• The Korean Journal of Physiology and Pharmacology
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• 제1권4호
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• pp.435-443
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• 1997

We employed the whole-cell patch clamp technique to investigate the effects of arachidonic acid (AA) on barium inward current through the L-type calcium channels ($I_{Ba}$) and on osmotic stretch-induced increase of $I_{Ba}$ in guinea-pig antral gastric myocytes. Under isosmotic condition, AA inhibited $I_{Ba}$ in a dose-dependent manner to $91.1{\pm}1.4,\;72.0{\pm}3.2,\;46.0{\pm}1.8,\;and\;20.3{\pm}2.3%$ at 1, 5, 10, 30 mM, respectively. The inhibitory effect of AA was not affected by 10 ${\mu}M$ indomethacin, a cyclooxygenase inhibitor. Other unsaturated fatty acids, linoleic acid (LA) and oleic acid (OA) were also found to suppress $I_{Ba}$ but stearic acid (SA), a saturated fatty acid, had no inhibitory effect on $I_{Ba}$. The potency sequence of these inhibitory effects was AA ($79.7{\pm}2.3%$) ＞ LA ($43.1{\pm}2.7%$) ＞ OA ($14.2{\pm}1.1%$) at 30 ${\mu}M$. On superfusing the myocyte with hyposmotic solution (214 mOsm) the amplitude of $I_{Ba}$ at 0 mV increased ($38.0{\pm}5.5%$); this increase was completely blocked by pretreatment with 30 mM AA, but not significantly inhibited by lower concentrations of AA (1, 5 and 10 ${\mu}M$) (P＞0.05). Unsaturated fatty acids shifted the steady-state inactivation curves of $I_{Ba}$ to the left; the extent of shift caused by AA was greater than that caused by LA. The activation curve was not affected by AA or LA. The results suggest that AA and other unsaturated fatty acids directly modulate L-type calcium channels and AA might modulate the hyposmotic stretch- induced increase of L-type calcium channel current in guinea-pig gastric smooth muscle.

• Journal of Chest Surgery
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• 제41권2호
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• pp.160-169
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• 2008

배경: 혈관질환의 수술에 사용되는 인공 도관의 막힘과 문합부위의 좁아짐 등을 개선하기 위한 방법으로 조직공학적인 방법과 자가 세포를 이용한 인공혈관의 제작이 대안으로 대두되고 있다. 저자들은, 생흡수성이 있는 고분자 폴리머 지지체와 자가 골수세포를 이용한 인공혈관으로 생체실험을 시행하였다. 대상 및 방법: 생분해성 고분자 재료인 poly (lactide-co-${\varepsilon}$-caprolactone) (PLCL)과 poly(glycolic acid) (PGA) fiber로 혈관용 지지체를 제작한 후, 피실험 동물의 골수를 채취하여 혈관 내피 세포와 평활근 세포로 분열시켜 배양한 후 혈관 지지체위에 이식하였다. 만들어진 인공 혈관을 잡견의 복부대동맥에 이식한 후 3주 후에 혈관 조영술을 시행하고, 안락사 후에 혈관을 제거하여 조직학적 검사를 시행하였다. 결과: 6마리의 잡견 중 2마리에서 수술 후 10일에 혈관 지지체의 균열에 의한 대량 출혈로 사망하였다. 나머지 4마리의 잡견은 수술 후 3주까지 생존하였으며, 혈관 조영술상 혈관의 막힘이나 좁아짐은 발견되지 않았다. 인공 혈관의 내면은 작은 혈전들이 붙어 있었으며, 조직학 검사에서 정상 혈관과 유사한 3층의 구조를 나타내었다. 또한 면역화학 검사에서 혈관 내피세포와 혈관 평활근 세포가 재생된 것을 확인하였다. 결론: 고분자 폴리머와 자가 골수세포를 이용한 인공혈관은 생체 내에서 정상혈관과 유사한 모양으로 재생이 가능함을 보여주었다. 그러나, 동맥압력에 견디기 위해 혈관 지지체의 물성에 대한 개량과 충분한 양의 혈관 세포를 얻기 위한 연구가 더 필요할 것으로 생각된다.

