Vitamin C activity of 2-O-alpha-D-glucopyranosyl-L-ascorbic acid in guinea pigs.

The vitamin C activity of 2-O-alpha-D-glucopyranosyl-L-ascorbic acid (AA-2G), which is one of chemically stable derivatives of L-ascorbic acid (AsA), in guinea pigs was investigated. Male guinea pigs were divided into 9 groups and fed AsA-deficient diet for 24 days with the following supplement: AA-2G- or AsA-supplemented groups were orally supplemented with 0.96, 1.92, 9.6 and 192 AA-2G mg/animal/day or equimolar amounts of AsA (0.5, 1, 5 and 100 mg/animal/day, respectively); AsA-deficient group received neither of them. The body weight gain, serum alkaline phosphatase activity, and the concentration of AsA and AA-2G in the liver, adrenals and urine of the guinea pigs were measured at the end of the experimental period. The AA-2G-supplemented guinea pigs showed similar body weight gain to the animals supplemented with equimolar amount of AsA. Serum alkaline phosphatase activity in both AA-2G- and AsA-supplemented groups was significantly higher than that of AsA-deficient group. But there was no significant difference between the groups supplemented with AA-2G and the equimolar amount of AsA. AA-2G-supplemented guinea pigs showed no apparent symptoms of scurvy. In AA-2G-supplemented groups, AA-2G was not detected in the liver, adrenals and urine, but AsA was found and the AsA concentration increased with increasing AA-2G dosage. The AsA concentration in the tissues of each AA-2G-supplemented groups was higher than that of AsA-deficient group, which was similar to that of the groups supplemented with equimolar amount of AsA. These results showed that AA-2G has the same vitamin C activity as AsA on a molar basis for the orally supplemented guinea pigs.     

Evaluation of ascorbic acid 2-O-alpha-glucoside as vitamin C source: mode of intestinal hydrolysis and absorption following oral administration.

Ascorbic acid 2-O-alpha-glucoside (AA-2G) has been reported to have antiscorbutic activity in guinea pigs. The present experiments examined the metabolic fate of AA-2G following ingestion. Oral administration of AA-2G (96 mg) to guinea pigs resulted in a remarkable increase of ascorbic acid in various tissues as well as plasma, but intact AA-2G was detected only in plasma, but intact AA-2G was detected only in plasma and urine in small amounts. The absorption efficiency of AA-2G and ascorbic acid was further determined by using everted gut sacs of rats. Ascorbic acid released from AA-2G on the mucosal side was effectively taken up across intestinal membranes into the serosal side, whereas AA-2G poorly permeated via a passive transport system. The hydrolysis of AA-2G on the mucosal surface of everted gut was completely inhibited by an alpha-glucosidase inhibitor and the hydrolytic activity of a crude membrane extract diminished to one-forth after immunoprecipitation with the antibody specific to maltase. From these results, it is concluded that ingested AA-2G serves as a vitamin C source through the hydrolysis by intestinal membrane-bound alpha-glucosidase, mainly maltase, and the subsequent absorption of released ascorbic acid.     

芦荟大黄素诱导硬皮病成纤维细胞凋亡的研究

目的探讨芦荟大黄素对系统性硬皮病患者皮肤成纤维细胞凋亡的影响。方法以不同浓度的芦荟大黄素处理体外培养的成纤维细胞,用TUNEL法和荧光显微镜观察法测定芦荟大黄素对成纤维细胞凋亡的诱导。结果芦荟大黄素对患者皮肤成纤维细胞的凋亡具有显著的诱导作用,该作用在一定范围内随着药物浓度的增高和作用时间的延长而增强。结论芦荟大黄素可诱导硬皮病患者皮肤成纤维细胞的凋亡,从而抑制其增殖,减少其胶原蛋白的合成。     

