• BMB Reports
• /
• 제40권2호
• /
• pp.180-188
• /
• 2007

In vivo studies have demonstrated that aldosterone is an independent contributor to glomerulosclerosis. In the present study, we have investigated whether aldosterone itself mediated glomerulosclerosis, as angiotensin II (Ang II) did, by inducing cultured renal mesangial cells to produce plasminogen activator inhibitor-1 (PAI-1), and whether these effects were mediated by aldosterone-induced increase in transforming growth factor $\beta_1$ (TGF-$\beta_1$) expression and cellular reactive oxygen species (ROS) activity. Quiescent rat mesangial cells were treated by aldosterone alone or by combination of aldosterone and spironolactone, Ang II, neutralizing antibody to TGF-$\beta_1$ or antioxidant Nacetylcysteme (NAC). This study indicate that aldosterone can increase PAI-1 mRNA and protein expression by cultured mesangial cells alone, which is independent of aldosterone-induced increases in TGF-$\beta_1$ expression and cellular ROS. The effects on PAI-1, TGF-$\beta_1$ and ROS generation were markedly attenuated by spironolactone, a mineralocorticoid receptor antagonist, which demonstrate that mineralocorticoid receptor (MR) may play a role in mediating these effects of aldosterone.

• Molecules and Cells
• /
• 제40권12호
• /
• pp.906-915
• /
• 2017

Impairment of wound healing is a common problem in individuals with diabetes. Adiponectin, an adipocyte-derived cytokine, has many beneficial effects on metabolic disorders such as diabetes, obesity, hypertension, and dyslipidemia. C1q/TNF-Related Protein 9 (CTRP9), the closest paralog of adiponectin, has been reported to have beneficial effects on wound healing. In the current study, we demonstrate that CTRP9 regulates growth, differentiation, and apoptosis of HaCaT human keratinocytes. We found that CTRP9 augmented expression of transforming growth factor beta 1 ($TGF{\beta}1$) by transcription factor activator protein 1 (AP-1) binding activity and phosphorylation of p38 in a dose-dependent manner. Furthermore, siRNA-mediated suppression of $TGF{\beta}1$ reversed the increase in p38 phosphorylation induced by CTRP9. siRNA-mediated suppression of $TGF{\beta}1$ or p38 significantly abrogated the effects of CTRP9 on cell proliferation and differentiation while inducing apoptosis, implying that CTRP9 stimulates wound recovery through a $TGF{\beta}1$-dependent pathway in keratinocytes. Furthermore, intravenous injection of CTRP9 via tail vein suppressed mRNA expression of Ki67 and involucrin whereas it augmented $TGF{\beta}1$ mRNA expression and caspase 3 activity in skin of type 1 diabetes animal models. In conclusion, our results suggest that CTRP9 has suppressive effects on hyperkeratosis, providing a potentially effective therapeutic strategy for diabetic wounds.

• Laboraroty Animal Research
• /
• 제33권2호
• /
• pp.76-83
• /
• 2017

Chronic obstructive pulmonary diseases (COPD) is an important disease featured as intense inflammation, protease imbalance, and air flow limitation and mainly induced by cigarette smoke (CS). In present study, we explored the effects of $Pycnogenol^{(R)}$ (PYC, pine bark extract) on pulmonary fibrosis caused by CS+lipopolysaccharide (LPS) exposure. Mice were treated with LPS intranasally on day 12 and 26, followed by CS exposure for 1 h/day (8 cigarettes per day) for 4 weeks. One hour before CS exposure, 10 and 20 mg/kg of PYC were administered by oral gavage for 4 weeks. PYC effectively reduced the number of inflammatory cells and proinflammatory mediators caused by CS+LPS exposure in bronchoalveolar lavage fluid. PYC inhibited the collagen deposition on lung tissue caused by CS+LPS exposure, as evidenced by Masson's trichrome stain. Furthermore, transforming growth $factor-{\beta}1$ ($TGF-{\beta}1$) expression and Smad family member 2/3 (Smad 2/3) phosphorylation were effectively suppressed by PYC treatment. PYC markedly reduced the collagen deposition caused by CS+LPS exposure, which was closely involved in $TGF-{\beta}1$/Smad 2/3 signaling, which is associated with pulmonary fibrotic change. These findings suggest that treatment with PYC could be a therapeutic strategy for controlling COPD progression.