• Molecules and Cells
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• 제27권2호
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• pp.175-181
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• 2009

The myosin light chain kinase (MYLK) gene encodes both smooth muscle and nonmuscle cell isoforms. Recently, polymorphisms in MYLK have been reported to be associated with several diseases. To examine the genetic effects of polymorphisms on the risk of asthma and related phenotypes, we scrutinized MYLK by re-sequencing/genotyping and statistical analysis in Korean population (n = 1,015). Seventeen common polymorphisms located in or near exons, having pairwise $r^2$ values less than 0.25, were genotyped. Our statistical analysis did not replicate the associations with the risk of asthma and log-transformed total IgE levels observed among African descendant populations. However, two SNPs in intron 16 (+89872C> G and +92263T> C), which were in tight LD (|D'| = 0.99), revealed significant association with log-transformed blood eosinophil level even after correction multiple testing ($P=0.002/P^{corr}=0.01$ and $P=0.002/P^{corr}=0.01$, respectively). The log-transformed blood eosinophil levels were higher in individuals bearing the minor alleles for +89872C> G and +92263T> C than in those bearing other allele. In additional subgroup analysis, the genetic effects of both SNPs were much more apparent among asthmatic patients and atopic asthma patients. Among atopic asthma patients, the log-transformed blood eosinophil levels were proportionally increased by gene-dose dependent manner of in both +89872C> G and +92263T> C(P = 0.0002 and P = 0.00007, respectively). These findings suggest that MYLK polymorphisms might be among the genetic factors underlying differential increases of blood eosinophil levels among asthmatic patients. Further biological and/or functional studies are needed to confirm our results.

• The Korean Journal of Physiology
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• 제24권2호
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• pp.293-303
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• 1990

The contractile action of barium $(Ba^{2+})$ was investigated in the arterial strip of rabbit renal artery. The helical strip of isolated renal artery was immersed in the Tris-buffered Tyrode's solution equilibrated with 100% $O_2$ at $37^{\circ}C$ and its isometric tension was measured. $Ba^{2+}-induced$ contraction of arterial strip was dose-dependent and its maximal tension corresponded to $92.1{\pm}4.5%$ of tension by $K^+(100\;mM)$. $Ba^{2+}-induced$ contraction did not show the tachyphylactic phenomenon in the normal Tyrode's solution. $Ba^{2+}$ induced the tonic contraction in the $Ca^{2+}-free$ tyrode's solution and that was increased by the extracellula addition of $Ca^{2+}$. During the repeated exposure of the same dose of $Ba^{2+}\;(10\;mM)$ in the $Ca^{2+}-free$ Tyrode's solution, $Ba^{2+}-induced$ contraction was progressively decreased. Even though the intracellular NE-and caffeine-sensitive $Ca^{2+}$ was depleted, $Ba^{2+}$ induced the tonic contraction. After the pretreatment of lanthnum or verapamil, $Ba^{2+}$ did not induce contraction. $Ba^{2+}-induced$contraction was suppressed by extracellular $K^+$ in the normal Tyrode's solution and that was dependent on $K^+$ concentration. Suppressive effect of $K^+\;(14\;mM)$ on the $Ba^{2+}-induced$ contraction was also dependent on the intracellular $Ca^{2+}$ concentration. From the above resuts, it is suggested that $Ba^{2+}$ activate indirectly the contractile process by promoting the mobilization of intracellular $Ca^{2+}$ and the influx of extracellular $Ca^{2+}$. It is also suggested that action of $Ba^{2+}$ on the $Ca^{2+}-activated$ $K^+$ channel can result in the depolarization of cell membrane in the rabbit renal artery.

