which has later been identified as rpt-1 (regulatory protein T-lymphocyte 1). While it is speculated that rpt-1 may play a role in signal transduction and carcinogenesis, its roles and functions remain unknown. The present study attempted to analyze functions of rpt-1 in human epithelial cells following the xenobiotic exposures. While German counterpart analyzed expressionn of rpt-1 in spleen and thymus cells from mouse and rat and characterizes molecular properties of the gene, our work mainly focused on analyzing function of rpt-1 in human skin cells. Expression of rpt-1 in human cells were analyzed by western and northern blot RT-PCR analysis. Expression of rpt-1 as well as Staf-50 in human cells with or without exposure to environmental pollutants were also analyzed by northern blot analysis, since Staf-50 is homologous with rpt-1 and found in human cells. To help study roles of rpt-1 in human cell system, retroviral vector system carrying rpt-1 gene under the CMV promoter were constructed and transfected. Cells overexpressing the gene after the transfection showed an increase of cell density and soft agar colony formations, as compared to the control cells, suggesting that rpt-1 may play a certain role in the transformation processes of human cells. While the expression of rpt-1 in spleen and thymus is known to be strong in the laboratory animals, both the basal and TCDD-induced expression of rpt-1 in the current cellular system remained insignificant. It is speculated that the expression pattern of rpt-1 may be tissue- and species-specific. The present study demonstrated a strong expression of rpt-1 protein in the brain of SD rat model. Since there is no previous report on the expression of rpt-1 in the brain tissue, the result may play a significant role in understanding dioxin-induced neurotoxicities in the future. The present study provides an opportunity to understand a role of rpt-1 in human cell system and suggest a possible lead and basis for the future study of dioxin-induced neurotoxicities.
Objectives : The purpose of this study is to investigate the effect of oral administration of Yongyukjowi-tang(YJT) on the immune activity in aged Sprague-Dawley rat(SD rat). Methods : SD rats were divided into three groups: Yongyukjowi-tang groups(YJT groups), distilled water groups(control groups) and Vitamin C groups(positive control groups) which were administered an oral dose(1ml/1day) to 6, 48 and 68 weeks old SD rats for four weeks. After four weeks, the number of total leukocyte, CD3+, CD4+, CD8+, and the level of cytokine (IL-2, IL-4, IL-10, IFN-${\gamma}$) were measured in spleen tissue of each SD rats. Results and Conclusions : 1) The number of total leukocyte significantly increased in 52 weeks old YJT group and 72 weeks old YJT group in comparison with those of the control group. 2) The number of CD3+ cell significantly increased in 72 weeks old YJT group in comparison with those of the control group and the positive control group. 3) The number of CD8+ cell significantly increased in 52 weeks old YJT group in comparison with those of the control group. 4) The level of IL-2 significantly increased in 72 weeks old YJT group in comparison with those of the control group and the positive control group. 5) The level of IL-4 significantly increased in 52 weeks old YJT group in comparison with those of the positive control group. These results suggest that oral administration of YJT has an effect on increase of immune activity in aged rat.
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