• 한국동물생명공학회지
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• 제37권3호
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• pp.169-175
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• 2022

Bovine mammary epithelial (MAC-T) cells are commonly used to study mammary gland development and mastitis. Lipopolysaccharide is a major bacterial cell membrane component that can induce inflammation. Autophagy is an important regulatory mechanism participating in the elimination of invading pathogens. In this study, we evaluated the mechanism underlying bacterial mastitis and mammary cell death following lipopolysaccharide treatment. After 24 h of 50 ㎍/mL lipopolysaccharide treatment, a significant decrease in the proliferation rate of MAC-T cells was observed. However, no changes were observed upon treatment of MAC-T cells with 10 ㎍/mL of lipopolysaccharide for up to 48 h. Thus, upon lipopolysaccharide treatment, MAC-T cells exhibit dose-dependent effects of growth inhibition at 10 ㎍/mL and death at 50 ㎍/mL. Treatment of MAC-T cells with 50 ㎍/mL lipopolysaccharide also induced the expression of autophagy-related genes ATG3, ATG5, ATG10, ATG12, MAP1LC3B, GABARAP-L2, and BECN1. The autophagy-related LC3A/B protein was also expressed in a dose-dependent manner upon lipopolysaccharide treatment. Based on these results, we suggest that a high dose of bacterial infection induces mammary epithelial cell death related to autophagy signals.

• 융합정보논문지
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• 제11권2호
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• pp.201-210
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• 2021

본 연구는 메밀 추출물과 비텍신을 이용하여 UVB 손상 개선에 대한 효과를 알아보기 위해 Microarray 분석, 세포의 증식, 세포 상처 회복, 세포주기, 마이크로파지의 생성 양상, 단백질 분석 등을 실시하였다. Microarray 분석 결과는 DRAM1, Atg2a 및 Atg13 유전자에서 UVB에 의해 증가 된 양상을 메밀 에탄올추출물과 비텍신에서 감소시켰다. 세포의 증식, 상처 회복, 주기 및 마이크로파지 양상에서는 메밀 에탄올추출물과 비텍신에서 정상 세포와 유사하게 개선되었으며, 단백질 분석에서 DRAM1, Beclin-1 및 LC3 I/II 모두 비텍신 처리군에서 감소하였고, p-mTOR 및 Survivin은 모두 증가 되었다. UVB에 의한 손상에서 메밀 에탄올추출물과 비텍신은 DRAM1, Atg2a 및 Atg13을 같이 컨트롤 하고 세포 증식, 상처 회복 및 주기를 정상으로 회복하며 UVB에 의한 세포 노화 발생원인 중 하나인 오토파지를 조절하여 세포의 사멸억제 및 재생하므로 기능성 화장품 성분으로 활용가능할 것으로 사료 된다.

• 대한물리의학회지
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• 제16권4호
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• pp.23-31
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• 2021

PURPOSE: This study compared the effect of training ankle joint and hip joint thera-band exercise on balance. METHODS: The participants were divided into two groups of 11 each. Group A performed hip exercise after ankle exercise, and Group B performed ankle exercise after hip exercise. Using a green thera-band, the dorsiflexion and plantarflexion and hip flexion and hip extension were exercised repeatedly for 15 seconds three times with a five-second rest between each set. After the exercise and measurement of one area were complete, the exercise and measurement of the other area were performed at one-day intervals. The balance ability was assessed using a Tetrax and Y-balance test and repeated three times; the best values were taken. RESULTS: In the stability index (ST) of the static balance, the hip joint exercise group (HTG) during the follow-up of normal eye open (NO) revealed notable improvement over the ankle joint exercise group (ATG), and in the follow-up of the normal eye closed (NC), the ATG showed significant improvement over the HTG. In the pillow with eye closed (PC) follow-up, the ATG showed significant improvements over the HTG. At the left (Lt) and Y-balance test (YBT), the ATG showed significant improvements in the follow-up over the HTG (p <.05). CONCLUSION: In static balance, the ATG showed significant improvement in the follow-up of NC and PC over the HTG. In the dynamic balance, the Lt. dynamic balance on the non-dominant side in the ATG showed significant improvement in the follow-up over the HTG.

