牛肝菌提取液在细胞水平的抗衰老功效研究

通过水提取法分离提取得到牛肝菌提取液,生药质量浓度为60 g/L。采用XTT比色法检测牛肝菌提取液对表皮角质细胞的线粒体活性、观察细胞增殖并检测其对细胞DNA含量的影响;利用DCFH-DA等方法检测牛肝菌提取液对人皮肤成纤维细胞内活性氧(ROS)及抗氧化元件(ARE)的影响;利用免疫荧光法检测牛肝菌提取液对成纤维细胞Ⅰ型前胶原蛋白表达的影响。结果表明:牛肝菌提取液在终质量浓度为1.5 g/L时,较对照组可以显著提升角质细胞的线粒体活性及DNA的含量,能够明显促进细胞的增殖;牛肝菌提取液可以明显减少细胞内ROS的含量、提高细胞内ARE激活水平;同时牛肝菌提取液还可以促进Ⅰ型前胶原蛋白的表达。     

川芎嗪对肝星状细胞增殖及结缔组织生长因子表达的影响

目的探讨川芎嗪(TMP)对肝星状细胞HSC-T6增殖及结缔组织生长因子(CTGF)表达的影响。方法不同剂量的TMP作用于HSC-T6细胞不同时间后,MTT法检测细胞的增殖;ELISA法检测Ⅰ型、Ⅲ型胶原和透明质酸(HA)的合成;West-ern blot法测定CTGF的表达。结果100～1000mg/L的TMP作用于HSC-T6细胞后,细胞的增殖明显受到抑制,且呈剂量依赖性;Ⅰ型、Ⅲ型胶原和HA的合成减少,CTGF的表达也减少,二者下降程度呈正相关(r分别为0.727、0.643和0.769)。结论TMP能够有效抑制肝纤维化形成,其可能的机制为抑制HSC增殖并降低CTGF的表达,从而阻断细胞内基质的形成。     

当归净油抗衰老及抗炎祛痘功效的体外实验研究

通过人真皮成纤维细胞(HDF-a)模型,考察了当归净油对细胞分泌Ⅰ型胶原蛋白(Col-Ⅰ)的影响。结果表明,当归净油在最大安全质量浓度25 mg/L时,能有效增加HDF-a细胞分泌Col-Ⅰ达33.5%。利用痤疮丙酸杆菌刺激人急性单核细胞THP-1,建立了特异性痤疮炎症细胞模型,考察当归净油对细胞分泌炎症因子人白细胞介素1β(IL-1β)的影响。结果表明,当归净油最大安全质量浓度为12.5 mg/L,此时对痤疮丙酸杆菌诱导的THP-1细胞炎症因子IL-1β分泌抑制率高达94.51%,当质量浓度减半为6.25 mg/L时,抑制率为64.34%。当归净油作为一种芳香性中草药,有望成为抗衰老及抗炎祛痘的个人护理天然原料,在化妆品中得到进一步开发和利用。     

“太空人参酵母”在化妆品中潜在应用研究

通过体外细胞平台,从多角度对应用太空育种技术和现代发酵技术而获得的"太空人参酵母"提取物进行功效验证,为其在化妆品中的应用提供理论依据和科学指导。结果揭示"太空人参酵母"可以明显促进人原代角质细胞的增殖能力、克隆形成能力及划痕修复能力,并且可以促进基底层基底源生蛋白(Perlecan)的表达;此外"太空人参酵母"还有助于重离子辐照后的人原代成纤维细胞恢复细胞形态,恢复基底源生蛋白、Ⅰ型胶原蛋白和弹性蛋白表达的能力。以上结果表明,"太空人参酵母"具有提高细胞再生能力的活性,具有应用于促肌肤再生类化妆品的潜力。     

