Effect of ALA-PDT on the expressions of MMP-9, MMP-13 and TIMP-1 of hypertrophic scar model in rabbit ears

Objective: To investigate the effect of 5-aminolevulinic(ALA)-photodynamic therapy(PDT) on the expressions of MMP-9, MMP-13 and TIMP-1 of hypertrophic scar model in rabbit ears, and analyze the possible therapeutic mechanisms of ALA-PDT treatment to hypertrophic scars of rabbit ears. Methods: The experimental animals were randomly divided into normal control, negative control, high concentration of ALA-PDT, low concentration of ALA-PDT and PDT groups. The latter three groups received ALA-PDT treatment or PDT treatment once a week for 3 weeks. The specimens of the rabbits were collected respectively 1, 2 and 3 months after treatment to be used for RT-PCR and Western-blot test. Results: 1, 2 and 3 months after PDT treatment, the expressions of MMP-9 and MMP-13(including mRNA and protein) in hypertrophic scar tissues of three treatment groups were significantly higher than those of the negative control group(P<0.01), and the expression of TIMP-1 mRNA and protein of three treatment groups were significantly lower than that of the negative control group(P<0.01). There were also significant differences between high-concentration ALA-PDT treatment group and the low one(P<0.05). Conclusion: ALA-PDT is effective in treating hypertrophic scars of rabbit ears, and its possible therapeutic mechanisms are that ALA-PDT treatment generates oxidation activation effect to activate the activity of MMPs and induces the photoaging of fibroblasts of hypertrophic scar tissues of rabbit ears to inhibit the activity of TIMPs, which causes the up-regulation of MMPs and the down-regulation of TIMPs. Because of this, the degradation of collagen and ECM is accelerated and the formation of scars is suppressed.

Expression of MMP-9 and -13 on the Pressure Side under Orthodontic Loading

Purpose: The purpose of this study was to investigate the expression of MMP-9 and MMP-13 on compression side of tooth movement. Material and Methods: Twenty male Wistar rats were used for this experiment. Maxillary right first molar was moved orthodontically with a constant force of 20 g and on the side of the maxillary left first molar was used as control group. On the 4th, 10th and 17th days after experiment, 5 rats were euthanized, respectively. Histologic process was processed for immunohistochemical staining of MMP-9 and -13. Results: MMP-9 and -13 expressions were upregulated at day 4 after tooth movement. The expression of MMP-9 was not observed at days 7, 10 and 14, while the expression of MMP-13 was greatly increased at days 7 and 10. Conclusion: This study suggests that different characteristics of MMPs expression may contribute to the remodeling process of collagen fibers in the PDL during tooth movement.

Expression of MMP-2 and MMP-13 in Bullous Pemphigoid

Objective:To investigate the expression and significance of matrix metalloproteinase-2(MMP-2)and matrix metalloproteinase-13(MMP-13)in bullous pemphigoid(BP)skin lesions.Methods:Immunohistochemical SP method was used to detect the expression of MMP-2 and MMP-13 in 32 BP skin lesions,and compared with 15 normal skin tissues.Results:The expression of MMP-2 in the case group was significantly increased(38.56±10.06)compared to the normal control group(21.20±5.98);the expression of MMP-13 in the case group was significantly augmented(18.62±5.90)compared to the normal control group(11.47±8.484).The expressions of MMP-2 and MMP-13 in the skin lesions of patients with bullous pemphigoid were statistically different from those of normal people(both P<0.05).Compared with the expression of MMP-2 and MMP-13 in bullous pemphigoid,the expression of MMP-2 and MMP-13 was moderately correlated(correlation coefficient was 0.523).Conclusion:The expression of MMP-2 and MMP-13 is significantly increased in bullous pemphigoid skin lesions,suggesting that they may play an important role in the pathogenesis of BP.There is a certain correlation between the expression of MMP-2 and MMP-13,suggesting that the high expression of MMP-13 may play a role in the mechanism that further leads to the high expression of MMP-2.

