目的观察加减射干麻黄汤对哮喘大鼠气道重塑及肺组织增殖细胞核抗原(PCNA)与细胞外调节蛋白激酶(ERK)的影响并探讨其机制。方法 60只Wistar大鼠随机分为模型对照组、空白对照组、中药治疗组、地塞米松组。使用卵蛋白致敏建立哮喘大鼠模...目的观察加减射干麻黄汤对哮喘大鼠气道重塑及肺组织增殖细胞核抗原(PCNA)与细胞外调节蛋白激酶(ERK)的影响并探讨其机制。方法 60只Wistar大鼠随机分为模型对照组、空白对照组、中药治疗组、地塞米松组。使用卵蛋白致敏建立哮喘大鼠模型,中药治疗组予加减射干麻黄汤0.5 m L灌胃,地塞米松组用地塞米松(0.65 mg/kg)0.26 m L腹腔注射。利用图像分析软件测量大鼠支气管内管壁面积、平滑肌面积、基底膜周长,通过免疫组化法检测大鼠肺组织PCNA和ERK的表达,原位杂交法检测大鼠肺组织PCNA和ERK的m RNA的表达,免疫印迹法检测大鼠肺组织PCNA和ERK的表达。结果 1)中药治疗组哮喘大鼠的症状明显减轻。2)中药治疗组大鼠肺组织支气管上皮细胞的炎症反应减轻,支气管壁及管腔内的炎症细胞也明显减少(P<0.05)。3)中药治疗组WAm/Pbm及WAi/Pbm明显降低(P<0.05),与地塞米松组比较无显著差异(P>0.05)。4)中药治疗组与地塞米松组PCNA m RNA和ERK m RNA表达较模型对照组明显减少(P<0.05)。5)中药治疗组PCNA m RNA和ERK m RNA表达较模型对照组明显减少,且与地塞米松组相当(P>0.05)。6)中药治疗组PCNA、p-ERK、ERK蛋白表达较哮喘模型组明显降低(P<0.05);与地塞米松组比较明显降低(P<0.05)。结论加减射干麻黄汤能改善哮喘大鼠气道重塑,其机制可能是通过降低PCNA的水平,抑制气道平滑肌增殖及ERK、p-ERK、PCNA蛋白表达水平,从而达到改善哮喘气道重塑的目的。展开更多
AIM： To identify the protein expression differences related to the CagA-induced ERK pathway activation in AGS cells. METHODS： Human AGS cells transfected with cagA and blank vector were treated with specific mitogen...AIM： To identify the protein expression differences related to the CagA-induced ERK pathway activation in AGS cells. METHODS： Human AGS cells transfected with cagA and blank vector were treated with specific mitogenactivated protein kinase kinase （MEK） inhibitor. Total cell proteins were combined by strong anion exchange （SAX2） and weak cation exchange （CM10） ProteinChip arrays and analyzed using surface-enhanced laser desorption/ ionization time-of-flight mass spectrometry （SELDI-TOF- MS） proteomics technology. Protein expression profiles were compared with those of inhibitor-untreated cagA transfectants. SwissProt/TrEMBL database searching for differentially expressed proteins was carried out using the TagIdent tool with the pI and mass information. RESULTS： When a total of 16 proteins that showed expression differences in inhibitor-untreated cagA transfectants were compared with vector transfectants, three proteins with m/z 4229, 8162 and 9084 were found to have no expression differences after treatment with MEK inhibitor, while the other 13 maintained the same expression differences after inhibitor treatment. Seven pieces of meaningful matching information for the three proteins were obtained from database searching. CONCLUSION： Biomarkers with m/z 4229, 8162 and 9084 are ERKI/2 phosphorylation dependent, andtherefore are the downstream molecules of ERK1/2 in the ERK/MAPK signaling pathway. The three biomarkers may be important cancer-associated proteins according to SwissProt/TrEMBL database information.展开更多
Accumulating evidence indicates that endostatin inhibits fibrosis. However, the mechanism is yet to be clarified. The aim of this study is to evaluate the effect of endostatin on platelet-derived growth factor-BB （PD...Accumulating evidence indicates that endostatin inhibits fibrosis. However, the mechanism is yet to be clarified. The aim of this study is to evaluate the effect of endostatin on platelet-derived growth factor-BB （PDGF-BB）- or transforming growth factor β1 （TGF-β1）-induced fibrosis in cultured human skin fibroblasts, and to further examine the molecular mechanisms involved. Human dermal flbroblasts were cultured in Dulbecco＇s modified Eagle＇s medium （DMEM） and serum-starved for 48 h before treatment. Cells were grouped as follows： ＂PDGF-BB＂, ＂PDGF-BB＋ endostatin＂, ＂TGF-β1＂, ＂TGF-β1＋endostatin＂, ＂endostatin＂, and ＂blank control＂. The fibroblasts were stimulated with either TGF-β1 or PDGF-BB for 72 h in order to set up the fibrosis model in vitro. The cells were co-cultured with either TGF-β1 or PDGF-BB and endostatin and were used to check the inhibiting effect of endostatin. A blank control group and an endostatin group were used as negative control groups. The biomarkers of fibrosis, including the expression of collagen I, hydrroxyproline, and α-smooth muscle actin （a-SMA）, were evaluated using an enzyme-linked immune- sorbent assay （ELISA） and Western blot. The expression of phosphorylated PDGF receptor β （p-PDGFRβ）, PDGFRβ, phosphorylated extracellular signal-regulated kinase （p-ERK）, and ERK was detected using Western blot and im- munofiuorescent staining was used to explore the mechanisms. Both PDGF-BB and TGF-β1 significantly up-regulated the expression of collagen I, hydroxyproline, and a-SMA. Endostatin significantly attenuated both the PDGF-BB- and TGF-β1-induced over-expression of collagen I, hydroxyproline, and a-SMA. PDGF-BB and TGF-β1 both promoted the expression of PDGFR, ERK, and p-ERK. Endostatin inhibited the expression of PDGFR and p-ERK but did not affect the expression of total ERK. Endostatin inhibited hypertrophic scar by modulating the PDGFRI3/ERK pathway. En- dostatin could be a promising multi-target drug in future展开更多