两组藏药组方的提取及功效研究

藏草药资源与化妆品的联合创新是将传承资源宝库与时尚产品结合,具有中药研究意义。本工作选取两组藏药组方(组方1为中麻黄、水柏枝、大籽蒿、高山柏、杜鹃、多脉猫乳、儿茶;组方2为白芥、藏木香、没药、多脉猫乳、紫草茸、红花)作为研究媒介,通过水与75%乙醇提取对比,对两个组方提取物中多糖、黄酮、酚类成分进行含量测定与质谱分析(UHPLC-MS-Q Exactive)。最终采用透明质酸酶抑制实验与角质细胞抗刺激试验分别对两个组方水提取物进行体外保湿与舒缓评价测试,结果表明两组方均有一定保湿作用,组方2具有一定的舒缓功效。     

南蛇藤提取物诱导SGC-7901胃癌细胞凋亡及其机制

目的观察南蛇藤乙酸乙酯提取物和正丁醇提取物在体外诱导SGC-7901胃癌细胞凋亡的作用,并初步探讨其分子机制。方法制备南蛇藤乙酸乙酯和正丁醇提取物,采用MTT法检测二者对SGC-7901胃癌细胞增殖的影响;应用FITC-AnnexinⅤ及碘化丙锭(PI)双标法,通过流式细胞仪(FCM)观察SGC-7901细胞的凋亡率及P53蛋白的表达。结果不同浓度的南蛇藤乙酸乙酯和正丁醇提取物(15、30、60、120μg/ml)对SGC-7901细胞的增殖均有一定的抑制作用,且呈剂量依赖性。两种南蛇藤提取物(60、120、240、480μg/ml)可剂量依赖性地诱导肿瘤细胞凋亡,且随着浓度的升高,SGC-7901细胞P53蛋白的表达也增加,浓度为240和480μg/ml的提取物组与对照组相比,差异有统计学意义。结论南蛇藤乙酸乙酯提取物和正丁醇提取物在体外可诱导SGC-7901细胞凋亡,上调细胞中P53基因的表达可能是其抗肿瘤作用的分子机制之一。     

3种茶提取物对MCF-7细胞增殖、凋亡的影响及其与蛋白激酶B的相关性

目的 探讨红茶、绿茶及普洱茶提取物对人乳腺癌细胞MCF-7增殖、凋亡的影响及其与蛋白激酶B(Protein ki-nase B,PKB)的相关性。方法用不同浓度的3种茶提取物处理MCF-7细胞不同时间,MTT法检测其对MCF-7细胞增殖的影响;Annexin V-FITC/PI双染流式细胞术检测其对细胞凋亡的影响;实时定量PCR检测细胞PKB基因mRNA的转录水平。结果 3种茶提取物对MCF-7细胞的增殖均具有明显的抑制作用,且呈时间、剂量依赖性;3种茶提取物处理的细胞早期凋亡率均高于未处理细胞,PKB基因mRNA的表达量明显降低。结论 3种茶提取物均含有可以抑制PKB转录活性的天然成分,并可诱导MCF-7细胞凋亡;其中绿茶诱导MCF-7细胞凋亡效果最明显。     

白梅花提取物抗氧化及美白功效评价

采用体外评价方法对白梅花提取物进行抗氧化及美白功效研究。通过测定白梅花提取物对超氧阴离子及羟自由基清除的半数抑制浓度(IC₅₀),以及在H₂O₂诱导人皮肤成纤维细胞株(HFF-1细胞)氧化损伤模型中,考察白梅花提取物对细胞形态学、细胞存活率、丙二醛(MDA)含量、活性氧自由基(ROS)含量、超氧化物歧化酶(SOD)活力、谷胱甘肽(GSH)活力、Ⅰ型胶原蛋白(ColⅠ)以及基质金属蛋白酶1(MMP-1)含量的影响,评价其抗氧化功效。采用酪氨酸酶活性抑制实验以及小鼠黑色素瘤细胞株(B16-F10细胞)黑色素含量测定实验评价白梅花提取物的美白功效。结果表明白梅花提取物具有自由基清除作用,且白梅花60%醇提物、白梅花90%醇提物可明显提高HFF-1细胞活力、SOD活力、GSH活力以及ColⅠ含量,下调ROS、MDA及MMP-1含量,并明显降低B16-F10细胞中的黑色素含量,表现出明显的抗氧化及美白功效。     

