养心颗粒对近似不稳定型心绞痛家兔模型基质金属蛋白酶-9的影响

目的通过观察养心颗粒对不稳定型心绞痛家兔颈总动脉粥样硬化斑块基质金属蛋白酶-9(MMP-9)表达的影响,探讨其治疗不稳定型心绞痛的确切作用机制。方法以球囊一次性损伤家兔颈动脉结合高脂饮食喂养,8周后将28只家兔随机分为模型组、养心颗粒组、异山梨酯(消心痛)组和阿托伐他汀组,每组各7只,另取7只家兔为空白组,于10周末进行药物触发的方法复制成近似不稳定型心绞痛动物模型。用免疫组织化学法检测MMP-9的表达,采用HE染色观察模型血管病理形态学变化。结果模型组MMP-9表达明显高于空白组(P<0.0 1);与模型组相比,养心颗粒组和阿托伐他汀组均可下调MMP-9的表达(P<0.01),养心颗粒组的MMP-9表达显著低于消心痛组(P<0.05),而养心颗粒组和阿托伐他汀组MMP-9表达差异无统计学意义。病理研究结果表明,除空白组外,各组均出现血管内皮损伤,血管内膜增厚,脂肪沉积明显,脂质斑块形成。各治疗组颈总动脉内皮细胞轻度减少,内膜轻度增厚,内膜下脂质沉积减少。结论以益气养心安神法组成的中药复方制剂养心颗粒可以下调近似不稳定型心绞痛模型MMP-9的表达,这可能是其防治不稳定型心绞痛的作用机制之一。

多发性硬化复发、缓解期血清MMP-9、TIMP-1含量的变化

目的探讨血清基质金属蛋白酶-9(MMP-9)及其组织抑制剂-1(TIMP-1)水平与缓解-复发型多发性硬化(RRMS)的关系并评价其临床意义。方法选择RRMS复发期患者28例、缓解期患者20例,并选择正常体检者30例为对照组。用酶联免疫吸附法(ELISA)检测血清MMP-9、TIMP-1水平,并进行统计学分析。结果 RRMS复发期患者血清MMP-9水平、MMP-9/TIMP-1比值较对照组显著升高(P<0.01),较缓解期患者也有明显升高(P<0.05),TIMP-1水平较对照组降低明显(P<0.05),但与缓解期患者比较差别无统计学意义(P=0.655)。RRMS复发期患者血清MMP-9水平、MMP-9/TIMP-1比值与EDSS评分无相关性。结论血清MMP-9含量、MMP-9/TIMP-1比值与RRMS的炎症活动密切相关,可作为监测RRMS活动性的血清学指标。

Expressions of MMP-2,-9,TIMP-1,-2,-3 mRNA in Rat Uterus during Estrous Cycle

Zymography and in situ hybridization were used to investigate matrixmetalloproteinase -2, -9 (MMP -2, MMP-9) activities and expressions of MMP -2, -9 and TIMP1, -2, -3 (tissue inhibitors of matrix metallo-proteinases) mRNA in the rat uterus during estrouscycle. The relative activity was semiquanted by using densitometric analysis. The MMP-2(67 kDa) activity in every stage during estrpus cycle was detected by zymography. MMP-2activity was highest at proestrus; higher at estrus and metaestrus; lowest at diestrus. Throughin situ hybridization, MMP -2, -9, TIMP -1~ -3 mRNA mainly in hasal stroma cells of uterineendometrium were detected. The positive signals of MMP -2 and -9 mRNAs in hasal stromacells were shown stronger at proestrus, estrus and metaestrus while they showed the weakest atdiestrus. The expression of MMP -2 mRNA coincided with MMP -2 activity change. MMP-2and -9 mRNAs were also highly expressed in uterine circular muscle at estrus. Weak signals ofMMP -9 mRNA were detected in uterine luminal and glandular epithelial cells at estrus.TIMP -1 mRNA in hasal stroma cells was shown as the strongest expression at estrus andmetaestrus; stronger at proestrus and the weakest at diestrus. TIMP-2 mRNA in basal stromacells was stronger at estrus and diestrus; weaker at proestrus and metaestrus. TIMP -1 and -2mRNAs were also highly expressed in uterine luminal and glandular epithelial cells at estrus.TIMP -3 mRNA in hasal stroma cells revealed the strongest expression at estrus; stronger atdiestrus and metaestrus and showed the weakest at proestrus. The mRNA was also highlyexpressed in uterine circular muscle at estrus. In short, our present results provide evidencethat MMP -2, -9 and TIMP -1~ -3 were involved in rat uterine endometrium reconstructionduring estrous cycle.