• 한국독성학회:학술대회논문집
• /
• 한국독성학회 2002년도 Molecular and Cellular Response to Toxic Substances
• /
• pp.141-141
• /
• 2002

Transforming growth factor-${\beta}$ (TGF-${\beta}$), a hormonally active polypeptide found in normal and transformed tissues, regulates cellular growth and phenotyphic plasticity. We have previously shown that H-ras, but not N-ras, induces invasive phenotype in MCF10A human breast epithelial cells.(omitted)

• Journal of Chest Surgery
• /
• 제43권4호
• /
• pp.394-398
• /
• 2010

배경: 저자들은 자연 기흉 환자의 폐 기포에서 transforming growth factor-beta 1 receptor II (TGF-${\beta}1$RII)와 transforming growth factor-beta 1 (TGF-${\beta}1$) ligand를 면역조직화학염색법으로 조사하여 상기 유전 물질이 과 발현되어 폐 기포 형성에 관여될 수 있음을 보고한 바 있다. 그러나 TGF-${\beta}1$ ligand는 혈액 내에도 존재하고 있으므로 혈액내의 TGF-${\beta}1$ ligand 양이 폐 기포 조직의 형성에도 관여가 될 수 있는 가능성 여부를 알아보기 위하여 연구를 하였다. 대상 및 방법: 자연 기흉으로 폐 기포 절제술을 실시한 환자 19명에서 폐 기포 조직과 혈액을 채취하였다. 대조군으로 25∼27세의 정상인 5명에서 혈액을 채취하였다. 획득된 폐 기포 조직은 formalin 용액에 고정하였으며 파라핀에 포매하여 $5{\sim}6{\mu}m$ 두께로 블록을 만들었으며 면역조직화학염색방법으로 염색하여 관찰하였다. 채취된 혈액에서 ELISA assay로 혈액내의 TGF-${\beta}1$의 양을 측정하였다. 결과: 19명 중 16명에서 TGF-${\beta}1$에 양성이었으며 10명에서 TGF-${\beta}1$RII에 양성의 소견을 보였다. TGF-${\beta}1$에 양성인 16명 중 9명에서 TGF-${\beta}1$RII에 양성으로 확인되었다. 강하게 염색된 부위는 폐 기포 조직과 정상 폐 조직의 경계선 부위였다. 폐 기포 조직에서 TGF-${\beta}1$과 TGF-${\beta}1$RII가 동시에 양성인 환자 9명의 혈액내의 TGF-${\beta}1$의 양은 $38.36{\pm}16.2ng/mL$이었으며 대조군은 $54.06{\pm}15ng/mL$이었다. 결론: 폐 기포를 갖는 수술 환자 군의 혈액내의 TGF-${\beta}1$의 양이 대조군보다 높지 않은 수치를 보이는 것으로 보아 혈액 내 TGF-${\beta}1$ 양은 폐 기포 형성에 직접적으로 관여될 가능성은 적고, 폐 조직에서 국소적으로 과 발현되는 TGF-${\beta}1$RII and TGF-${\beta}1$ ligand가 폐 기포 형성에 더 많이 관계하는 것으로 예상한다.