ERK1/2通路在AcSDKP抑制PDGF诱导的大鼠肺成纤维细胞增殖和胶原合成中的作用

目的 探讨细胞外信号调节激酶(ERK)1/2通路在N-乙酰基-丝氨酰-天门冬酰-赖氨酰-脯氨酸(AcSDKP)抑制血小板源性生长因子(PDGF)诱导的大鼠肺成纤维细胞增殖和胶原合成中的作用.方法 取新生Wistar大鼠20只,获得原代及传代培养的肺成纤维细胞.实验分为:(1)对照组(含体积分数为0.4%的胎牛血清的DMEM组);(2)PDGF组;(3)PD98059+PDGF组(25 μmol/L PD98059+10 ng/ml PDGF);(4)AcSDKP+PDGF组(1x10-8mol/L AcSDKP+10 ng/ml PDGF).采用噻唑蓝(MTT)法检测肺成纤维细胞的代谢活力,免疫细胞化学法、Western blot法检测Ⅰ、Ⅲ型胶原蛋白表达的改变;采用Western blot法检测ERK1/2及磷酸化-ERK1/2蛋白表达的改变.结果 与对照组相比,PDGF组大鼠肺成纤维细胞代谢活力增强,Ⅰ、Ⅲ型胶原蛋白表达增强,磷酸化-ERK1/2蛋白表达增高,差异均有统计学意义(P<0.05).AcSDKP+PDGF组细胞代谢活力明显低于PDGF组,免疫细胞化学法检测结果 显示,AcSDKP+PDGF组Ⅰ、Ⅲ型胶原蛋白表达分别为PDGF组的69.3%和67.2%.Western blot法检测结果 显示,AcSDKP+PDGF组细胞内Ⅰ、Ⅲ型胶原蛋白表达分别为PDGF组的92.4%和78.0%,磷酸化-ERK1/2蛋白表达为PDGF组的83.5%,差异均有统计学意义(P<0.05).结论 ERK1/2通路在AcSDKP抑制PDGF诱导的大鼠肺成纤维细胞增殖和胶原合成中发挥了重要作用.     

Recent advances in the study of the mechanisms of silica-induced pulmonary fibrosis

A review was made on the recent advances in the study on the pathogenesis of silica-induced pulmonary fibrosis. Alveolar macrophages which ingest silica particles liberate a fibrogenic factor, which stimulates the production of collagen of cultured fibroblasts. Silica deposited in the alveoli augments the demand of macrophages, the supply of which is maintained by monocytes recruited from the bone marrow. Attempts to demonstrate in vitro the presence of a fibrogenic factor in the supernatant of macrophages have been made in many laboratories, and an in vivo model utilizing diffusion chambers implanted in mice has been used by some investigators. A fibrogenic factor has been isolated and purified from the medium of silica-treated macrophages. Recent advances in immunological studies have demonstrated that silica stimulates macrophages to release monokines such as interleukin 1 (IL-1) and that IL-1 has chemical properties identical to the fibrogenic factor, which enhances the level of collagen production by modulating the proliferation of fibroblasts. Silica inhibits the suppressive effects of macrophages on fibroblasts. The increased protein synthesis in the fibroblasts is due partly to increase in mRNA. Collagen synthesis is stimulated not only by the fibrogenic factor released from silica-treated macrophages but also by the inhibition of macrophage ribonuclease activity. Information on the number of cells, collagen content and protease activity in the lung as well as in the bronchopulmonary lavage fluid has provided us a better understanding of the mechanisms involved in silica-induced pulmonary fibrosis.     

Zinc may increase bone formation through stimulating cell proliferation, alkaline phosphatase activity and collagen synthesis in osteoblastic MC3T3-E1 cells

Zinc is an essential trace element required for bone formation, however not much has been clarified yet for its role in osteoblast. We hypothesized that zinc would increase osteogenetic function in osteoblasts. To test this, we investigated whether zinc treatment enhances bone formation by stimulating osteoblast proliferation, bone marker protein alkaline phosphatase activity and collagen synthesis in osteoblastic MC3T3-E1 cells. MC3T3-E1 cells were cultured and treated with various concentrations of zinc (0, 1, 3, 15, 25 uM) along with a normal osteogenic medium (OSM) as control for 1, 5, 10 days. As measured by MTT assay for mitochondrial metabolic activity, cell proliferation was stimulated even at low zinc treatment (1-3 µM) compared to OSM, and it was stimulated in a zinc concentration-dependent manner during 5 and 10 days, with the most pronounced effect at 15 and 25 uM Zn. Cellular (synthesized) alkaline phosphatase (ALP) activity was increased in a zinc concentration-dependent manner, so did medium (secreted) ALP activity. Cellular collagen concentration was increased by zinc as time went by, therefore with the maximum zinc stimulatory effect in 10 days, and medium collagen concentration showed the same pattern even on 1 and 5 day. This zinc stimulatory effect of collagen synthesis was observed in cell matrix collagen staining. The study results imply that zinc can increase osteogenic effect by stimulating cell proliferation, ALP activity and collagen synthesis in osteoblastic cells.     