Alterations of glutathione S-transferase and matrix metalloproteinase-9 expressions are early events in esophageal carcinogenesis

AIM: To investigate the role of glutathione S-transferase (GST) and matrix metalloproteinase-9 (MMP-9) expres-sions in the development and progression of reflux es-ophagitis-Barrett’s metaplasia-dysplasia-adenocarcinoma sequence in the esophagus.METHODS: GST and MMP-9 expressions were analyzed in 51 paraffin-embedded tissue samples by immunohisto-chemistry including patients with reflux esophagitis (n = 7), Barrett’s metaplasia (n = 14), Barrett and esophagi-tis (n = 8), Barrett and dysplasia (n = 7), esophageal adenocarcinoma (n = 8) and a control group without any histological changes (n = 7). Immunostaining was determined semiquantitatively. Statistical analysis with one-way ANOVA, LSD test and correlation analysis were performed. P value of < 0.05 was considered significant.RESULTS: GST expression was significantly higher while MMP-9 expression was significantly lower in control group compared to Barrett’s metaplasia and the other groups. No major changes were observed between Bar-rett, esophagitis, and Barrett and concomitant esophagi-tis. Barrett and concomitant dysplasia, and adenocarci-noma revealed a significant lower expression of GST and higher levels of MMP-9 compared to all other groups. Adenocarcinoma showed almost no expression of GST and significantly higher levels of MMP-9 than Barrett and concomitant dysplasia. Alterations of GST and MMP-9 were inversely correlated (r = - 0.82).CONCLUSION: Decreased GST and increased ex-pression of MMP-9 in Barrett’s metaplasia-dysplasia-adenocarcinoma sequence as compared to normal tissue suggest their association with esophageal tumorigenesis. Loss of GST and gain of MMP-9 in Barrett with dyspla-sia compared to non-dysplastic metaplasia indicate that these alterations may be early events in carcinogenesis. Quantification of these parameters in Barrett’s esopha-gus might be useful to identify patients at higher risk for progression to cancer.

Expressions of inducible nitric oxide synthase and matrix metalloproteinase-9 and their effects on angiogenesis and progression of hepatocellular carcinoma

AIM： To determine the expressions of inducible nitric oxide synthase （iNOS） and matrix metalloproteinase-9 （MMP-9） in hepatocellular carcinoma （HCC） and to investigate the relationship between iNOS and MMP-9 expression and their effects on angiogenesis and progression of HCC.METHODS： In this study, we examined iNOS, MMP-9, and CD34 expression in specimens surgically removed from 32 HCC patients and 7 normal liver tissues by immunohistochemical staining. Meanwhile, microvessel density （MVD） was determined as a marker of angiogenesis by counting CD34-positive cells. RESULTS： The positive rates of iNOS and MMP-9 expression were 71.88% （23/32） and 78.13% （25/32） in HCC. MMP-9 expression was significantly correlated with tumor size, capsule status, TNM stage, and risk of HCC recurrence （P = 0.032, P= 0.033, P= 0.007, and P= 0.001, respectively）. There was also a significant relationship between iNOS expression and capsule status and risk of HCC recurrence （P = 0.049 and P = 0.004, respectively）, but no correlation between iNOS expression and tumor size and TNM stage. There was a positive association between MVD and TNM stage and risk of HCC recurrence （P = 0.037 and P = 0.000, respectively）. The count of MVD was significantly different in different iNOS and MMP-9 immunoreactivity groups （F= 17.713 and 17.097, P= 0.000 and P = 0.000, respectively）. The examination of Spearman＇s rank correlation coefficient showed that there was a significant positive correlation between MVD and iNOS, MMP-9 immunoreactivity （r = 0.754 and 0.751, P= 0.000 and P=-0.000, respectively）. There was also a significant association between MMP-9 and iNOS expression in HCC （P = 0.010）. CONCLUSION： Nitric oxide （NO） produced by iNOS could modulate MMP-9 production and therefore contribute totumor cell angiogenesis and invasion and metastasis in HCC. The strong expression of iNOS and MMP-9 in HCC may be helpful in evaluating the recurrence of HCC, predicting poor prognosis. For patients with strong expression of MMP-9 and iNOS, the optimal treatment scheme needs to be selected.