羟基富勒烯在抵抗皮肤光老化中发挥的功效

目的：制备可溶于水的羟基富勒烯,测试其抵抗光老化功效。方法：利用UVA照射构建人皮肤成纤维细胞光老化模型,通过检测ROS、胶原蛋白含量、细胞凋亡率等验证羟基富勒烯的抗光老化功效。结果显示,羟基富勒烯能够显著降低细胞ROS阳性率和细胞凋亡率,对Ⅰ型和Ⅲ型胶原蛋白具有很强的保护作用。结论：羟基富勒烯在抵抗光老化方面具有显著功效,可开发为化妆品的抗老化功效原料。     

金钗石斛复方在细胞水平的美容功效研究

通过水提取法得到金钗石斛复方提取液,生药质量浓度为0.5 g·mL-1。采用XTT比色法检测金钗石斛复方提取液对表皮角质细胞的线粒体活性、观察细胞增殖并检测其对ATP含量的影响;利用多巴染色等方法,检测金钗石斛复方提取液对黑素细胞酪氨酸酶活性的抑制和黑素生成的影响;利用免疫荧光法检测金钗石斛复方提取液对角质细胞保湿相关蛋白表达的影响。实验结果表明,金钗石斛复方提取液在终质量浓度为2.5 g·L-1时,较对照组可以显著提升表皮细胞的线粒体活性和ATP含量,能够明显促进细胞的增殖。金钗石斛复方提取液在不影响共培养细胞状态的前提下,较对照组可以明显降低黑素细胞的黑色素含量并抑制酪氨酸酶活性。同时金钗石斛复方还可以促进角质细胞水通道蛋白AQP3、紧密连接蛋白ZO-1及Claudin-1的表达。     

Lidocaine Inhibited Tendon Cell Proliferation and Extracellular Matrix Production by Down Regulation of Cyclin A,CDK2, Type I and Type III Collagen Expression

Lidocaine injection is a common treatment for tendon injuries. However, the evidence suggests that lidocaine is toxic to tendon cells. This study investigated the effects of lidocaine on cultured tendon cells, focusing on the molecular mechanisms underlying cell proliferation and extracellular matrix (ECM) production. Tendon cells cultured from rat Achilles tendons were treated with 0.5, 1.0, or 1.5 mg/mL lidocaine for 24 h. Cell proliferation was evaluated by Cell Counting Kit 8 (CCK-8) assay and bromodeoxyuridine (BrdU) assay. Cell apoptosis was assessed by Annexin V and propidium iodide (PI) stain. Cell cycle progression and cell mitosis were assessed through flow cytometry and immunofluorescence staining, respectively. The expression of cyclin E, cyclin A, cyclin-dependent kinase 2 (CDK2), p21, p27, p53, matrix metalloproteinases-2 (MMP-2), matrix metalloproteinases-9 (MMP-9), type I collagen, and type III collagen were examined through Western blotting, and the enzymatic activity of MMP-9 was determined through gelatin zymography. Lidocaine reduced cell proliferation and reduced G1/S transition and cell mitosis. Lidocaine did not have a significant negative effect on cell apoptosis. Lidocaine significantly inhibited cyclin A and CDK2 expression but promoted p21, p27, and p53 expression. Furthermore, the expression of MMP-2 and MMP-9 increased, whereas that of type I and type III collagen decreased. Lidocaine also increased the enzymatic activity of MMP-9. Our findings support the premise that lidocaine inhibits tendon cell proliferation by changing the expression of cell-cycle-related proteins and reduces ECM production by altering levels of MMPs and collagens.     