• Animal cells and systems
• /
• 제4권3호
• /
• pp.273-278
• /
• 2000

Transforming growth factor-$\beta$ (IGF-$\beta$)s are multifunctional small polypeptides synthesized in most cell types. TGF-$\beta$ exerts pivotal effects on both bone formation and resorption. In addition, increasing lines of evidence implicate TGF-$\beta$ as a potential coupling factor between these two processes during bone remodeling. In the present study, the expression form and the activation mechanism of latent-TGF-$\beta$ were investigated using specific antibodies for each isoform. TGF-$\beta$s were observed to be synthesized and accumulated in a large amount in cultured osteoblastic cells. The estimated molecular weights of intracellular TGF-$\beta$2 and -$\beta$3 were 49 and 55 kDa, respectively. Based on proteolytic digestion study and immunofluorescence observation, these precursor forms seemed to be accumulated in distinct intracellular compartments. To examine whether the internal pool of TGF-$\beta$ was possiblely regulated by external signals, their biological activites were examined in a conditioned media of this cell. Although the intact conditioned media did not contain detectable TGF-$\beta$ activity, heat-treatment or acid-activation of the conditioned media revealed significant TGF-$\beta$ activity. Furthermore, in the presence of estrogen, this activity was dramatically diminished. It is known that activation of latent TGF-$\beta$ can be achieved by different chemical and enzymatic treatments, or by incubation with certain cell types. This extracellular activation was suggested as a key step in the regulation of TGF-$\beta$ activity. In addition to these extracellular activation, this study suggests that the synthesis and intracellular processing are important regulation steps for TGF-$\beta$ action. In addition, this regulation Is specific for TGF-$\beta$ type 2, because the change was not observed in TGF-$\beta$3 in osteoblastic cell line.

• Clinical and Experimental Reproductive Medicine
• /
• 제30권1호
• /
• pp.65-75
• /
• 2003

Objectives: To investigate the role of TGF (Transforming growth factor-$\beta$) involved in the paracrinic communication during decidualization between UEC (uterine epithelial cells) and USC (uterine stromal cells), we have employed a co-culture system composed of human endometrial epithelial and stromal cells in defined hormonal conditions. Design: In the co-culture, endometrial epithelial cells cultured in the matrigel-coated cell culture insert are seeded on top of the endometrial stromal cells cultured within a collagen gel. The co-culture was maintained for 48 hours under the following hormonal conditions: progesterone dominant condition (100 nM P4 and 1 nM E2) or estrogen-dominant condition (100 nM E2 and 1 nM P4). 10 ng/ ml HGF and/or 10 ng/ml TGF-$\beta$1 are added. Methods: RT-PCR is utilized to detect mRNAs quantitatively. Enzyme-linked immunosorbent assay (ELISA) and immunohistochemical staining are utilized to detect proteins in the tissue. Results: Prolactin mRNA is expressed in the co-cultured stromal cells under the progesterone dominant condition. TGF-$\beta$1 and its receptors are expressed in both the co-cultured epithelial and stromal cells irrespective of the steroid present, which is in contrast with no or negligible expression of TGF-$\beta$1 or its receptor in cells separately cultured. Both estrogen and progesterone significantly elevate the concentration of hepatocyte growth factor (HGF) in the conditioned medium of the co-culture with the value of 4, 325 pg/ml in E2-dominant and 2, 000 pg/ml in P4-dominant condition compare to 150 pg/ml in no hormone. In separately cultured stromal cells, administration of HGF induces the expression of TGF receptor 1 in both hormonal conditions, but induction of TGF receptor 2 is only manifest in the P4-dominant condition. Administration of TGF-$\beta$ and HGF directly induce the decidualization marker prolactin mRNA in separately cultured stromal cells. Conclusion: It is likely that steroid hormones induces prolactin mRNA indirectly by promoting the cell to cell communication between the stromal and the epithelial cells. TGF-$\beta$ and HGF are two possible paracrine mediators in the human endometrial decidualization.