Characterization of ST13 Protein Expression in Human Colorectal Cancer Tissues

Objective: To characterize the expression of ST13 protein in human tissuesfor investigation of the function of colorectal cancer related gene ST13. Methods: ST13 ORF wascloned and over-expressed in E.coli. The recombinant ST13 protein was purified by affinitychromatography. ST13 monoclonal antibodies were generated and affinity purified with the recombinantprotein. Immunoblot and immunohistochemical staining were employed to analyze ST13 proteinexpression in human tissues. Results: The expression and purification of the recombinant ST13protein were confirmed by SDS-PAGE. The protein yield reached about 2.5 mg/L of induced bacterialculture with a purity of 91.3%. Three strains of hybridoma were obtained with antibody titers from10~4 to 10~5 in ascites fluids and with high specificity for ST13 protein. Immunoblot showed thatthe apparent Mr of ST13 protein in SW480 cells and human tissues estimated by SDS-PAGE mobility wasapproximately 50 000, which was about 10 000 larger than the 41 324 calculated, but theglycosylation of the protein was excluded. Computer modeling revealed the protein to be ahydrophilic molecule. Immunohistochemical staining showed that ST13 protein was evenly distributedin cytoplasm and expressed in colon, stomach, liver, and other epithelial cells. Differences in thestaining intensity of the protein were observed between normal and cancer tissues as well as amongdifferent normal or carcinoma tissues. Conclusion: ST13 protein is a cytoplasmic molecule with anapparent Mr of 50 000. The protein is expressed in colorectal and other epithelial tissues. Theexpression level of the protein is down-regulated in colorectal cancer and varies among differentnormal and/or carcinoma tissues. Comparison of cDNA sequences and protein characteristics indicatesthat ST13 protein and hsp70-interacting protein (Hip) are same proteins, raising the possibilitythat ST13 protein is involved in the development of colorectal cancer through Hsp70 molecularchaperone machinery.

Hoxc13 Expression Pattern in Cashmere Goat Skin During Hair Follicle Development

Hoxc13 has an important role in controlling hair formation. In this study, we examine the Hoxc13 RNA expression pattern of skin during embryo development. The result indicated that changes of the Hoxe13 gene expression and thickness of skin have a similar trend during hair follicle morphogenesis. In interpreting these results, we investigated whether the regulation motifs is in Hoxc13 intron, which is a 5.4 kb fragment. To blast with other mammals, we found a very conservative region in all mammal animals and two regions in livestock, such as cow, sheep, horse, dog, and so on, which are not in other Hox genes. We have examined putative pre-miRNA in this region, providing an entry point for elucidating currently unknown mechanisms that are required for regulating quantitative levels of Hoxc13 gene expression.

Expression of ST13 in colorectal cancer and adjacent normal tissues

AIM: To investigate the in situ expression of suppressionof tumorigenecity 13 (ST13) mRNA in both colorectal cancer and adjacent normal tissues.METHODS: Colorectal cancer cell lines SW1116, SW620and CoLo205 were enrolled to confirm the feasibility of the in situ hybridization procedure. Seven colorectal cancer and adjacent normal tissues were included for RNA-RNA in situ hybridization.RESULTS: The expression of ST13 in the seven normal colon tissues was positive and the positive signals appeared in mucosal cells. Only three of the seven colorectal cancer tissues had positive hybridization signals that appeared in adenocarcinoma cells.CONCLUSION: The expression of ST13 decreases in colorectal cancer tissue compared with that in adjacent normal tissue. ST13 is mostly expressed in colorectal epithelia and adenocarcinoma cells.

Expression and significance of MMP-7,c-Jun and c-Fos in rats skin photoaging

Objective:To investigate the expression and significance of the MMP-7,c-Jun and c-Fos in rat photoaging skin.Methods:A total of 45 SD rats were randomly divided into control group,model group,natural recovery group,physiological saline injection group and dermal pluripotent stem cells transplantation(DMSCs group),model group,natural recovery group,physiological saline injection group.DMSCs were treated with UV lamp irradiation to establish light aging skin model.Rats were then sacrificed after model prepared,no treatment was processed in the natural recovery group.Saline injections was adopted in saline group,DESCs group was treated with DESCs transplantation.Rats were sacrificed after 4 weeks.The expression of MMP-7,c-Jun and c-Kos were detected using the immunohistocheniical metluxl.Results:In model group,MMP 7positive expression was higher than that in the other 4 groups,but without statistically difference(P>0.05);c-Jun,c-Fos expression were higher than that in the control group and DESCs group(P<0.05),there was no significant difference comparing natural recovery group with physiological saline injection group(P>0.05).Conclusions:MMP-7,c-Jun and c-Fos can be used as diagnosis indicators in the early stage of light aging,and they jointly participate in its development.DMSCs transplants is effective in treating light aging skin.