L-ascorbyl-2-monophosphate has equal antiscorbutic activity as L-ascorbic acid but L-ascorbyl-2-sulfate is inferior to L-ascorbic acid for channel catfish.

Channel catfish (Ictalurus punctatus) fingerlings (13 g average initial weight) were fed semipurified diets supplemented with 0, 0.06, 0.12, 0.24 and 0.72 mmol/kg (0, 11, 22, 44 or 132 mg/kg) of ascorbic acid molar equivalent supplied by either L-ascorbic acid, L-ascorbyl-2-monophosphate (Mg salt) (AAP), or L-ascorbyl-2-sulfate (K salt) (AAS). After 14 wk, weight gains were equal for all fish fed diets containing L-ascorbic acid or AAP; however, growth rates were less for fish fed AAS at all dietary levels and for fish fed the ascorbic acid-free diet (control). There were no gross signs of vitamin C deficiency in any of the fish fed L-ascorbic acid or AAP, whereas spinal deformities were found in the controls and in fish fed all but the highest concentration of AAS. The percentage of spinal deformities decreased as dietary levels of AAS increased. Reduced bone collagen content and histopathology in liver and gill tissues also indicated ascorbic acid deficiency in the controls and in fish fed all but the highest concentration of AAS. Limited histopathology was found in fish fed the lowest level of L-ascorbic acid but not in those fed the lowest level of AAP. Regression analysis of weight gain data showed that the vitamin activity of ascorbic acid from AAS was only 5.2% of that from L-ascorbic acid for growth. This study indicates that AAP has equimolar activity to L-ascorbic acid as a vitamin C source for channel catfish and that AAS has vitamin activity for this species but at a much lower level than the other compounds.     

Increase in the activity of alkaline phosphatase by L-ascorbic acid 2-phosphate in a human osteoblast cell line, HuO-3N1.

The activity of alkaline phosphatase (ALPase) was significantly enhanced in a human osteoblast cell line, HuO-3N1, when it was cultured in the presence of L-ascorbic acid 2-phosphate (AsA-P; a stable ascorbic acid derivative). With AsA-P in the culture, the level of ALPase activity increased approximately 3-fold without any effect on either the morphology or growth rate. This increase was dependent on the AsA-P concentration in the range of 0.2-2 mM and required at least 48 h incubation with AsA-P. The ALPase mRNA level, however, remained rather constant irrespective of the enzyme activity. Removal of AsA-P from the precultured medium decreased the stimulatory effect of ascorbic acid on the ALPase activity, indicating that the effect was reversible. Dexamethasone, an inducer for osteoblastic differentiation, enhanced the level of ALPase activity irreversibly, in parallel with the increase in the level of its mRNA. The enhancement of the ALPase activity by ascorbic acid in this cell line appeared to be independent of cell differentiation.     

Effects of soybean peptide and collagen peptide on collagen synthesis in normal human dermal fibroblasts

The collagen present in the dermis of the skin is a fibrous protein that fills the gaps between cells and helps maintain tissue flexibility. Effectively increasing the collagen present in the skin is an important goal for cosmetic research. Recent research has shown that soybean peptide (SP) has anti-fatigue activity, antioxidant activity, and the ability to increase type I collagen, while collagen peptide (CP) has the ability to enhance corneal moisture content and viscoelasticity, as well as to increase levels of hyaluronic acid synthesizing enzymes in human skin. Little documented research, however, has been conducted on collagen formation in relation to these peptides. Therefore, this research applied SP and CP with molecular weights primarily around 500 and preparations containing both SP and CP to normal human dermal fibroblasts together with magnesium ascorbyl phosphate (VC-PMg), and used real-time PCR to determine the gene expression of type I collagen (COL1A1), which contributes to collagen synthesis, and Smad7, which contribute to collagen breakdown. In addition, enzyme linked immuno sorbent assay (ELISA) was used to measure collagen content in the media. COL1A1 gene expression at 24 h after sample addition showed higher tendency in all samples and increased with time at 4, 8 and 24 h after addition. Smad7 gene expression was not substantially different at 4 h after addition. matrix metalloproteinase-1 gene expression was higher following SP addition, but was lower after the addition of CP and SP+CP. Medium collagen content was higher in all samples and increased with time at 8 h after addition. Collagen levels were higher when SP and CP were added together.     