• 한국산학기술학회논문지
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• 제20권8호
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• pp.477-484
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• 2019

포유류에 존재하는 Collectin-Placenta 1 (CL-P1)을 포함한 여러 종류의 scavenger 수용체는 주로 내피 세포, 대식 세포 및 평활근 세포 표면에 발현되는 분자이다. 이들 분자는 산화 된 저밀도 지질 단백질 (oxLDL)에 결합하여 처리 할 수 있는 세포 표면 당 단백질이다. 이들 분자 중 케르세틴이 CL-P1 활성화에 어떤 영향을 미치는가를 확인하였다. 케르세틴은 산화 반응을 담당하는 자유 라디칼의 제거제 역할을 하여 산화를 중지시키는 항산화제로 알려져 있다. 본 논문에서는 마우스 CL-P1 유전자 promoter 부분의 전사 시작 점부터 -500 번째 염기까지의 단편을 DNA 중합효소를 이용하여 클로닝 하였다. 그 후에 대식세포 계열인 RAW264.7 및 섬유아세포계열의 NIH3T3 세포에 도입하여 케르세틴이 CL-P1 유전자 발현에 어떠한 영향을 미치는지에 대한 연구를 하였다. 이 부위에는 세포주기 조절 인자인 E2F 결합부위를 비롯해서 여러 종류의 전사 인자가 결합하는 염기서열이 다수 위치하고 있다. 이러한 500염기 단편을 pGL4.10 기본 벡터 및 프로모터에 연결시킨 후 세포에 도입시켰다. 그리고 배양 중에 케르세틴을 처리하여 유전자 발현양을 형광 색소 발현기법으로 측정하였다. 그 결과 유전자 발현이 시작되는 앞쪽 부분의 -250에서 -350사이의 염기들이 CL-P1 단백질을 만드는데 중요하다는 것을 확인하였다. 그 중에서도 E2F결합 부위가 결정적인 것 이라는 것을 DNA 돌연변이 실험을 통해 확인 하였다. 또한 부착 세포인 RAW264.7 배양액에 케르세틴을 첨가 한 결과, 배양용기 표면에서 탈락하는 현상을 확인하였다. 즉 발현된 CL-P1단백질이 케르세틴에 의해 세표 표면의 부착 분자에도 영향을 주는 것을 확인하였다.

• Journal of Veterinary Science
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• 제21권2호
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• pp.26.1-26.14
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• 2020

Pancreatic ductal adenocarcinoma is a lethal cancer type that is associated with multiple gene mutations in somatic cells. Genetically engineered mouse is hardly applicable for developing a pancreatic cancer model, and the xenograft model poses a limitation in the reflection of early stage pancreatic cancer. Thus, in vivo somatic cell gene engineering with clustered regularly interspaced short palindromic repeats is drawing increasing attention for generating an animal model of pancreatic cancer. In this study, we selected Kras, Trp53, Ink4a, Smad4, and Brca2 as target genes, and applied Campylobacter jejuni Cas9 (CjCas9) and Streptococcus pyogens Cas9 (SpCas9) for developing pancreatic cancer using adeno associated virus (AAV) transduction. After confirming multifocal and diffuse transduction of AAV2, we generated SpCas9 overexpression mice, which exhibited high double-strand DNA breakage (DSB) in target genes and pancreatic intraepithelial neoplasia (PanIN) lesions with two AAV transductions; however, wild-type (WT) mice with three AAV transductions did not develop PanIN. Furthermore, small-sized Cjcas9 was applied to WT mice with two AAV system, which, in addition, developed high extensive DSB and PanIN lesions. Histological changes and expression of cancer markers such as Ki67, cytokeratin, Mucin5a, alpha smooth muscle actin in duct and islet cells were observed. In addition, the study revealed several findings such as 1) multiple DSB potential of AAV-CjCas9, 2) peri-ductal lymphocyte infiltration, 3) multi-focal cancer marker expression, and 4) requirement of > 12 months for initiation of PanIN in AAV mediated targeting. In this study, we present a useful tool for in vivo cancer modeling that would be applicable for other disease models as well.