• 생명과학회지
• /
• 제18권4호
• /
• pp.461-466
• /
• 2008

Transforming growth $factor-{\beta}$ ($TGF-{\beta}$)에 의해 유도되는 세포사멸 과정은 정상 조직에서 손상 받은 조직이나 비정상 적인 조직을 제거하는데 중요한 역할을 담당한다. Gadd45b는 p38 kinase를 활성화시킴으로 $TGF-{\beta}$에 의해 유도되는 세포사멸 과정을 매개한다고 알려져 있다. 본 연구에서는 $TGF-{\beta}$에 의해 세포사멸이 일어나는 EpH4 세포에서 Gadd45b 유전자의 발현이 $TGF-{\beta}$에 의해 촉진됨을 보여주었다. 어떠한 기작으로 $TGF-{\beta}$에 의해 Gadd45b 유전자의 발현이 촉진되는지 알아보기 위해 Gadd45g 유전자의 5'-flanking region을 cloning하였으며, EpH4 세포에서 $TGF-{\beta}$에 의해 그 promoter activity가 증가함을 확인하였다. 여러 가지 deletion mutants를 제조하여 promoter activity를 조사한 결과 전사 개시점으로부터 220 bp upstream 부위 에 promoter activity에 필수적인 sequence가 존재함을 확인하였다. 또한 $TGF-{\beta}$에 의한 Gadd45b 유전자의 promoter activity에 Smad2, Smad3, 그리고 Smad4가 중요한 기능을 담당함도 확인하였다. 마지막으로 ras 유전자가 도입되어 $TGF-{\beta}$에 의한 세포사멸이 억제되어있는 EpRas 세포에서 $TGF-{\beta}$에 의한 Gadd45b 유전자의 발현을 확인한 결과 EpRas 세포에서 $TGF-{\beta}$에 의한 Gadd45b 유전자의 발현이 억제됨을 확인하였다. 이러한 결과는 Gadd45b 유전자가 EpH4 세포에서 $TGF-{\beta}$에 의한 세포사멸을 유도하는데 중요한 기능을 담당할 가능성이 높음을 의미하는 것이다.

• Clinical and Experimental Reproductive Medicine
• /
• 제47권2호
• /
• pp.108-113
• /
• 2020

Objective: Endometrial fibrosis, the primary pathological feature of intrauterine adhesion, may lead to disruption of endometrial tissue structure, menstrual abnormalities, infertility, and recurrent pregnancy loss. At present, no ideal therapeutic strategy exists for this fibrotic disease. Eupatilin, a major pharmacologically active flavone from Artemisia, has been previously reported to act as a potent inducer of dedifferentiation of fibrotic tissue in the liver and lung. However, the effects of eupatilin on endometrial fibrosis have not yet been investigated. In this study, we present the first report on the impact of eupatilin treatment on transforming growth factor beta (TGF-β)-induced endometrial fibrosis. Methods: The efficacy of eupatilin on TGF-β-induced endometrial fibrosis was assessed by examining changes in morphology and the expression levels of fibrosis markers using immunofluorescence staining and quantitative real-time reverse-transcription polymerase chain reaction. Results: Eupatilin treatment significantly reduced the fibrotic activity of TGF-β-induced endometrial fibrosis in Ishikawa cells, which displayed more circular shapes and formed more colonies. Additionally, the effects of eupatilin on fibrotic markers including alpha-smooth muscle actin, hypoxia-inducible factor 1 alpha, collagen type I alpha 1 chain, and matrix metalloproteinase-2, were evaluated in TGF-β-induced endometrial fibrosis. The expression of these markers was highly upregulated by TGF-β pretreatment and recovered to the levels of control cells in response to eupatilin treatment. Conclusion: Our findings suggest that suppression of TGF-β-induced signaling by eupatilin might be an effective therapeutic strategy for the treatment of endometrial fibrosis.

• KSBB Journal
• /
• 제14권1호
• /
• pp.9-16
• /
• 1999

Transforming growth factor $\beta$1(TGF-$\beta$1)은 여러 가지 생물학적 활성을 가지는 관계로 의학적 치료제로서 사용될 가능성이 크다. 본 연구에서는 혈소판추출, 젤여과, 양이온교환 크로마토그래피 및 역상 HPLC등 네 단계의 정제과정으로 이루어져 있는 정제공정을 이용한 TGF-$\beta$1을 값싸고 효울적으로 정제하였다. 이 과정을 거쳐 최종적으로 얻어진 TGF-$\beta$1은 비환원조건하에서 SDS-PAGE를 행한 결과 구매된 TGF-$\beta$1 표준품과 일치한 위치에서 한 개의 band가 관찰되어 순수하다는 것을 확인하였으며 또한 이것이 Westem blot를 통하여 TGF-$\beta$1 항체와 결합하는 것으로부터 TGF-$\beta$1임을 확인하였다 또한, mink lung epithelial cell line 을 이용한 성장저해 실험을 통해 정제된 TGF-$\beta$1이 구매되TGF-$\beta$1 표준품보다 조금 높은 활성을 가지는 것을 확인하였다 최종적으로 농축혈소판 10단위로부터 약 3.7$\mu$g의 정제된 TGF-$\beta$1이 얻어져 그 최종수율은 약 21%였다.