Clinical significance of matrix metalloproteinase-9 expression in esophageal squamous cell carcinoma

AIM: To evaluate the expression of matrix metalloproteinase-9 (MMP-9) and its clinical significance in esophageal squamous cell carcinoma (ESCC). METHODS: The expression of MMP-9 in 208 cases of ESCC was detected by immunohistochemistry (IHC) and its clinical significance in ESCC especially the relationship with the clinicopathological parameters was analyzed. RESULTS: The percentage of positive cases for MMP-9 detected by IHC was 49.0%. MMP-9 was mainly expressed in the cytoplasm of cancer cells especially in the invasive front. Only weak expression was detected in the stromal cells and no expression in non-cancerous mucosa. The expression of MMP-9 was positively correlated with poorer differentiation (P= 0.001<0.01), existence of vessel permeation (P= 0.027<0.05) and lymph node metastasis (P=0.027<0.05). CONCLUSION: The expression of MMP-9 correlates with the cancer cell differentiation, vessel permeation and lymph node metastasis. It may be a novel biomarker for the diagnosis and treatment of ESCC.

Expression and correlation of CD44v6, vascular endothelial growth factor, matrix metalloproteinase-2, and matrix metalloproteinase-9 in Krukenberg tumor

AIM： To explore the expression and correlation of CD44v6, vascular endothelial growth factor （VEGF）, matrix metalloproteinase （MMP）-2 and matrix metalloproteinase （MMP）-9 in Krukenberg and primary epithelial ovarian carcinoma. METHODS： The expressions of CD44v6, VEGF, MMP-2 and MMP-9 were detected by immunohistochemical method in 20 cases of normal ovarian tissues, 38 cases of Krukenberg tumor and 45 cases of primary epithelial ovarian carcinoma. RESULTS： The expression of CD44v6 （primary epithelial ovarian carcinoma tissue vs normal ovarian tissue： χ^2= 4.516, P= 0.034; Krukenberg tumor tissue vsnormal ovarian tissue： χ^2 = 19.537, P = 0.001） and VEGF （primary epithelial ovarian carcinoma tissue vs normal ovarian tissue： P = 0.026; Krukenberg tumor tissue vs normal ovarian tissue： χ^2 = 22.895, P = 0.001） was significantly higher in primary epithelial ovarian carcinoma tissue and Krukenberg tumor tissue than in normal ovarian tissue. The positive expression rate of MMP-2 and MMP-9 was 0% in the normal ovarian tissue. The positive expression rate of CD44v6 （ χ^2= 10.398, P= 0.001）, VEGF （ χ^2= 13.149, P = 0.001）, MMP-2 （ χ^2 = 33.668, P = 0.001） and MMP-9 （ χ^2= 38.839, P = 0.001） was remarkably higher in Krukenberg tumor than in primary epithelial ovarian carcinoma. The correlation of CD44v6, VEGF, MMP-2, and MMP-9 was observed in primary epithelial ovarian carcinoma and Krukenberg tumor. CONCLUSION： CD44v6, VEGF, MMP-2, and MMP-9 are involved in ovarian carcinoma, gastric cancer and Krukenberg tumor. Detection of CD44v6, VEGF, MMP-2 and MMP-9 may contribute to the diagnosis of ovarian carcinoma, gastric cancer, and Krukenberg tumor.