• 한방재활의학과학회지
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• 제33권3호
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• pp.47-65
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• 2023

Objectives We evaluated the wound healing effects of Dangguijakyak-san (DJ) using C57BL/6 mice that were generated open wound. Methods The study was conducted with seven C57BL/6 mice assigned to each group, divided into the normal group, control group, vitamin E group, DJ low-dose group, DJ high-dose group. We measured total polyphenol, flavonoid contents, the size of the wound, liver function, pro-inflammatory cytokine activity in serum, inflammation-related proteins, adhesion molecules and chemokine proteins, collagen-related proteins in skin tissue and histopathological changes by H&E and Masson's staining. Results DJ treatment significantly reduced the area of the wound compared to the control group. Also, inflammatory cytokines were reduced and the expression of anti-inflammatory-related factors (interleukin-4 [IL-4] and IL-10) was significantly increased in the DJ treatment group. We identified that DJ treatment inhibits both pathways of inflammation, the mitogen-activated protein kinases and nuclear factor-κB pathway. Moreover, the protein expressions of Sirt1 (sirtuin 1), MCP-1 (monocyte chemoattractant protein 1), ICAM-1 (intercellular adhesion molecule 1), and VCAM-1 (vascular cell adhesion molecule 1) were decreased by DJ administration. Also, the expression of α-smooth muscle actin and collagen type I alpha 1, collagen-related proteins, that help skin recovery was significantly increased in the DJ treatment group. Histopathologically, a relatively thin epithelial layer could be observed in the DJ administration group, as well as an increase in fibroblasts and collagen fibers. Conclusions These data suggest that DJ treatment is effective in wound healing, suppressing inflammatory proteins, increasing skin repair factors and improving histopathological changes caused by wounds.

• 한방안이비인후피부과학회지
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• 제36권4호
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• pp.30-50
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• 2023

Objectives : To investigate the active compounds and therapeutic mechanisms of Atractylodes Lancea(Thunb.) D.C. and Magnolia Officinalis Rehder et Wilson in the treatment of dermatitis accompanied by pruritus, as well as their potential to complement or replace standard drugs. Methods : We conducted the network pharmacological analysis. We selected effective ingredients among the active compounds of research target herbs. Then we explore pathway/terms of the common target proteins among research target herbs, fexofenadine and disease. Results : We selected 9 active compounds are selected from Atractylodes lancea and identified 231 target proteins. Among them, 74 proteins are associated with inflammatory skin diseases that cause pruritus. These proteins are involved in various pathways including, 'Nitric-oxide synthase regulator activity', 'Hydroperoxy icosatetraenoate dehydratase activity, Aromatase activity', 'RNA-directed DNA polymerase activity', 'Arachidonic acid metabolism', 'Peptide hormone processing', 'Chemokine binding' and 'Sterol biosynthetic process'. Additionally, coregenes are involved in 'IL-17 signaling pathway'. Similarly, we selected 2 active compounds from Magnolia officinalis and identified 133 target proteins. Among them, 33 proteins are related to inflammatory skin diseases that cause pruritus. These proteins are primarily involved in 'Vascular associated smooth muscle cell proliferation' and 'Arachidonic acid metabolism'. There is no significant difference between the pathways in which coregenes are involved. Conclusions : It is expected that Atractylodes Lancea will be able to show direct or indirect anti-pruritus and anti-inflammatory effects on skin inflammation accompanied pruritus through suppressing inflammation and protecting skin barrier. Meanwhile, it is expected that Magnolia Officinalis will only be able to show indirect anti-inflammation effects. Therefore, Atractylodes Lancea and fexofenadine are believed to complement each other, whereas Magnolia Officialinalis is expected to provide supplementary support on skin disease.

• International Journal of Stem Cells
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• 제16권1호
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• pp.52-65
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• 2023