石花菜乙酸乙酯提取物对增生性瘢痕成纤维细胞的影响

目的探讨石花菜乙酸乙酯提取物对人增生性瘢痕(HS)成纤维细胞增殖及合成功能的影响。方法组织植块法分离培养瘢痕组织成纤维细胞并鉴定,MTT法检测石花菜乙酸乙酯提取物(0,2.5,5,10 mg/ml)对细胞活性的影响;此外不同浓度的石花菜乙酸乙酯提取物(0,2.5,5,10 mg/ml)刺激成纤维细胞48 h后,ELISA法检测细胞培养上清中成纤维细胞Ⅰ、Ⅲ型胶原及转化生长因子β1(TGF-β1)分泌水平。结果石花菜乙酸乙酯提取物呈时间及剂量依赖性抑制成纤维细胞活性。此外,5～10 mg/ml石花菜提取物使HS成纤维细胞Ⅰ、Ⅲ型胶原生成及TGF-β1分泌显著少于对照组(p<0.05);2.5 mg/ml石花菜提取物虽不显著减少成纤维细胞的胶原合成,但与对照组相比,它可显著减少TGF-β1的分泌(p<0.05)。结论石花菜乙酸乙酯提取物能有效抑制增生性瘢痕成纤维细胞增殖,并抑制细胞胶原合成及生长因子的分泌,对增生性瘢痕可能具有治疗作用。     

煤尘刺激肺泡巨噬细胞对人胚肺成纤维细胞的作用

目的了解煤尘对肺成纤维细胞增殖的影响。方法采用不同浓度煤尘刺激兔肺泡巨噬细胞（ＰＡＭ）,观察其上清液对体外培养的人胚肺成纤维细胞（ＨＥＬＦ）的作用,测定ＨＥＬＦ的增殖程度、培养液上清中羟脯氨酸（Ｈｙｐ）含量,了解胶原合成能力。结果空白对照、ＰＡＭ、煤尘组和阳性对照组（二氧化硅）ＨＥＬＦ的增殖程度均数分别为０．２８５、０．３６８、０．３９８、０．７６８,Ｆ＝９３．９２,Ｐ＜０．０１。其中２００ｍｇ／Ｌ煤尘作用于ＰＡＭ后,其上清液对ＨＥＬＦ有显著刺激增殖作用（Ｐ＜０．０１）;１００、２００ｍｇ／Ｌ煤尘对ＨＥＬＦ的增殖率分别为９．７４％、１７．００％。１００ｍｇ／Ｌ组Ｈｙｐ含量为（３０．３９±０．２７）ｍｇ／Ｌ,明显高于ＰＡＭ对照组［（２７．５８±１．４６）ｍｇ／Ｌ］,Ｐ＜０．０５。结论一定浓度煤尘体外作用于ＰＡＭ,可刺激成纤维细胞增殖,同时伴有胶原合成能力的增加。     

Effects of soybean peptide and collagen peptide on collagen synthesis in normal human dermal fibroblasts

The collagen present in the dermis of the skin is a fibrous protein that fills the gaps between cells and helps maintain tissue flexibility. Effectively increasing the collagen present in the skin is an important goal for cosmetic research. Recent research has shown that soybean peptide (SP) has anti-fatigue activity, antioxidant activity, and the ability to increase type I collagen, while collagen peptide (CP) has the ability to enhance corneal moisture content and viscoelasticity, as well as to increase levels of hyaluronic acid synthesizing enzymes in human skin. Little documented research, however, has been conducted on collagen formation in relation to these peptides. Therefore, this research applied SP and CP with molecular weights primarily around 500 and preparations containing both SP and CP to normal human dermal fibroblasts together with magnesium ascorbyl phosphate (VC-PMg), and used real-time PCR to determine the gene expression of type I collagen (COL1A1), which contributes to collagen synthesis, and Smad7, which contribute to collagen breakdown. In addition, enzyme linked immuno sorbent assay (ELISA) was used to measure collagen content in the media. COL1A1 gene expression at 24?h after sample addition showed higher tendency in all samples and increased with time at 4, 8 and 24?h after addition. Smad7 gene expression was not substantially different at 4?h after addition. matrix metalloproteinase-1 gene expression was higher following SP addition, but was lower after the addition of CP and SP+CP. Medium collagen content was higher in all samples and increased with time at 8?h after addition. Collagen levels were higher when SP and CP were added together.     

