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소동물에서 복강내 장기의식실험을 위한 이상적인 마취약 투여경로에 대한 제안
김준예 ( Joon Ye Kim ),이정익 ( Jeong Ik Lee ),방우휘 ( Yu Hui Fang ),정진호 ( Jin Ho Jeong ),김유선 ( Yu Seun Kim ) 한국조직공학·재생의학회 2008 조직공학과 재생의학 Vol.5 No.4
Intraperitoneal(lP) administration of anesthesia is commonly used for laboratory animal experiment including islet isolation using abdominal organ. However, the direct effects of anesthetics towards organs of abdom?inal cavity such as pancreas have been neglected. To evaluate the direct effects of anesthetic agents to the pancreas when isolating pancreatic islets, we compared the islet function and recovery yield from the rats that were anesthe?tized using IP and intramuscular(IM) routes. Lewis was anesthetized using the combination of zoletil and xyla?zine(Z/X). These drugs were injected through 1M or IP route. To shot 1M drugs, we have invented a specially designed restrainer. Glucose challenging test was performed with each group of prepared islets within 30 minutes after isolation procedure. Yield of isolated islets by 1M route was significantly improved(1M: 1136±226 ea, IP: 511±154 ea). Function of fresh islet with 1M route was better kept than oflP, which was expressed by stimulation index. For rodent islet isolation, 1M route for anesthesia is simpler and safe way compared with IP route. It is sug?gested that 1M route is versatile in laboratory animal experiments.
김수진,김준예,김은지,이효정,조유리,김명수,김유선,김범석,허규하 연세대학교의과대학 2022 Yonsei medical journal Vol.63 No.2
Purpose: The immunomodulatory effects of thalidomide (TM) and dexamethasone (DX) on immune cells and their co-stimulatory,co-inhibitory molecules in vitro and in vivo have been previously reported. The current study investigated the effects of TMand the combinatorial treatment with DX on immune cells using a murine cardiac allograft transplantation model. Materials and Methods: Intraabdominal transplant of cardiac allografts from BALB/c (H-2d) donors to C57BL/6 (H-2b) recipientswas performed. After transplantation, mice were injected daily with TM or DX or a combination of both TM and DX (TM/DX) by intraperitonealroute until the time of graft loss. CD4+ T cell subsets and CD11c+ cells in the peripheral blood mononuclear cells andspleen were examined and quantified with flow cytometry. Serum IL-6 levels were measured by enzyme-linked immunosorbent assayon day 7. Results: The mean graft survivals were 6.86 days in the untreated group, and 10.0 days in the TM/DX group (p<0.001). The TM/DXtreatment affected the CD4+ T cell subsets without suppressing the total CD4+ T cell population. The CD4+FOXP3+/CD4+CD44hi Tcell ratio increased. Increase in cell counts and median fluorescence intensity on CD11c+CD85k+ with TM/DX were observed. Theinhibition of pro-inflammatory cytokine interleukin-6 was also observed. Conclusion: These outcomes suggest the immunomodulating effect of the TM/DX combinatorial treatment. In conclusion, TM/DX combination may be a promising immunomodulatory approach for preventing allograft rejection and improving graft survivalby inducing tolerance in transplantation.
Antitumor Effect of Low-Dose of Rapamycin in a Transgenic Mouse Model of Liver Cancer
이형순,김준예,노원상,김명수,김혜령,주동진 연세대학교의과대학 2022 Yonsei medical journal Vol.63 No.11
Purpose: We investigate whether low-dose rapamycin is effective in preventing hepatocellular carcinoma (HCC) growth and treating HCC after tumor development in transgenic mice. Materials and Methods: We established transgenic mice with HCC induced by activated HrasG12V and p53 suppression. Trans genic mice were randomly assigned to five experimental groups: negative control, positive control, tacrolimus only, rapamycin only, and tacrolimus plus rapamycin. The mice were further divided into two groups according to time to commencement of im munosuppressant treatment: de novo treatment and post-tumor development. Results: In the de novo treatment group, marked suppression of tumor growth was observed in the rapamycin only group. In the post-tumor development group, the rapamycin only group displayed no significant suppression of tumor growth, compared to the positive control group. In T lymphocyte subset analysis, the numbers of CD4+effector T cells and CD4+ regulatory T cells were significantly lower in the positive control, tacrolimus only, and tacrolimus plus rapamycin groups than the negative control group. Immunohistochemical analysis revealed significantly higher expression of phosphorylated-mTOR, 4E-BP1, and S6K1 in the posi tive control group than in the rapamycin only group. Conclusion: Low-dose rapamycin might be effective to prevent HCC growth, but may be ineffective as a treatment option after HCC development.
김은지,김준예,최훈영,이효정,이주한,김명수,김유선,허규하,김범석 연세대학교의과대학 2021 Yonsei medical journal Vol.62 No.2
Purpose: In organ transplantation, the need for immune modulation rather than immune suppression has been emphasized. Inthis study, we investigated whether combinatorial treatments of with thalidomide (TM) and dexamethasone (DX) might be newapproaches to induce systemic immunomodulation on T cells and other immune cells that regulate the expression of co-inhibitorymolecules. Materials and Methods: Naïve splenic T cells from C57BL/6 mice were sort-purified and cultured in vitro for CD4+ T cell proliferationand regulatory T cell (Treg) conversion in the presence of TM or/and DX. Expression of cytotoxic T-lymphocyte-associatedantigen-4 (CTLA-4) and programmed death-1 (PD-1) in proliferated and converted T cells was quantified by flow cytometry. Wealso quantified in vivo expression of CTLA-4 and PD-1 on splenic CD4+ T cells and other immune cells isolated from TM- or/andDX-treated mice. Mixed lymphocytes reactions (MLR) were performed to evaluate the capacity of immune cells in carrying outimmune responses. Results: CTLA-4 expressions in effector T cells in vivo and in Tregs in vivo/vitro significantly increased upon TM/DX combinatorialtreatment. Corresponding to increased CTLA-4 expression in T cells, the expression of ligand molecules for CTLA-4 significantlyincreased in splenic dendritic cells in TM/DX-treated groups. In addition, MLR results demonstrated that splenocytes isolatedfrom TM/DX-treated mice significantly suppressed the proliferation of T cells isolated from other strains. Conclusion: Based on these results, we suggest that TM/DX combinatorial treatments might be efficient immunomodulatorymethods for regulating T cell immunity.