The relationship between matrix metalloproteinases-9 and CD15s antigen in angiogenesis of gastric carcinoma

Objective： To investigate the relationship between matrix metalloproteinases-9 （MMP-9） and sialyl Lewis X （CD15s） antigen in invasion and metastasis of gastric carcinoma. Methods： Expression of CD105, MMP-9 and CD15s in 47 cases of gastric carcinoma undergone radical surgery were evaluated by SP immunohistochemical staining using the respective monoclonal antibody. The microvessel density （MVD） marked with CD105 was detected. Correlation between MVD and MMP-9, CD15s was also statistically analyzed. Results： The MVD in both MMP-9 and CD15s positive expression group was higher significantly than that in MMP-9 or CD15s positive expression alone group, and it was also significantly higher than that in both MMP-9 and CD15s negative expression group. Conclusion： MMP-9 is a marker of invasion and CD15s is a marker of metastasis in gastric carcinoma. Combining detection of MMP-9 and CD15s has certain clinical significance for diagnosis, treatment and assessing the prognosis for gastric caner.

Expressions of cyclooxygenase-2 and matrix metalloproteinase-9 in cervical carcinoma and their clinical significance

Objective: To investigate the expressions of cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 in cervical carcinoma and their clinical significance. Methods: Immunohistochemistry SP method was used to detect the expres- sions of COX-2 and MMP-9 in 72 cases of invasive carcinoma of cervix (ICC) and 16 cases of normal cervical epithelium remote from tumor (NCE). The relationships between the expressions of COX-2, MMP-9 in ICC and some characteristics relating to clinical pathology of cervical carcinoma such as histological grading, lymph node metastasis, stromal invasion and FIGO stage were analyzed statistically. Results: The rates of the positive expressions of COX-2 and MMP-9 in ICC were significantly higher than those in NCE. COX-2: 88.9% (64/72) in group ICC and 12.5% (2/16) in group NCE, P = 0.000; MMP-9: 94.4% (68/72) in group ICC and 43.8% (7/16) in group NCE, P = 0.000. The expression of COX-2 was positively correlated with lymph node metastasis (r = 0.296, P = 0.012) and stromal invasion (r = 0.257, P = 0.029). The expression of MMP-9 was positively correlated with FIGO stage (r = 0.329, P = 0.005) and histological grading (r = 0.351, P = 0.003). The expression of COX-2 was positively correlated with the expression of MMP-9 in ICC (r = 0.297, P = 0.011). Conclusion: The overexpressions of COX-2 and MMP-9 were closely related to the invasion and growth of cervical carcinoma. The tissue with the overexpression of COX-2 had strong invasion ability. COX-2 and MMP-9 had synergistic effect on proliferation, invasion and metastasis of cancer cells. Detecting the coexpression of COX-2 and MMP-9 may be of value in further understanding the biological behavior and predicting the prognosis of cervical carcinoma.

Contrary Regulation of TIMP-1 and MMP-9 by Hepatocyte Growth Factor Antibody after Lung Injury

Objective To study the influence of hepatocyte growth factor (HGF) antibody on the lung expression level of matrix metalloproteinases-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1). Methods Thirty male Wistar rats were randomly divided into 3 groups: control group, model group, and intervention group. Endotoxin was intratracheally infused in the model and intervention groups. HGF antibody was injected in the rats of the intervention group from day 1 to day 14, while the same volume of saline was injected in the control group. The rats were sacrificed on day 28 after endotoxin treatment. The amounts of MMP-9 mRNA and TIMP-1 mRNA were measured by reverse transcription-polymerase chain reaction, and protein expression levels of MMP-9 and TIMP-1 were measured by immunohistochemistry. Results In the model group, both mRNA and protein expression levels of TIMP-1 were significantly increased, the same as MMP-9. In the intervention group, the increase of TIMP-1 was remarkably reduced compared with the model group, while the mRNA and protein expression levels of MMP-9 were still increased. Conclusion HGF activity may accelerate the repair of lung injury through contrary regulating the expression levels of TIMP-1 and MMP-9.