成骨生长肽联合胰岛素样生长因子对成骨细胞增值的影响

目的探讨成骨生长肽（OGP）联合胰岛素样生长因子（IGF）对体外培养成骨细胞增值分化的影响。方法：将鼠成骨细胞分别与OGP（10-10mmol／L）、IGF-1（100ng／ml）、OGP（10-10mmol／L）＋IGF-1（100ng／ml）共同培养,于1、3、5d分别进行3H—TdR、3H-Tproline的掺入量与单独使用因子组、对照组相比,在各个时段都存在显著性差异（P〈0．01）;OGP联合IGF组对碱性磷酸酶合成量与对照组、OGP、IGF组相比有显著性差异（P〈0．01）。结论：OGP联合IGF可显著促进成骨细胞DNA、胶原蛋白、碱性磷酸酶的合成,对促进成骨细胞增值分化有协同作用。     

脂多糖对人正常皮肤成纤维细胞增殖周期及Ⅰ、Ⅲ型前胶原mRNA表达的影响

目的观察大肠杆菌脂多糖（lipopolysaccharide,LPS）刺激后,皮肤成纤维细胞增殖周期和Ⅰ、Ⅲ型前胶原mRNA表达变化规律。方法体外培养正常人皮肤成纤维细胞,加入不同浓度（0．005～1．0μg／m1）的大肠杆菌LPS（Ecoli055：B5）,于LPS加入后第7天细胞处于对数生长期时,采用流式细胞术观察细胞增殖周期;用逆转录-聚合酶链反应（RT-PCR）法测定成纤维细胞Ⅰ、Ⅲ型前胶原mRNA及胶原酶mRNA的表达。结果LPS刺激浓度在0．005～0．1μg／ml时,S期细胞比例随着刺激浓度增加而增高,且呈浓度依赖性;当LPS浓度为0．5μg／ml时,上述作用开始降低,但仍高于空白对照;当浓度达到1．0μg／ml时,s期细胞比例均低于空白对照;LPS刺激浓度在0．005～0．1μg／ml时,促进正常皮肤成纤维细胞Ⅰ、Ⅲ型前胶原mRNA表达,抑制胶原酶mRNA表达,且呈一定的剂量依赖性;当LPS刺激浓度为0．5μg／ml,上述作用下降;而当LPS刺激浓度到达1．0μg/ml时,抑制正常皮肤成纤维细胞Ⅰ、Ⅲ型前胶原mRNA表达,促进胶原酶mRNA表达。结论在一定浓度范围内LPS促进皮肤成纤维细胞增殖和胶原合成,而过高浓度LPS则呈现抑制效应。     

Enhanced oxidative stress by L-ascorbic acid within cells challenged by hydrogen peroxide

It has been amply documented that L-ascorbic acid added to the medium of a cell culture increases oxidative damage, and this effect of L-ascorbic acid has been ascribed to the generation of reactive oxygen intermediates in the medium during its auto-oxidation. We have here questioned whether such an effect is exerted inside the cell as well, and if so, what its mechanism is. To assess thiol oxidation in the cell, we manipulated CHO cells so that they could express bacterial alkaline phosphatase in the cytoplasm. Alkaline phosphatase activity, which requires the formation of intramolecular disulfide bridges, was shown to appear when the cells were exposed to H2O2. This H2O2-induced activity increased more than 1.5 fold when L-ascorbic acid had been loaded in the cells by incubation with L-ascorbic acid-2-O-phosphate. Similar enhancing effects were also observed by assessing oxidation of glutathione, formation of protein carbonyls, and generation of reactive oxygen intermediates. Interestingly, the effects by the L-ascorbic acid-2-O-phosphate treatment were totally suppressed by addition of the membrane-permeable chelator deferoxamine to the medium, indicating the involvement of iron ions. Because the apoprotein of conalbumin, which binds iron ions with a high affinity, had no effect and because the same deferoxamine effect was observed with the cells incubated in balanced salt solution with no metal salts added, it was concluded that L-ascorbic acid acts as a pro-oxidant within the cell suffering oxidative stress, and that this effect is elicited through increased redox-cycling of iron in combination with L-ascorbic acid.     