The differentiation of periodontal ligament(PDL)progenitor cells is important for maintaining the homeostasis of PDL tissue and alveolar bone.Vitamin C(VC),a water-soluble nutrient that cannot be biosynthesized by hum...The differentiation of periodontal ligament(PDL)progenitor cells is important for maintaining the homeostasis of PDL tissue and alveolar bone.Vitamin C(VC),a water-soluble nutrient that cannot be biosynthesized by humans,is vital for mesenchymal stem cells differentiation and plays an important role in bone remodeling.Therefore,the objective of this study was to determine the function and mechanism of VC in PDL progenitor cells osteogenic differentiation at the molecular level.We demonstrated that VC could induce the osteogenic differentiation and maturation of PDL progenitor cell without other osteogenic agents.During the process,VC preferentially activated ERK1/2 but did not affect JNK or p38.Co-treatment with ERK inhibitor effectively decreased the Vitamin C-induced expression of Runx2.ERK inhibitor also abrogated Vitamin C-induced the minimized nodules formation.PELP1,a nuclear receptor co-regulator,was up-regulated under VC treatment.PELP1 knockdown inhibited ERK phosphorylation.The overexpression of PELP1 had a positive relationship with Runx2 expression.Taken together,we could make a conclude that VC induces the osteogenic differentiation of PDL progenitor cells via PELP1-ERK axis.Our fi nding implies that VC may have a potential in the regeneration medicine and application to periodontitis treatment.展开更多
文摘目的观察加减射干麻黄汤对哮喘大鼠气道重塑及肺组织增殖细胞核抗原(PCNA)与细胞外调节蛋白激酶(ERK)的影响并探讨其机制。方法 60只Wistar大鼠随机分为模型对照组、空白对照组、中药治疗组、地塞米松组。使用卵蛋白致敏建立哮喘大鼠模型,中药治疗组予加减射干麻黄汤0.5 m L灌胃,地塞米松组用地塞米松(0.65 mg/kg)0.26 m L腹腔注射。利用图像分析软件测量大鼠支气管内管壁面积、平滑肌面积、基底膜周长,通过免疫组化法检测大鼠肺组织PCNA和ERK的表达,原位杂交法检测大鼠肺组织PCNA和ERK的m RNA的表达,免疫印迹法检测大鼠肺组织PCNA和ERK的表达。结果 1)中药治疗组哮喘大鼠的症状明显减轻。2)中药治疗组大鼠肺组织支气管上皮细胞的炎症反应减轻,支气管壁及管腔内的炎症细胞也明显减少(P<0.05)。3)中药治疗组WAm/Pbm及WAi/Pbm明显降低(P<0.05),与地塞米松组比较无显著差异(P>0.05)。4)中药治疗组与地塞米松组PCNA m RNA和ERK m RNA表达较模型对照组明显减少(P<0.05)。5)中药治疗组PCNA m RNA和ERK m RNA表达较模型对照组明显减少,且与地塞米松组相当(P>0.05)。6)中药治疗组PCNA、p-ERK、ERK蛋白表达较哮喘模型组明显降低(P<0.05);与地塞米松组比较明显降低(P<0.05)。结论加减射干麻黄汤能改善哮喘大鼠气道重塑,其机制可能是通过降低PCNA的水平,抑制气道平滑肌增殖及ERK、p-ERK、PCNA蛋白表达水平,从而达到改善哮喘气道重塑的目的。
基金National High Technology 2006AA02Z341 & NSFC 30430730
文摘AIM： To identify the protein expression differences related to the CagA-induced ERK pathway activation in AGS cells. METHODS： Human AGS cells transfected with cagA and blank vector were treated with specific mitogenactivated protein kinase kinase （MEK） inhibitor. Total cell proteins were combined by strong anion exchange （SAX2） and weak cation exchange （CM10） ProteinChip arrays and analyzed using surface-enhanced laser desorption/ ionization time-of-flight mass spectrometry （SELDI-TOF- MS） proteomics technology. Protein expression profiles were compared with those of inhibitor-untreated cagA transfectants. SwissProt/TrEMBL database searching for differentially expressed proteins was carried out using the TagIdent tool with the pI and mass information. RESULTS： When a total of 16 proteins that showed expression differences in inhibitor-untreated cagA transfectants were compared with vector transfectants, three proteins with m/z 4229, 8162 and 9084 were found to have no expression differences after treatment with MEK inhibitor, while the other 13 maintained the same expression differences after inhibitor treatment. Seven pieces of meaningful matching information for the three proteins were obtained from database searching. CONCLUSION： Biomarkers with m/z 4229, 8162 and 9084 are ERKI/2 phosphorylation dependent, andtherefore are the downstream molecules of ERK1/2 in the ERK/MAPK signaling pathway. The three biomarkers may be important cancer-associated proteins according to SwissProt/TrEMBL database information.