Effect of Salvia Miltiorrhiza on Expression of the MMP-2 9 in Tissue of Early Stage Acute Lung Injury in Rats with Severe Acute Pancreatitis

Objective:To investigate the effect of salvia miltiorrhiza on expression of the MMP-2、9 and TIMP-1、TIMP-2 in tissue of acute lung injury of severe acute pancreatitis(SAP).Methods:MMP-2、9 expression and changes of the lung were measured after the SAP rats were induced by retrograde injection of 5%sodium tauocholate into hepatopancreatic duct.The changes of those parameters were also measured after salvia miltiorrhiza was injected intramuscularly just after induction of SAP.Results:The level of MMP-2、9 in pancreas and lung in SAP group were significantly higher than those in sham;The level of MMP-2、9 in salvia miltiorrhiza group were significantly lower than those in SAP group. Conclusion:MMP-2、9 were overexpressed in Acute lung injury (ALI) induced by SAP, salvia miltiorrhiza downregulates MMP-2、9 expression and decreased injury of lung tissue.

Vibramycin inhibits the expression of MMP-2 and the invasiveness of PC-3 cells in vitro

Objective To study the inhibition of vibramycin on the expression of matrix metalloproteinase-2 (MMP-2) and the invasiveness of androgen-independent prostatic carcinoma cell line PC - 3 in vitro. Methods Immunohistochemistry stain and transwell chamber were used to investigate the expression of MMP-2 in different concentration of vibramycin treated PC-3 cells and the invasive ability of different concentration of vibramycin treated PC-3 cell. Results The positive rate of MMP-2 inJune 2003 Vol12 No2 PC-3 cells was decreased at a concentration of 5 mg/L of vibramycin and decreased dramatically at the concentration of 10 mg/L. The cells moved throuth the membrane was(82. 0 ± 4.6)/field in the control group, while decreased to(26.1 ±3.6),(7.2 ±2.2) and(3.3± 0.7)/field in 5,10 and 20 mg/L vibramycin treated PC-3 cell respectively. Conclusion Vibramycin can inhibit the invasiveness and metastatasis of PC-3 cells, the mechanism of which is related to the inhibition of MMP-2 in PC-3 cell.10refs.

Quantitative expression of MMP-2 and FN in high metastatic and low metastatic cell lines of breast cancer

To analyze the relation of matrix metalloproteinase-2(MMP-2) and Fibronection (FN) mRNA expression with metastasis of breast cancer and elucidate the role of MMP-2 and FN in breast cancer metastasis.Methods The expression of MMP-2 and FN mRNA in breast cancer cell lines was detected by fluorescence-quantitative RT-PCR.The expression of MMP-2 and FN protein was detected by Western blots.Results The expression of MMP-2 and FN mRNA was down-regulated in high metastatic cell lines MDA-MB-231,MDA-MB-435,but up-regulated in low metastatic cell lines MDA-453,T47D,SK-BR-3 and non-metastatic cell line MCF-7,ZR-75-30.The protein expression of MMP-2 and FN was up-regulated in high mestastic cell lines,and down-regulated in low metastatic cell lines.Conclusion The mRNA and protein expression of MMP-2 and FN was related with breast cancer metastasis.The mRNA expression of MMP-2 and FN is feed-back regulated with protein expression.6 refs,4 figs,2 tabs.