Asthmatic Eosinophils Alter the Gene Expression of Extracellular Matrix Proteins in Airway Smooth Muscle Cells and Pulmonary Fibroblasts

The impaired production of extracellular matrix (ECM) proteins by airway smooth muscle cells (ASMC) and pulmonary fibroblasts (PF) is a part of airway remodeling in asthma. This process might be influenced by eosinophils that migrate to the airway and abundantly secrete various cytokines, including TGF-β. We aimed to investigate the effect of asthmatic eosinophils on the gene expression of ECM proteins in ASMC and PF. A total of 34 study subjects were recruited: 14 with allergic asthma (AA), 9 with severe non-allergic eosinophilic asthma (SNEA), and 11 healthy subjects (HS). All AA patients underwent bronchial allergen challenge with D. pteronyssinus. The peripheral blood eosinophils were isolated using high-density centrifugation and magnetic separation. The individual cell cultures were made using hTERT ASMC and MRC-5 cell lines and the subjects’ eosinophils. The gene expression of ECM and the TGF-β signaling pathway was analyzed using qRT-PCR. We found that asthmatic eosinophils significantly promoted collagen I, fibronectin, versican, tenascin C, decorin, vitronectin, periostin, vimentin, MMP-9, ADAM33, TIMP-1, and TIMP-2 gene expression in ASMC and collagen I, collagen III, fibronectin, elastin, decorin, MMP-2, and TIMP-2 gene expression in PF compared with the HS eosinophil effect. The asthmatic eosinophils significantly increased the gene expression of several canonical and non-canonical TGF-β signaling pathway components in ASMC and PF compared with the HS eosinophil effect. The allergen-activated AA and SNEA eosinophils had a greater effect on these changes. In conclusion, asthmatic eosinophils, especially SNEA and allergen-activated eosinophils, imbalanced the gene expression of ECM proteins and their degradation-regulating proteins. These changes were associated with increased gene expression of TGF-β signaling pathway molecules in ASMC and PF.     

Extracellular Matrix Biomarkers in Colorectal Cancer

The cellular microenvironment composition and changes therein play an extremely important role in cancer development. Changes in the extracellular matrix (ECM), which constitutes a majority of the tumor stroma, significantly contribute to the development of the tumor microenvironment. These alterations within the ECM and formation of the tumor microenvironment ultimately lead to tumor development, invasion, and metastasis. The ECM is composed of various molecules such as collagen, elastin, laminin, fibronectin, and the MMPs that cleave these protein fibers and play a central role in tissue remodeling. When healthy cells undergo an insult like DNA damage and become cancerous, if the ECM does not support these neoplastic cells, further development, invasion, and metastasis fail to occur. Therefore, ECM-related cancer research is indispensable, and ECM components can be useful biomarkers as well as therapeutic targets. Colorectal cancer specifically, is also affected by the ECM and many studies have been conducted to unravel the complex association between the two. Here we summarize the importance of several ECM components in colorectal cancer as well as their potential roles as biomarkers.     

血脂康对新生大鼠心肌成纤维细胞增殖及胶原合成的影响

目的观察血脂康对血管紧张素Ⅱ(AngⅡ)诱导的新生大鼠心肌成纤维细胞(CFs)增殖及胶原合成的影响,并探讨可能的分子机制。方法采用胰酶消化法分离培养Sprague-Dawley大鼠心肌成纤维细胞,建立AngⅡ诱导CFs增殖的模型,以MTT比色法和流式细胞仪检测细胞周期分析法观察血脂康对CFs数目和细胞周期的影响。AngⅡ及不同浓度血脂康作用48 h后,用天狼星红染色法检测培养上清中胶原的含量;ELISA法检测细胞培养上清液中TGF-β1蛋白表达;RT-PCR法检测胶原和TGF-β1 mRNA表达。结果AngⅡ对CFs增殖有明显促进作用,血脂康可明显抑制AngⅡ诱导的CFs增殖,且呈剂量依赖性;血脂康可增加G0/G1期细胞百分率,降低S、G2/M期细胞百分率,降低胶原含量、TGF-β1蛋白及胶原和TGF-β1 mRNA的表达。结论血脂康能抑制AngⅡ诱导的CFs增殖和胶原的产生,其作用可能是通过抑制TGF-β1表达实现的。     