基金Project supported by the Zhejiang Provincial Natural Science Foundation of China(No.LY15H150004)the Teaching Department of the Zhejiang Province(No.Y201330073),China
文摘Accumulating evidence indicates that endostatin inhibits fibrosis. However, the mechanism is yet to be clarified. The aim of this study is to evaluate the effect of endostatin on platelet-derived growth factor-BB （PDGF-BB）- or transforming growth factor β1 （TGF-β1）-induced fibrosis in cultured human skin fibroblasts, and to further examine the molecular mechanisms involved. Human dermal flbroblasts were cultured in Dulbecco＇s modified Eagle＇s medium （DMEM） and serum-starved for 48 h before treatment. Cells were grouped as follows： ＂PDGF-BB＂, ＂PDGF-BB＋ endostatin＂, ＂TGF-β1＂, ＂TGF-β1＋endostatin＂, ＂endostatin＂, and ＂blank control＂. The fibroblasts were stimulated with either TGF-β1 or PDGF-BB for 72 h in order to set up the fibrosis model in vitro. The cells were co-cultured with either TGF-β1 or PDGF-BB and endostatin and were used to check the inhibiting effect of endostatin. A blank control group and an endostatin group were used as negative control groups. The biomarkers of fibrosis, including the expression of collagen I, hydrroxyproline, and α-smooth muscle actin （a-SMA）, were evaluated using an enzyme-linked immune- sorbent assay （ELISA） and Western blot. The expression of phosphorylated PDGF receptor β （p-PDGFRβ）, PDGFRβ, phosphorylated extracellular signal-regulated kinase （p-ERK）, and ERK was detected using Western blot and im- munofiuorescent staining was used to explore the mechanisms. Both PDGF-BB and TGF-β1 significantly up-regulated the expression of collagen I, hydroxyproline, and a-SMA. Endostatin significantly attenuated both the PDGF-BB- and TGF-β1-induced over-expression of collagen I, hydroxyproline, and a-SMA. PDGF-BB and TGF-β1 both promoted the expression of PDGFR, ERK, and p-ERK. Endostatin inhibited the expression of PDGFR and p-ERK but did not affect the expression of total ERK. Endostatin inhibited hypertrophic scar by modulating the PDGFRI3/ERK pathway. En- dostatin could be a promising multi-target drug in future
基金This work is supported by grants:the National Basic Research Program(973 Program)(No.2011CB707705)National Natural Science Foundation of China(Grant No.11202229).
文摘The differentiation of periodontal ligament(PDL)progenitor cells is important for maintaining the homeostasis of PDL tissue and alveolar bone.Vitamin C(VC),a water-soluble nutrient that cannot be biosynthesized by humans,is vital for mesenchymal stem cells differentiation and plays an important role in bone remodeling.Therefore,the objective of this study was to determine the function and mechanism of VC in PDL progenitor cells osteogenic differentiation at the molecular level.We demonstrated that VC could induce the osteogenic differentiation and maturation of PDL progenitor cell without other osteogenic agents.During the process,VC preferentially activated ERK1/2 but did not affect JNK or p38.Co-treatment with ERK inhibitor effectively decreased the Vitamin C-induced expression of Runx2.ERK inhibitor also abrogated Vitamin C-induced the minimized nodules formation.PELP1,a nuclear receptor co-regulator,was up-regulated under VC treatment.PELP1 knockdown inhibited ERK phosphorylation.The overexpression of PELP1 had a positive relationship with Runx2 expression.Taken together,we could make a conclude that VC induces the osteogenic differentiation of PDL progenitor cells via PELP1-ERK axis.Our fi nding implies that VC may have a potential in the regeneration medicine and application to periodontitis treatment.
