SDF-1과 HGF 에 의한 인체 골수와 재대혈에서 분리배양된 중간엽 줄기세포의 이동

손보라 충북대학교 의과대학 충북대학교 의학연구소 2005 忠北醫大學術誌 Vol.15 No.2

연구 목적: 손상된 장기와 조직을 재생 시키기 위한 세포 치료법들 중 중간염 줄기세포(msenchymal stem cell: MSC)를 이용한 방법이 주목받고 있다. 그러나 이러한 세포가 손상이 일어난 곳으로 이동하고 생착하는 일련의 과정에 대해서는 알려진 바가 적다. 최근에 stromal-derived factor (SDF-1)와 hepatocyte growth factor (HGF)가 조직이나 장기가 손상되는 동안 증가하는 것으로 보고되고 있어, 본 실험에서 중간엽 줄기세포들이 SDF-1과 HGF에 반응하여 이동할 수 있는 지를 알아 보고자 하였다. 대상 및 방법: 인제 골수와 재대혈에서 분리 배양된 중간염 줄기세포를 체외에서 계대배양하며 CXCR4 (SDF-1의 수용체)dhk c-met (HGF의 수용체)dml 발현여부를 역전사 중합효소반응으로 확인하고, 기능적으로 활성화 되어 있는 지를 알아보기 위해 초기(3차), 중기(10차)와 후기(15차)의 계대배양 세포주들을 대상으로 SDF-1과 HGF에 대한 trans-matrigel chemoinvasion assay를 시행하였다. 결과: 인체 골수와 재대혈에서 분리 배양된 중간엽 줄기세포 모두 일차부터 15차까지의 계대배양 세포주들에서 CXCR4와 c-met의 발현을 확인하였다. Trans-matrigel chemoinvasion assay를 시행한 결과, 후기 계대배양 세포주들까지 SDF-1과 HGF에 대해 반응하여 이동함을 관찰하였다. 결론: SDF-1과 HGF가 인체 골수와 재대혈에서 분리배양된 중간엽 줄기세포의 이동과정에 중요한 역할을 담당하는 것으로 보인다. Purpose: Human mesenchymal stem cells (MSC) are increasingly being considered in cell-based therapeutic strategies for regeneration of various organs/tissues. However, the signals required for their homing and recruitment to injured sites are not yet fully understood. Because stromal-derived factor (SDF)-l and hepatocyte growth factor (HGF) become upregulated during tissue/organ damage, this study examined whether these factors chemoattract ex vivo-expanded MSC derived from bone marrow (BM) and umbilical cord blood (CB). Materials and Methods: This study investigated the expression by MSC of CXCR4 and c-met, the cognate receptors of SDF-1 and HGF, respectively, and their functionality after early and late passages of MSC. We maintained expanded BM- or CB-derived MSC for up to 15-18 passages with monitoring of the expression of functional CXCR4 and cmet. Results: For up to 15-18 passages, both BM- and CB-derived MSC i) express CXCR4 and c-met receptors and are strongly attracted by SDF-1 or HGF gradients, ii) are chemomvasive across the reconstituted basement membrane Matrigel. Conclusion: These results suggest that the SDF-1-CXCR4 and HGF-c-met axes may be involved in recruitment of expanded MSC to damaged tissues.

치주염 원인균 LPS-PG로 유도된 인체 치은섬유아세포에서 연뿌리 추출물에 대한 항염증 및 항산화 효과

이영경,김철환,정대원,이기원,오영택,김정일,정진우 한국자원식물학회 2022 한국자원식물학회지 Vol.35 No.5

Abstract - Gingival inflammation is one of the main causes that can be related to various periodontal diseases. Human gingival fibroblast (HGF) is the major constituent in periodontal connective tissue and secretes various inflammatory mediators, such as nitric oxide (NO) and prostaglandin E2 (PGE2), upon lipopolysaccharide stimulation. This study is aimed at investigating the anti-inflammatory and antioxidative activities of Lotus Root extract (LRE) in Porphyromonas gingivalis derived lipopolysaccharide (LPS-PG)-stimulated HGF-1 cells. The concentration of NO and PGE2, as well as their responsible enzymes, inducible NO synthase (iNOS), and cyclooxygenase-2 (COX-2), was analyzed by Griess reaction, ELISA, and western blot analysis. LPS-PG sharply elevated the production and protein expression of inflammatory mediators, which were significantly attenuated by LRE treatment in a dose-dependent manner. LRE treatment also suppressed activation of Toll-like receptor 4 (TLR4)/myeloid differentiation primary response gene 88 (MyD88) and nuclear factor-κB (NF-κB) in LPS-PG-stimulated HGF-1 cells. In addition, one of phase II enzyme, NAD(P)H quinone dehydrogenase (NQO)-1, and its transcription factor, Nuclear factor erythroid 2-related factor 2 (Nrf2), were significantly induced by LRE treatment. Consequently, these results suggest that LRE ameliorates LPS-PG-induced inflammatory responses by attenuating TLR4/MyD88-mediated NF-κB, and activating NQO-1/Nrf2 antioxidant response element signaling pathways in HGF-1 cells. 적 요치주조직에 존재하는 주요한 세포의 한 형태인 인체 치은섬유아세포는 다양한 구강유해세균으로부터 염증이 유발되어지며, 그중 대표적으로 치주염 원인균인 P. gingivalis의 내독소인LPS-PG로부터 염증성 자극에 반응하여 다양한 염증매개 물질을 분비한다. 본 연구에서는 치주염을 일으키는 주요한 원인균중 하나인 P. gingivalis로 부터 분리한 LPS-PG를 이용하여 인체 치은섬유아세포주인 HGF-1 세포에 염증을 유도한 후 LRE 에 대한 항염증 및 항산화 효과를 분석하였다. 실험 결과, LRE 는 LPS-PG 유도에 따라 iNOS에 의한 NO 생성과 COX-2에 의한 PGE2와 같은 염증 매개 인자의 발현 및 생성 억제와 함께 염증성 싸이토카인(TNF-α, IL-1β및 IL-6)의 생성 또한 억제하였다. 신호전달계에서 염증성 전사인자의 발현 경로를 확인하기 위하여 TLR4/Myd88/NF-κB의 활성을 확인한 결과, LRE 처리에 따라 농도 의존적으로 억제되는 것을 확인하였다. 또한 산화 환원 효소로 항염증효과를 나타내는 것으로 알려진2상 효소중 하나인 NQO-1과 이의 전사인자인 Nrf2를 분석 한 결과 LRE 처리에 의해 효소의 활성이 높아지는 것을 확인할 수 있었다. 결론적으로 LRE는 TLR4/Myd88/NF-κB 신호전달 경로를 억제하고 NQO1/Nrf2 활성을 유도함으로써 HGF-1 세포에서LPS-PG에 의해 유도된 염증을 억제하는 것으로 사료되며, 향후 LRE는 식‧의약품 소재 개발에서 치주질환 개선의 가능성이 있는 후보물질이 될 수 있을 것으로 사료된다