Collagen I Modifies Connexin-43 Hemichannel Activity via Integrin α2β1 Binding in TGFβ1-Evoked Renal Tubular Epithelial Cells

Chronic Kidney Disease (CKD) is associated with sustained inflammation and progressive fibrosis, changes that have been linked to altered connexin hemichannel-mediated release of adenosine triphosphate (ATP). Kidney fibrosis develops in response to increased deposition of extracellular matrix (ECM), and up-regulation of collagen I is an early marker of renal disease. With ECM remodeling known to promote a loss of epithelial stability, in the current study we used a clonal human kidney (HK2) model of proximal tubular epithelial cells to determine if collagen I modulates changes in cell function, via connexin-43 (Cx43) hemichannel ATP release. HK2 cells were cultured on collagen I and treated with the beta 1 isoform of the pro-fibrotic cytokine transforming growth factor (TGFβ1) ± the Cx43 mimetic Peptide 5 and/or an anti-integrin α2β1 neutralizing antibody. Phase microscopy and immunocytochemistry observed changes in cell morphology and cytoskeletal reorganization, whilst immunoblotting and ELISA identified changes in protein expression and secretion. Carboxyfluorescein dye uptake and biosensing measured hemichannel activity and ATP release. A Cytoselect extracellular matrix adhesion assay assessed changes in cell-substrate interactions. Collagen I and TGFβ1 synergistically evoked increased hemichannel activity and ATP release. This was paralleled by changes to markers of tubular injury, partly mediated by integrin α2β1/integrin-like kinase signaling. The co-incubation of the hemichannel blocker Peptide 5, reduced collagen I/TGFβ1 induced alterations and inhibited a positive feedforward loop between Cx43/ATP release/collagen I. This study highlights a role for collagen I in regulating connexin-mediated hemichannel activity through integrin α2β1 signaling, ahead of establishing Peptide 5 as a potential intervention.     

The Beneficial Effect of Rosmarinic Acid on Benzophenone-3-Induced Alterations in Human Skin Fibroblasts

Benzophenone-3 (BP-3) is one of the most widely used chemical sunscreens. The results of many in vitro and in vivo tests confirm its high percutaneous penetration and systemic absorption, which question the safety of its wide use. The aim of our research was to assess the effect of this compound on components of the skin extracellular matrix, and to investigate whether rosmarinic acid (RA) could reduce BP-3-induced changes in human skin fibroblasts. BP-3 used at concentrations of 0.1–100 µM caused a number of unfavorable changes in the level of type I collagen, decorin, sulfated glycosaminoglycans, hyaluronic acid, elastin, and expression or activity of matrix metalloproteinases (MMP-1, MMP-2), elastase and hyaluronidase. Moreover, the intracellular retention of collagen was accompanied by changes in the expression of proteins modifying and controlling the synthesis and secretion of this protein. Most importantly, RA at a concentration of 100 µM significantly reduced or completely abolished the adverse effects of BP-3. Based on these findings, it can be concluded that this polyphenol may provide effective protection against BP-3-induced disturbances in skin cells, which may have important clinical implications.     

金属蛋白酶在皮肤构造与老化中所扮的角色(英文)

皮肤细胞间质体 (ECM)是由表皮与真皮产生的各种蛋白构成。破坏ECM的金属蛋白酶(MMP)是引起皮肤老化的重要原因。年龄的增长、环境的影响使皮肤逐渐老化。从分子与细胞层面的角度出发 ,着重分析了ECM与MMP对此所起的根本作用。年龄与紫外线影响会激增MMP2 (MMP的一种 )的活性 ,降低内在TIMP (抑制MMP2的酶 )的作用 ,使胶原蛋白纤维逐渐衰败 ,失去健康的平衡。MMP2激增的活性破坏在表皮与真皮间起支托作用的胶原蛋白 7型 ,可导致皱纹 ;MMP2还破坏胶原蛋白 1型 ,使ECM缺损、萎缩 ,导致微细血管扩张 ,可表现为表面网状血管与黑眼圈现象