Background and Objectives: Epithelial-Mesenchymal transition (EMT) is one of the origins of myofibroblasts in renal interstitial fibrosis. Mesenchymal stem cells (MSCs) alleviating EMT has been proved, but the concrete mechanism is unclear. To explore the mechanism, serum-free MSCs conditioned medium (SF-MSCs-CM) was used to treat rat renal tubular epithelial cells (NRK-52E) fibrosis induced by transforming growth factor-β1 (TGF-β1) which ameliorated EMT. Methods and Results: Galectin-3 knockdown (Gal-3 KD) and overexpression (Gal-3 OE) lentiviral vectors were established and transfected into NRK-52E. NRK-52E fibrosis model was induced by TGF-β1 and treated with the SF-MSCs-CM for 24 h after modelling. Fibrosis and autophagy related indexes were detected by western blot and immunocytochemistry. In model group, the expressions of α-smooth muscle actin (α-SMA), fibronectin (FN), Galectin-3, Snail, Kim-1, and the ratios of P-Akt/Akt, P-GSK3β/GSK3β, P-PI3K/PI3K, P-mTOR/mTOR, TIMP1/MMP9, and LC3B-II/I were obviously increased, and E-Cadherin (E-cad) and P62 decreased significantly compared with control group. SF-MSCs-CM showed an opposite trend after treatment compared with model group. Whether in Gal-3 KD or Gal-3 OE NRK-52E cells, SF-MSCs-CM also showed similar trends. However, the effects of anti-fibrosis and enhanced autophagy in Gal-3 KD cells were more obvious than those in Gal-3 OE cells. Conclusions: SF-MSCs-CM probably alleviated the EMT via inhibiting Galectin-3/Akt/GSK3β/Snail pathway. Meanwhile, Gal-3 KD possibly enhanced autophagy via inhibiting Galectin-3/Akt/mTOR pathway, which synergistically ameliorated renal fibrosis. Targeting galectin-3 may be a potential target for the treatment of renal fibrosis.

• Journal of Chest Surgery
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• 제36권12호
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• pp.887-893
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• 2003

외부 자극에 의해 세포 내 $Ca^{2+}$이 증가하면 $K^{+}$이 유출되는 기전을 통해 세포 외 $K^{+}$이 증가하는데, 이 $K^{+}$ 의 증가가 혈관 수축에 미치는 영향을 규명하고자 쥐 대동맥 혈관내피세포를 이용해 실험을 시행하였다. 대상 및 방법: 세포 외 $K^{+}$ 농도를 증가시키거나, 혈관 내피세포의 제거, nitric oxide 생성 억제제인 L-NAME (N-nitro-L-arginine methyl ester)의 투여, $Na^{+}$- $K^{+}$ pump 억제제인 Ouabain, $Na^{+}$-C $a^{2+}$ exchanger 억제제인 N $i^{2+}$의 투여 등 조건을 달리하며, 막전압고정법을 이용, $Ca^{2+}$ 변화와 여러 이온 전류 변화를 측정해 혈관의 수축성을 알아보았다. 결과: 세포외 $K^{+}$ 농도를 6에서 12 mM 증가시켜도 norepinephrine에 의한 혈관의 수축성에는 변화가 없었고, 12 mM 상으로 증가시키면 평활근이 수축하기 시작하였다. Acetylcholine (ACh)에 의해 유발된 내피세포 의존성 이완은 세포 외 $K^{+}$ 농도를 6에서 12 mM로 증가시키면 억제되었으며, 혈관내피세포를 제거하거나 L-NAME을 투여하는 경우에 ACh에 의한 이완은 일어나지 않았다. 배양한 쥐 대동맥 내피세포에서는 ATP혹은 ACh에 의해 세포 내 $Ca^{2+}$이 증가하였으며, 세포 내 $Ca^{2+}$ 증가가 정점에 이른 후 세포 외 $K^{+}$을 6에서 12 mM로 증가시키면 세포 내 $Ca^{2+}$이 농도 의존적으로 감소하였으나 다시 6 mM로 감소시키면 세포 내 $Ca^{2+}$이 증가하였다. 또한 세포 외 $K^{+}$ 증가에 의한 내피세포 의존성 이완효과는 Ouabain과 N $i^{2+}$에 의하여 억제되었다. 걸론 세포 외 $K^{+}$의 증가는 저항혈관 평활근은 이완시키며, 혈관내피세포 $Ca^{2+}$을 감소시켜 내피세포 의존성 이완을 억제하는데 이는 $Na^{+}$- $K^{+}$ pump와 $Na^{+}$-C $a^{2+}$exchanger를 활성화시켜 일어나는 것으로 생각된다.