Serum metrix metalloproteinase-9 combined with homocysteine, IL-6, TNF-α, CRP, HbA11c and lipid profile in the incipient diabetic nephropathy with or without macrovascular diseases

Objective ： To evaluate the changes of serum matrix metalloproteinase-9 （romp-9） in patients of incipient diabetic nephropathy with or without macrovascular disease and to analyze the factors associated with homocysteine（hcy）, interleukin-6（IL-6）, tumor necrosis factor-alpha （TNF-α）, highly sensitive C-reactive protein （hsCRP）, HbAlc and lipid profile in those patients in order to know whether this marker or other factors are more important to induce diabetic macrovascular disease. Methods： Type 2 diabetes mellitus（T2DM） subjects with incipient diabetic nephropathy with or without macrovascular disease were selected for participation and divided into 2 groups. The patients in group 1 （n= 38） used insulin, and patients in group 2 （n=34） were treated with an oral antidiabetic drug. Then serum mmp-9, hey, IL-6 and TNF-α in these patients were measured, and compared to the healthy subjects as control （n= 16）. The results were analyzed by SPSS13. Results： Serum romp-9 and hcy of the patients having incipient diabetic nephropathy with macrovascular disease were higher than that of patients without macrovascular disease （P〈0.01）. For insulin-injected patients, whether they accompanied with macrovascular diseases or not, the serum levels of romp-9, hcy, IL-6 and TNF-α were all lower, but no significant statistics compared with non-insulin used patients or the healthy subjects. The serum level of romp-9 was more correlated with the serum hcy in antidiabetic drug used patients. （P〈0. 000） Conclusion： The serum level of romp-9 plays an important role of pathogenesis in the macrovascular disease in the incipient diabetic patients, and the serum level of hcy also can reflect the severely degree of macrovascular disease in these patients, insulin can reduce these markers.

Elevated plasma level of matrix metalloproteinases-9 and its significance in patients with abdominal aortic aneurysms

Objective: To investigate the changes of the plasma level of MMP-9 (Matrix Metalloproteinase-9, MMP-9) in the patients with abdominal aortic aneurysms (AAAs) before and after the treatment, and evaluate the significance of MMP-9 in the pathogenesis of AAAs. Methods: Blood samples of 35 patients with AAAs and 10 patients with the arterial occlusive diseases (AODs) , which enrolled into the Vascular Surgery Center of Colonge University Hospital from February to August of 2002, were collected before and one month after surgical repair or less-invasive endovascular exclusion. The plasma concentrations of MMP-9 of all the collected samples were measured by means of enzyme-linked immunosorbent assay(ELISA), and compared between the two groups patients at different time point. Results: The mean plasma concentration of MMP-9 of AAAs was significantly higher than that of AODs prior to treatment [(90.3±9.1) ng/ml vs (23.6±7.3) ng/ml, P<0.05], and no apparent difference was showed in the patients with AODs [(23.6±7.3) ng/ml vs (25.3±5.8) ng/ml, P>0.05)] before and after the surgical bypass operation. However, in the patients with AAAs the plasma concentration of MMP-9 was apparently decreased one month after the surgical repair or endovascular exclusion compared with before [(28.6±8.4) ng/ml vs (90.3±9.1) ng/ml, P<0.05)]. No meaningful difference of the mean plasma MMP-9 concentration was seen between two groups after the both being successfully treated [(28.6±8.4) ng/ml vs (25.3±5.8) ng/ml, P>0.05]. Conclusion: Apparent elevation of plasma concentration of MMP-9 in the AAAs and its dramatic decrease after being treated implicated that MMP-9 might play an important role in the formation and development of AAAs. Meanwhile, to investigate the changes of MMP-9 level of AAAs could provide an practical way to facilitate the earlier diagnosis and long term surveillance for AAAs. More importantly, pharmacologic prevention and treatment of AAAs, in which the MMP-9 serves as effective target, might be possible in the future.