侵袭性牙周炎患者牙龈组织中 MMP-1、MMP-8和 MMP-13蛋白的表达

目的:检测侵袭性牙周炎(AgP)患者牙龈组织中基质金属蛋白酶(MMP)-1、MMP-8及MMP-13的表达,探讨三者在AgP发病中的可能作用和意义。方法:采用免疫组化法检测30例AgP患者和20例健康对照牙龈组织中MMP-1、MMP-8及MMP-13蛋白的表达。结果:健康对照牙龈组织中MMP-1、MMP-8及MMP-13均有弱表达;AgP患者牙龈组织中MMP-1和MMP-8蛋白的表达高于健康对照(Z=3.980和2.990,P<0.05)。结论:MMP-1、MMP-8蛋白可能在AgP的发生中起重要作用。

MMP-1、MMP-13在食管鳞癌中的表达及与浸润转移的关系

目的研究食管鳞癌组织中基质金属蛋白酶-1(MMP-1)、MMP-13蛋白的表达情况及其与浸润转移之间的关系。方法应用免疫组织化学Elivision法检测90例食管鳞癌组织、16例重度不典型增生组织及28例正常食管黏膜组织中MMP-1、MMP-13蛋白的蛋白表达情况。结果在食管鳞癌、不典型增生及正常食管黏膜组织中MMP-1蛋白的阳性表达率分别为81.11%(73/90)、81.25%(3/16)和17.86%(5/28),MMP-13蛋白的阳性表达率分别为82.2%(74/90)、25%(4/16)、21.43%(6/28),三者差异有统计学意义(P<0.05)。MMP-1、MMP-13蛋白表达与临床分期、浸润深度及淋巴结转移有关(P<0.05)。MMP-1和MMP-13在食管鳞癌中表达呈正相关(r=0.537,P<0.05)。结论 MMP-1和MMP-13在食管鳞癌的发生、发展、浸润、转移过程中可能发挥重要作用,可能作为食管癌判断预后的重要标记。

非小细胞肺癌中MMP-9和MMP-13的表达及意义

目的分析MMP-9和MMP-13在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的表达及相关性,探讨二者与NSCLC临床病理特征及预后的关系。方法采用免疫组化SP法检测88例NSCLC和18例癌旁正常肺组织中MMP-9、MMP-13的表达。结果 MMP-9与MMP-13在NSCLC中的阳性率均高于癌旁正常肺组织(P<0.05)。MMP-9表达与患者年龄、分化程度、淋巴结转移有关(P<0.05);MMP-13表达与分化程度、淋巴结转移有关(P<0.05)。NSCLC中MMP-9与MMP-13的表达呈正相关(P<0.05)。MMP-9和MMP-13阳性组的生存率均低于阴性组,差异有显著性(P<0.05);且肿瘤大小、淋巴结转移和MMP-13阳性表达与NSCLC的预后密切相关(P<0.05)。结论 MMP-9与MMP-13均与NSCLC的侵袭、转移及预后密切相关,MMP-13可能主要通过激活MMP-9途径来参与NSCLC的侵袭、转移。

肺腺癌中MMP-9、MMP-13、HIF-1α表达与临床病理特征、EGFR突变及预后的关系

目的探讨MMP-9、MMP-13、HIF-1α在肺腺癌中的表达,分析其与肺腺癌临床病理特征、EGFR突变及预后的关系。方法采用免疫组化SP法检测629例肺腺癌组织中MMP-9、MMP-13、HIF-1α的表达,并选取癌旁正常组织、肺炎性假瘤样增生各50例作为对照。采用实时荧光PCR法检测629例肺腺癌EGFR的突变,并对患者进行随访。结果肺腺癌组中MMP-9、MMP-13、HIF-1α的表达均高于对照组(P<0.001)。MMP-9表达与患者淋巴结转移及肿瘤大小有关(P<0.05);MMP-13表达与患者吸烟史、TNM分期、淋巴结转移及肿瘤大小有关(P<0.05);HIF-1α表达与患者吸烟史、淋巴结转移有关(P<0.05);但三者表达均与EGFR突变无关(P>0.05)。经Kaplan-Meier生存分析结果显示:MMP-9、MMP-13和HIF-1α阳性组12、24、36个月的累积生存率均明显低于阴性组;COX多因素生存分析结果显示:EGFR突变、MMP-9、MMP-13、HIF-1α的表达、肿瘤大小、TNM分期均是肺腺癌患者生存状况的独立危险因素(P<0.05)。结论肺腺癌组织中MMP-9、MMP-13、HIF-1α存在过表达,其表达均与患者淋巴结转移有关,但与EGFR突变无关。MMP-9、MMP-13、HIF-1α呈阳性、EGFR无突变患者生存率低,且均为肺腺癌患者生存状况的独立危险因素。