Lack of an association between cellular phospholipid triene: tetraene ratio and proliferation of human skin fibroblasts in culture

Two experiments were performed in an attempt to establish an association between cellular phospholipid triene:tetraene ratio and proliferation of human neonatal skin fibroblasts in culture. In Experiment 1, a low lipid culture medium was developed that caused an accumulation of (n-9) eicosatrienoic acid in the phospholipids of human fibroblasts. This culture medium, when supplemented with a mixture of mitogens, supported growth of human fibroblasts at a level equivalent to that found under conditions of maximal growth using serum supplementation (8% fetal bovine serum). The triene:tetraene ratio of fibroblast phospholipids under the two conditions was 1.88 vs. 0.03, suggesting that the growth of these cells was not adversely affected by a high (greater than 0.4) triene: tetraene ratio. In Experiment 2, cells were cultured in a low lipid, mitogen-supplemented medium with 16:1(n-7), 18:1(n-9), 18:2(n-6) or 20:4(n-6) added as the albumin complex. All the fatty acids permitted an equivalent maximal growth stimulation in the assay system, although having different effects on the phospholipid triene:tetraene ratio. The results suggest that there is a lack of an association between cellular phospholipid triene:tetraene ratio (range, 0.03 to 3.4) and proliferation of human fibroblasts in this culture system.     

MIF对大鼠动脉平滑肌细胞合成Ⅰ、Ⅲ型胶原蛋白的影响

目的了解巨噬细胞移动抑制因子（MIF）对平滑肌细胞合成胶原蛋白的影响。方法用巨噬细胞移动抑制因子组和对照组分别作用于体外培养大鼠动脉平滑肌细胞,通过Westernblot方法测定2组平滑肌细胞Ⅰ、Ⅲ型胶原蛋白含量。结果MIF组平滑肌细胞合成Ⅰ型胶原蛋白增多。但合成Ⅲ型胶原蛋白与对照组比较差异无统计学意义（P〉0．05）。结论MIF可以促进大鼠动脉平滑肌细胞合成Ⅰ型胶原蛋白。     

Effect of ascorbic acid and growth factors on collagen metabolism of flexor retinaculum cells from individuals with and without carpal tunnel syndrome

The effects of ascorbic acid and various growth factors on the proliferation rate and collagen metabolism were studied in cells from the flexor retinaculum of individuals with carpal tunnel syndrome (FR-CTS) and without carpal tunnel syndrome (FR control) and in human dermal fibroblasts. Ascorbic acid and four growth factors, including basic fibroblast growth factor, transforming growth factor, platelet-derived growth factor, and epidermal growth factors, were used. Ascorbic acid stimulates type I collagen production more in FR control than in FR-CTS. Growth factor treatment resulted in the following responses by the cells: (1) a higher mitogenic response than in the control cells; (2) a higher stimulation of type III collagen production and a lower stimulation of type I collagen production in CTS cells as compared with control cells; and (3) more alpha 2 (I) than alpha 1 (I) collagen production in CTS cells, unlike in control cells. We concluded that cells of the FR from individuals with CTS are physiologically altered.     

p38/ERK激酶调控TGF-β1诱导的HLF-02细胞Ⅰ型胶原表达及MMP-2活力

目的探讨转化生长因子β1(TGF-β1)/丝裂原活化蛋白激酶(MAPK)通路在调控人肺成纤维细胞Ⅰ型胶原表达、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)活力中的作用.方法 以人肺成纤维细胞HLF-02细胞系为研究对象,给予10 ng/ml的TGF-β1刺激不同时间;阻断实验以P38激酶特异性抑制剂(SB203580)、ERK激酶特异性抑制剂(PD98059)分别阻断p38通路和细胞外调控激酶(ERK)通路;采用逆转录聚合酶链反应(RT-PCR)检测细胞内Ⅰ型胶原mRNA的表达;运用Western印迹检测细胞上清液中Ⅰ型胶原蛋白表达;酶谱分析细胞上清液中MMP-2、MMP-9的活力.结果 (1)TGF-β1刺激HLF-02细胞,24、48、72 h组的Ⅰ型胶原mRNA表达水平分别为1.33±0.07、2.46±0.09、2.39±0.08;Ⅰ型胶原蛋白表达水平分别为114.89±8.95、208.16±6.75、211.46±8.05;MMP-2的活力分别为190.33±5.86、214.33±8.39、212.67±11.59.(2)SB203580对TGF-β1诱导的Ⅰ型胶原mRNA水平、蛋白水平及MMP-2活力的抑制率分别为51%、24%、20%.(3)PD98059对TGF-β1诱导的Ⅰ型胶原mRNA水平、蛋白水平以及MMP-2活力的抑制率分别为42%、13%、16%.结论 TGF-β1可促进HLF-02细胞Ⅰ型胶原的转录和蛋白表达,上调MMP-2活力;p38、ERK激酶通路在此过程中具有重要的调控作用.