Mesenchymal Stem Cells Modified with Stromal Cell-Derived Factor 1б Improve Cardiac Remodeling via Paracrine Activation of Hepatocyte Growth Factor in a Rat Model of Myocardial Infarction

Junming Tang,Jianing Wang,Linyun Guo,Xia Kong,Jianye Yang,Fei Zheng,Lei Zhang,Yongzhang Huang 한국분자세포생물학회 2010 Molecules and cells Vol.29 No.1

Mesenchymal stem cells (MSCs) are a promising source for cell-based treatment of myocardial infarction (MI), but existing strategies are restricted by low cell survival and engraftment. We examined whether SDF-1 transfection improve MSC viability and paracrine action in infarcted hearts. We found SDF-1-modified MSCs effectively ex-pressed SDF-1 for at least 21days after exposure to hy-poxia. The apoptosis of Ad-SDF-1-MSCs was 42% of that seen in Ad-EGFP-MSCs and 53% of untreated MSCs. In the infarcted hearts, the number of DAPI-labeling cells in the Ad-SDF-1-MSC group was 5-fold that in the Ad-EGFP-MSC group. Importantly, expression of antifibrotic factor, HGF, was detected in cultured MSCs, and HGF expression lev-els were higher in Ad-SDF-MSC-treated hearts, compared with Ad-EGFP-MSC or control hearts. Compared with the control group, Ad-SDF-MSC transplantation significantly decreased the expression of collagens I and III and matrix metalloproteinase 2 and 9, but heart function was im-proved in d-SDF-MSC-treated animals. In conclusion, SDF-1–modified MSCs enhanced the tolerance of engrafted MSCs to hypoxic injury in vitro and improved their viability in infarcted hearts, thus helping preserve the contractile function and attenuate left ventricle (LV) remodeling, and this may be at least partly mediated by enhanced paracrine signaling from MSCs via antifibrotic factors such as HGF.

Mesenchymal Stem Cells Modified with Stromal Cell-Derived Factor 1${\alpha}$ Improve Cardiac Remodeling via Paracrine Activation of Hepatocyte Growth Factor in a Rat Model of Myocardial Infarction

Tang, Junming,Wang, Jianing,Guo, Linyun,Kong, Xia,Yang, Jianye,Zheng, Fei,Zhang, Lei,Huang, Yongzhang Korean Society for Molecular and Cellular Biology 2010 Molecules and cells Vol.29 No.1

Mesenchymal stem cells (MSCs) are a promising source for cell-based treatment of myocardial infarction (MI), but existing strategies are restricted by low cell survival and engraftment. We examined whether SDF-1 transfection improve MSC viability and paracrine action in infarcted hearts. We found SDF-1-modified MSCs effectively expressed SDF-1 for at least 21days after exposure to hypoxia. The apoptosis of Ad-SDF-1-MSCs was 42% of that seen in Ad-EGFP-MSCs and 53% of untreated MSCs. In the infarcted hearts, the number of DAPI-labeling cells in the Ad-SDF-1-MSC group was 5-fold that in the Ad-EGFP-MSC group. Importantly, expression of antifibrotic factor, HGF, was detected in cultured MSCs, and HGF expression levels were higher in Ad-SDF-MSC-treated hearts, compared with Ad-EGFP-MSC or control hearts. Compared with the control group, Ad-SDF-MSC transplantation significantly decreased the expression of collagens I and III and matrix metalloproteinase 2 and 9, but heart function was improved in d-SDF-MSC-treated animals. In conclusion, SDF-1-modified MSCs enhanced the tolerance of engrafted MSCs to hypoxic injury in vitro and improved their viability in infarcted hearts, thus helping preserve the contractile function and attenuate left ventricle (LV) remodeling, and this may be at least partly mediated by enhanced paracrine signaling from MSCs via antifibrotic factors such as HGF.