腺相关病毒介导的klotho基因表达对去势大鼠骨Runx2及MMP-13表达的影响

目的探讨腺相关病毒介导的klotho(KL)基因表达对去势骨质疏松大鼠的调控作用。方法 SD雌性大鼠随机分为假手术组(S组)和手术组,外科去势术后12周再随机分为模型组(O组)、17β-雌二醇组(E组)、KL基因组(KO组)和空质粒组(GO组),实验12周后处死。取股骨、胫骨测骨密度;冰冻切片及免疫组化法观察肾KL荧光及KL蛋白表达;RT-PCR和免疫组化法检测骨Runx2、MMP-13 mRNA及蛋白表达;HE染色观察骨组织形态学变化。结果 KO组和E组骨密度高于O组和GO组(P<0.05);KO组大鼠肾有小鼠KL基因特异性表达;与O组相比,KO组Runx2 mRNA表达明显上调,MMP-13 mRNA表达显著下调(P<0.05);免疫组化分析KO组Runx2吸光度值为411±96,显著高于O组的353±50(P<0.05);KO组MMP-13吸光度值为397±84,显著低于O组的656±89(P<0.05)。KO组、E组和S组大鼠骨小梁排列紧密,连接成网,形态结构较完整,明显优于O组和GO组。结论 KL基因表达上调可减缓去势大鼠骨质疏松症的发展及骨组织微结构的破坏,提示KL基因可能在骨质疏松症的发展中扮演重要角色。

温针灸对不同时间点KOA模型兔关节软骨中MMP-1、MMP-13表达的影响

目的:观察温针灸(warming moxibustion)对兔膝骨关节炎(knee osteoarthritis,KOA)软骨细胞中基质金属蛋白-1、13(matrix metalloproteinase-1、13,MMP-1、MMP-13)表达的影响,明确温针灸治疗KOA的作用机制。方法:选取70只新西兰大耳兔,其中10只作为空白对照组,剩余随机分为模型2周组、模型4周组、模型6周组、温针灸2周组、温针灸4周组、温针灸6周组。采用右后肢伸直位石膏固定制备KOA兔模型,模型2、4、6周组不予处理,温针灸2、4、6周组分别给予温针灸治疗2周,治疗结束后取材。采用免疫组化及RT-PCR检测各组治疗前后关节软骨中MMP-1、MMP-13蛋白及mRNA的表达。结果:与空白对照组比较,模型2周组、模型4周组、模型6周组软骨中MMP-1、MMP-13表达明显升高(P<0.05),且各组间比较,差异有统计学意义(P<0.05)。与相应模型组比较,温针灸治疗后,MMP-1、MMP-13的表达降低(P<0.05),且各组间比较差异有统计学意义(P<0.05)。结论:温针灸可降低KOA兔软骨中MMP-1、MMP-13的表达,说明温针灸治疗KOA可能与其抑制MMP-1、MMP-13的表达有关。

光动力疗法对兔耳增生性瘢痕组织中MMP-9,MMP-13和TIMP-1的影响

目的研究ALA-PDT治疗对兔耳增生性瘢痕中基质金属蛋白酶(MMP)-9,MMP-13和金属蛋白酶组织抑制物(TIMP)-1的变化,探讨ALA-PDT治疗对兔耳增生性瘢痕的初步治疗机制。方法将实验动物分为正常组、阴性对照组、高浓度ALA-PDT治疗组、低浓度ALA-PDT治疗组和PDT治疗组,对治疗组分别进行干预治疗,1次/周,共3次,于治疗后1月、2月和3月采集样本,进行Western blot实验。结果治疗组中MMP-9,MMP-13在ALA-PDT治疗后1月、2月和3月的增生性瘢痕表达水平明显高于阴性对照组,差异有统计学意义(P<0.01);而治疗组中TIMP-1表达水平明显低于阴性对照组,差异有统计学意义(P<0.01);高浓度治疗组与低浓度治疗组之间比较差异有统计学意义(P<0.05)。结论 ALA-PDT治疗对兔耳增生性瘢痕的治疗是有效的,其治疗机制可能是通过逆转MMP-9,MMP-13和TIMP-1之间的失衡状态实现的。