• Clinical and Experimental Reproductive Medicine
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• 제50권3호
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• pp.170-176
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• 2023

Objective: Autophagy is highly active in ovariectomized mice experiencing hormone deprivation, especially in the uterine mesenchyme. Autophagy is responsible for the turnover of vasoactive factors in the uterus, which was demonstrated in anti-Müllerian hormone receptor type 2 receptor (Amhr2)-Cre-driven autophagy-related gene 7 (Atg7) knockout (Amhr-Cre/Atg7^f/f mice). In that study, we uncovered a striking difference in the amount of sequestosome 1 (SQSTM1) accumulation between virgin mice and breeder mice with the same genotype. Herein, we aimed to determine whether repeated breeding changed the composition of mesenchymal cell populations in the uterine stroma. Methods: All female mice used in this study were of the same genotype. Atg7 was deleted by Amhr2 promoter-driven Cre recombinase in the uterine stroma and myometrium, except for a triangular stromal region on the mesometrial side. Amhr-Cre/Atg7^f/f female mice were divided into two groups: virgin mice with no mating history and aged between 11 and 12 months, and breeder mice with at least 6-month breeding cycles with multiple pregnancies and aged around 12 months. The uteri were used for Western blotting and immunofluorescence staining. Results: SQSTM1 accumulation, representing Atg7 deletion and halted autophagy, was much higher in virgin mice than in breeders. Breeders showed reduced accumulation of several vasoconstrictive factors, which are potential autophagy targets, in the uterus, suggesting that the uterine stroma was repopulated with autophagy-intact cells during repeated pregnancies. Conclusion: Multiple pregnancies seem to have improved the uterine environment by replacing autophagy-deficient cells with autophagy-intact cells, providing evidence of cell mixing.

• Journal of Microbiology and Biotechnology
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• 제31권6호
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• pp.833-839
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• 2021

Bacillus subtilis CH3-5 isolated from cheonggukjang secretes a 28 kDa protease with a strong fibrinolytic activity. Its gene, aprE3-5, was cloned and expressed in a heterologous host (Jeong et al., 2007). In this study, the promoter of aprE3-5 was replaced with other stronger promoters (P_cry3A, P₁₀, P_SG1, P_srfA) of Bacillus spp. using PCR. The constructed chimeric genes were cloned into pHY300PLK vector, and then introduced into B. subtilis WB600. The P10 promoter conferred the highest fibrinolytic activity, i.e., 1.7-fold higher than that conferred by the original promoter. Overproduction of the 28 kDa protease was confirmed using SDS-PAGE and fibrin zymography. RT-qPCR analysis showed that aprE3-5 expression was 2.0-fold higher with the P10 promoter than with the original promoter. Change of the initiation codon from GTG to ATG further increased the fibrinolytic activity. The highest aprE3-5 expression was observed when two copies of the P₁₀ promoter were placed in tandem upstream of the ATG initiation codon. The construct with P10 promoter and ATG and the construct with two copies of P10 promoter in tandem and ATG exhibited 117% and 148% higher fibrinolytic activity, respectively, than that exhibited by the construct containing P10 promoter and GTG. These results confirmed that significant overproduction of a fibrinolytic enzyme can be achieved by suitable promoter modification, and this approach may have applications in the industrial production of AprE3-5 and related fibrinolytic enzymes.

• BMB Reports
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• 제53권5호
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• pp.254-259
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• 2020

Increasing evidence suggests the role of miR-449a in the regulation of tumorigenesis and autophagy. Autophagy plays an important role in the malignancy of T-cell lymphoma. However, it is still unknown whether miR-449a is associated with autophagy to regulate the malignancy of T-cell lymp homa. In this study, we for the first time demonstrated that miR-449a enhanced apoptosis of T-cell lymphoma cells by decreasing the degree of autophagy. Further, miR-449a downregulated autophagy-associated 4B (ATG4B) expression, which subsequently reduced the autophagy of T-cell lymphoma cells. Mechanistically, miR-449a decreased ATG4B protein level by binding to its mRNA 3'UTR, thus reducing the mRNA stability. In addition, studies with nude mice showed that miR-449a significantly inhibited lymphoma characteristics in vivo. In conclusion, our results demonstrated that the "miR-449a/ATG4B/autophagy" pathway played a vital role in the malignancy of T-cell lymphoma, suggesting a novel therapeutic target.

• Clinical and Experimental Pediatrics
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• 제58권7호
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• pp.267-269
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• 2015

Antithymocyte globulin (ATG) is used as an immunosuppressive treatment (IST) to deplete clonal suppressor T cells in patients with severe aplastic anemia (SAA). The depletion of suppressor T cells by ATG may affect the activation of B cells, which results in an increased risk for autoimmune conditions. A 12-year-old boy was diagnosed with idiopathic SAA. As he did not have an human leukocyte antigen-matched sibling, he was treated with rabbit ATG (3.5 mg/kg/day for 5 days) and cyclosporine. Five months later, he became transfusion independent. However, 23 months after IST, he complained of mild hand tremors, sweating, weight loss, palpitations, and goiter. Results of thyroid function tests revealed hyperthyroidism (free thyroxine, 3.42 ng/dL; thyroid stimulating hormone [TSH], <0.01 nIU/mL; triiodothyronine, 3.99 ng/mL). Results of tests for autoantibodies were positive for the antimicrosome antibody and TSH-binding inhibitory immunoglobulin, but negative for the antithyroglobulin antibody and antinuclear antibody. He was treated with methimazole, and his symptoms improved. The patient has been disease free for 39 months after IST and 9 months after methimazole treatment. This case report suggests that although rare, rabbit ATG may have implications in the pathogenesis of autoimmune hyperthyroidism. Our findings suggest that thyroid function tests should be incorporated in the routine follow-up of SAA patients treated with ATG.

