• Journal of Embryo Transfer
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• v.29 no.1
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• pp.73-81
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• 2014

The in vitro production of porcine embryos was essential to increase of blastocyst development rate and select of high quality blastocyst in early stage. There were a lot of reports about in vitro porcine embryo development, but there was no report about the selection of high quality embryos. Therefore, in this study, we investigated the effect of vitamin $K_1$ (vit $K_1$) on the development and survival rate of porcine in vitro fertilized embryos. When vit $K_1$ was treated for 24 hr at day 1 in vitro culture, blastocyst development rate in the control group ($35.5{\pm}3.2%$) was significantly lower compared to $1.0{\mu}M$, $3.0{\mu}M$, or $6.0{\mu}M$ groups ($14.5{\pm}4.3$, 0.0, or 0.0%; p<0.05). The survival rates of blastocysts at day 8 in $1.0{\mu}M$, $3.0{\mu}M$ or $6.0{\mu}M$ of vit $K_1$ treated groups ($22.2{\pm}2.9$, 0.0 or 0.0%) were significantly lower than that of the control group ($31.8{\pm}2.6%$; p<0.05). We were added at $1.0{\mu}M$, $3.0{\mu}M$ or $6.0{\mu}M$ vit $K_1$ for different durations of time at day 1 in vitro culture. The development rate and survival rate in the group of $1.0{\mu}M$ vit $K_1$ for 6 hr was $26.5{\pm}2.9%$ and $47.2{\pm}2.8%$, respectively, which were differed significantly in the group of 12 hr (p<0.05). In the group of $3.0{\mu}M$ vit $K_1$, the blastocyst development in control group was $36.4{\pm}3.1%$ but, the survival rate $41.7{\pm}3.2%$ in the group of 3.0 hr was significantly higher than that of the control group (p<0.05). In the group of $6.0{\mu}M$ vit $K_1$, the control group's the blastocyst development was $32.0{\pm}2.8%$ and the 0.5 hr supplement group's survival rates was $42.9{\pm}1.8%$ higher than other groups. We added vit $K_1$ at day 1, day 2, day 4 and day 6 of in vitro culture, on the based the results of supplemented concentration and duration. In the group of $1.0{\mu}M$ 6.0 hr addition, the blastocyst development rate of day 4 and the survival rate of day 2 were the highest in each group. In the groups of $3.0{\mu}M$ 3.0 hr addition or $6.0{\mu}M$ 0.5 hr addition, the blastocyst development ($59.5{\pm}4.1%$ and $50.0{\pm}3.6%$) and survival rates ($72.7{\pm}5.4%$ and $79.2{\pm}4.0%$) on day 4 were significantly higher than that of control and other experiment groups (p<0.05). Meanwhile, the number of cells in blastocysts that produced by vit $K_1$ supplementation was $53.4{\pm}5.8$, $49.4{\pm}3.8$ and $51.5{\pm}4.5$ respectively, which were significantly higher than that of $40.2{\pm}2.3$ in the control group (p<0.05). There was no difference of the number of apoptotic cells between control and experiment groups. In addition, gene expression of survival blastocyst, the Bax mRNA expression was similar between the control and the experiment groups. However, Bcl-xL mRNA expression's in the group of $6.0{\mu}M$ 0.5 hr on day 4 was highest among control and experiment groups (p<0.05). In this study suggested that the control of concentration, duration and time was effective on the survival and cell number of porcine blastocyst derived from in vitro. We are not know what the exact reasons of the effect of vit $K_1$ on embryo development and need to fur ther study. However, vit $K_1$ might be using the selection of high quality porcine blastocyst.

• Tuberculosis and Respiratory Diseases
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• v.45 no.4
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• pp.861-869
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• 1998

Backgrounds: The injury of a tissue results in the infalmmation, and the imflammed tissue is replaced by the normal parenchymal cells during the process of repair. But, constitutional or repetitive damage of a tissue causes the deposition of collagen resulting in the loss of its function. These lesions are found in the lung of patients with idiopathic pulmonary fibrosis, complicated fibrosis after diffuse alveolar damage (DAD) and inorganic dust-induced lung fibrosis. The tissue from lungs of patients undergoing episodes of active and/or end-stage pulmonary fibrosis shows the accumulation of inflammatory cells, such as mononuclear cells, neutrophils, mast cells and eosinophils, and fibroblast hyperplasia. In this regard, it appears that the inflammation triggers fibroblast activation and proliferation with enhanced matrix synthesis, stimulated by inflammatory mediators such as interleukin-1 (IL-1) and/or tumor necrosis factor (TNF). It has been well known that TGF-$\beta$ enhance the proliferation of fibroblasts and the production of collagen and fibronectin, and inhibit the degradation of collagen. In this regard, It is likely that TGF-$\beta$ undergoes important roles in the pathogenesis of pulmonary fibrosis. Nevertheless, this single cytokine is not the sole regulator of the pulmonary fibrotic response. It is likely that the balance of many cytokines including TGF-$\beta$, IL-1, IL-6 and TNF-$\alpha$ regulates the pathogenesis of pulmonary fibrosis. In this study, we investigate the interaction of TGF-$\beta$, IL-1$\beta$, IL-6 and TNF-$\alpha$ and their effect on the proliferation of fibroblasts. Methods: We used a human fibroblast cell line, MRC-5 (ATCC). The culture of MRC-5 was confirmed by immunofluorecent staining. First, we determined the concentration of serum in cuture medium, in which the proliferation of MRC-5 is supressed but the survival of MRC-5 is retained. Second, we measured optical density after staining the cytokine-stimulated cells with 0.5% naphthol blue black in order to detect the effect of cytokines on the proliferation of MRC-5. Result: In the medium containing 0.5% fetal calf serum, the proliferation of MRC-5 increased by 50%, and it was maintained for 6 days. IL-1$\beta$, TNF-$\alpha$ and IL-6 induced the proliferation of MRC-5 by 45%, 160% and 120%, respectively. IL-1$\beta$ and TNF-$\alpha$ enhanced TGF-$\beta$-induced proliferation of MRC-5 by 64% and 159%, but IL-6 did not affect the TGF-$\beta$-induced proliferation. And lNF-$\alpha$-induced proliferation of MRC-5 was reduced by IL-1$\beta$ in 50%. TGF-$\beta$, TNF-$\alpha$ and both induced the proliferation of MRC-5 to 89%, 135% and 222%, respectively. Conclusions: TNF-$\alpha$, TGF-$\beta$ and IL-1$\beta$, in the order of the effectiveness, showed the induction of MRC-5 proliferation of MRC-5. TNF-$\alpha$ and IL-1$\beta$ enhance the TGF-$\beta$-induced proliferation of MRC-5, but IL-6 did not have any effect TNF-$\alpha$-induced proliferation of MRC-5 is diminished by IL-1, and TNF-$\alpha$ and TGF-$\beta$ showed a additive effect.