IL-1β和MMP-13在兔骨关节炎模型软骨和滑液中的表达

目的评价兔膝关节内侧半月板损伤制作骨关节炎(OA)模型的方法的可行性,探讨白细胞介素-1β(IL-1β)和金属蛋白酶-13(MMP-13)在兔骨关节炎模型软骨和滑液病变中的作用机制。方法新西兰大白兔40只,随机分为实验组(n=30)和对照组(n=10),于术后2、6、12周观察股骨髁关节软骨的病理变化并进行评分;用免疫组化方法检测关节软骨中IL-1β和MMP-13的表达情况;用酶联免疫吸附法测定关节液中IL-1和MMP-13的含量。结果实验组和对照组在大体和病理观察不同时间点关节软骨评分及HE染色差异均有统计学意义(P<0.05)。免疫组织化学检测结果显示IL-1β在两组中均有表达,实验组和对照组2周时细胞分数差异无统计学意义,6、12周差异有统计学意义(P<0.05)。MMP-13在对照组关节软骨细胞中未见表达,实验组的关节软骨细胞中可见MMP-13蛋白的表达,差异有统计学意义(P<0.05)。关节滑液中IL-1β表达和在软骨中表达一致。结论采用损伤兔膝关节半月板的方法可建立合理的动物OA模型;IL-1β和MMP-13在OA发病过程中表达水平有明显变化,需要进一步的临床研究来探讨其是否可以作为早期诊断OA的指标之一。

臭氧联合玻璃酸钠对骨性关节炎患者关节液中MMP-13的影响

目的:探讨臭氧联合玻璃酸钠对骨性关节炎患者关节液中基质金属蛋白酶13(matrix metro politanate 13,MMP-13)的影响。方法:90例患者按就诊顺序随机平均分为A组为医用臭氧组,B组为玻璃酸钠组,C组为医用臭氧+玻璃酸钠组。在每次治疗前及治疗结束后1周及6个月行疼痛视觉模拟评分(Visual analogue scale,VAS)及临床疗效评分,检测关节液中MMP-13的含量。结果:A组患者治疗后VAS显著下降(P<0.05),MMP-13含量显升高(P<0.05);B组患者在治疗后VAS和临床疗效评分与治疗前无变化(P>0.05),MMP-13含量降低(P<0.05)。C组患者治疗后VAS降低(P<0.05),MMP-13含量降低(P<0.05)。结论:医用臭氧联合玻璃酸钠可以降低关节液中MMP-13的含量,具有一定程度的关节软骨保护作用,临床治疗效果良好,可以作为治疗膝骨性关节炎的一个方法在临床推广应用。

MMP-13、VEGF在三阴性乳腺癌中的表达及其临床意义

目的探讨基质金属蛋白酶-13(MMP-13)、血管内皮细胞生长因子(VEGF)在三阴性(ER-、PR-、HER2-)乳腺癌发生、发展过程中的临床意义。方法应用免疫组织化学SP法检测MMP-13、VEGF在60例三阴性乳腺癌、36例HR型乳腺癌(ER+/PR+、HER-2-)、28例HER2型乳腺癌(ER-、PR-、HER2+)和12例乳腺纤维腺瘤中的表达情况,并分析其与临床病理参数的关系。结果三阴性乳腺癌中MMP-13、VEGF的表达显著高于乳腺纤维腺瘤和HR型乳腺癌(P<0.005),而且MMP-13在三阴性乳腺癌中的表达显著高于HER2型乳腺癌(P<0.005);三阴性乳腺癌中MMP-13、VEGF阳性表达与腋窝淋巴结转移、临床分期有关(P<0.05),而与患者年龄、肿瘤大小无关(P>0.05)。相关分析显示,在三阴性乳腺癌、HR型乳腺癌及HER2型乳腺癌中MMP-13与VEGF的表达均呈正相关(r=0.522,r=0.482,r=0.571;P<0.01)。结论 MMP-13和VEGF与三阴性乳腺癌的发生、发展有着密切关系,可作为评估三阴性乳腺癌浸润和转移的重要生物学指标。