• BMB Reports
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• 제52권12호
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• pp.700-705
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• 2019

The bacterial effector protein RavZ is secreted by the intracellular pathogen Legionella pneumophila and inhibits host autophagy through an irreversible deconjugation of mammalian ATG8 (mATG8) proteins from autophagosome membranes. However, the roles of the LC3 interacting region (LIR) motifs in RavZ function remain unclear. In this study, we show that a membrane-targeting (MT) domain or the LIR motifs of RavZ play major or minor roles in RavZ function. A RavZ mutant that does not bind to mATG8 delipidated all forms of mATG8-phosphatidylethanolamine (PE) as efficiently as did wild-type RavZ. However, a RavZ mutant with a deletion of the MT domain selectively delipidated mATG8-PE less efficiently than did wild-type RavZ. Taken together, our results suggest that the effects of LIR motifs and the MT domain on RavZ activity are complementary and work through independent pathways.

• 생명과학회지
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• 제19권5호
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• pp.625-632
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• 2009

결과적으로 본 연구에서 서로 다른 두 집단(RTG, ATG)의 비교시 저항성 트레이닝 집단(RTG)보다 유산소 트레이닝 집단(ATG)에서 산화질소(NO) 농도가 더 높게 나타났으며, 이는 지속적인 유산소 형태의 트레이닝이 간헐적인 저항성 형태의 트레이닝 보다 더 많은 산화질소(NO)가 생성 된다는 것을 알 수 있었다. 또한 심박수(HR)와 혈압(BP), 평균동맵압(MAP)의 감소에도 저항성 트레이닝(RT)보다는 유산소 트레이닝(AT)이 효과적이라는 것을 알 수 있었다. 그러나 유산소 트레이닝(AT)에 비해 저항성 트레이닝(RT)에서 유의하진 않았지만 산화질소(NO) 농도가 증가된 것은 주목할 만한 결과이다. 이러한 결과로 볼 때, 단기간일 지라도 혈관이완 및 자율신경계 도움을 줄 수 있는 것은 유산소 트레이닝(AT) 임을 알 수 있었고 이는 일반적으로 운동을 선택할 시(근력과 근비대를 제외한 운동) 도움을 줄 수 있다고 사료된다. 또한 추후 연구에서는 12주 이상의 장기간의 트레이닝을 통해 저항성 트레이닝(RT)이 혈관이완의 직접적인 척도인 산화질소(NO) 농도에 관한 연구가 필요시 된다.

• 분석과학
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• 제31권5호
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• pp.208-218
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• 2018

자가소화작용은 세포 내 물질들을 이중막 구조의 자가포식체를 형성하여 라이소좀(lysosome)과 융합한 후 이들을 분해시키는 과정이다. 자가소화작용은 유전적, 세포적, 생화학 수준에서 연구가 진행되어왔지만, 자가포식체의 미세구조 분석 및 정량적 분석은 체계적으로 이루어지지 않았다. 본 연구에서는 먼저, 기초 자가소화작용이 진행되는 환경과 기아상태로 자가소화작용을 유도한 환경의 세포에서 각각 자가포식체 막 형성에 중요한 역할을 하는 ATG5가 결핍된 섬유아세포와 정상 섬유아세포에서의 자가포식체의 미세구조를 비교 관찰 하였다. 정상 섬유아세포에서는 초기 패고포어, 초기 및 후기 자가포식체, 오토라이소좀과 같은 자가포식체의 단계별 미세구조를 확인한 반면, ATG5가 제거된 섬유아세포에서는 이중막 구조가 완성되지 않은 자가포식체 구조가 증가한 것을 확인하여 ATG5가 자가포식체 성숙에 중요한 역할을 함을 다시 한번 검증할 수 있었다. 본 연구를 통해 구축한 자가포식체 미세구조 분석법을 활용하여 확장된 폴리글루타민을 가지는 N-말단 헌팅틴 유전자를 발현시키는 헌팅턴병의 세포학적 모델에서 단계별 자가포식체를 분석하였다. 그 결과, 흥미롭게도 헌팅틴 단백질이 발현된 세포에서 후기 자가포식체가 증가하는 것을 확인할 수 있었으며, 이를 통해 헌팅틴 돌연변이 단백질 응집체 제거에 필요한 자가소화작용 활성화를 위해서 자가포식체와 라이소좀의 결합 단계를 촉진하는 것이 효과적일 가능성에 대해서 유추할 수 있었다. 결론적으로 전자현미경을 활용한 자가포식체의 단계별 미세구조 관찰 및 정량 분석이 다양한 인간 질병모델에 적용되고, 자가소화작용과 연관된 병리 메커니즘 연구에 기여할 수 있을 것으로 생각된다.