RNF13： a novel RING-type ubiquitin ligase over-expressed in pancreatic cancer

Protein ubiquitination by E3 ubiquitin ligases plays an important role in cancer development. In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. Immunohistochemical analysis of pancreatic ductal adenocarcinoma （PDAC） and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples （41.7%, 30/72）, and its expression was significantly associated with histological grading （P= 0.024）. In addition, RNF13 was detected in precancerous lesions： tubular complexes in chronic pancreatitis （CP） and pancreatic intraepithelial neoplasia （PanIN） （79.3%, 23/29 and 62.8%, 22/35, respectively）. Moreover, RNF13 staining was significantly correlated with Tenascin-C expression （P = 0.004） in PDAC samples, further supporting the role of RNF13 in cancer progression. Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells increased invasive potential and gelatinolytic activity by matrix metalloproteinase-9. Taken together, these findings reveal that RNF13 is a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis; ubiqui-tin-mediated modification of proteins by RNF13 may participate in pancreatic cancer development.

EFFECTS OF GENISTEIN ON INVASION AND MATRIX METALLOPROTEINASE ACTIVITIES OF HT1080 HUMAN FIBROSARCOMA CELLS

Effects of genistein on invasion and matrix metalloproteinase activities were investigated in HT1080 human sarcoma cells.Invasion of HT1080 cells through reconstituted basement membrane was inhibited when the cells were treated with 100 μ mol/L and 200 μ mol/L genistein.At the same concentrations,genistein not only suppressed latent forms of matrix metalloprotinese 2 and 9(MMP 2 and MMP 9) to convert into active forms,but also increase dramatically the tissue inhibitor of metalloproteinase(TIMP 1) mRNA contents and reverse the imbalance of MMPs and TIMPs.However,expressions of MMP 2 and MMP 9 were not significantly affected.Suppression of MMP activation and increase of TIMP 1 expression will decrease matrix degradation by MMPs,and consequently inhibit invasions of the cells.These results emphasized the existence of the imbalance between MMPs and TIMPs in tumor invasion and metastasis formation.The value of genistein as a drug for antiinvasion and anti metastasis chemotherapy was suggested.

Effects of carbon dioxide pneumoperitoneum on metastasis of ovarian carcinoma cell line SKOV_3

The long suspicion of the potential harm of carbon dioxide （CO2） pneumoperitoneum exists in laparoscopic cancer surgery. For better understanding of this problem, we targeted this study at the effects of CO2 pneumoperitoneum on the invasive ability of ovarian carcinoma cell line and the possible mechanism within it. To study the effects of CO2 pneumoperitoneum on carcinoma cell, SKOV3 cells were divided into 2 groups, respectively exposed to pneumoperitoneal CO2-insuffiation and normal conditions. To study the possible mechanism, SKOV3 cells were divided into 3 groups, one was exposed to CO2 pneumoperitoneum, one to N2 and the other to normal conditions served as control. The in vitro adhesive and invasive ability of the cells was analyzed through 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium bromide （MTT） assay and Boyden filters metastasis model; the expressions of vascular endothelial growth factor C （VEGF-C） and matrix metalloproteinase 9 （MMP9） were determined by reverse transcription polymerase chain reaction（RT-PCR）, and Western blot. We found that the adhesive ratio of SKOV3 cells exposed to CO2 was significantly higher than that of the control group; cells exposed to CO2 invaded the matrigel with a greater number （P〈0.01）; the expression of VEGF-C exposed to both CO2 and N2 was significantly increased compared with control group （P〈0.05）; the MMP9 expression level of CO2 group was higher than that of N2 group, P〈0.05. We concluded that carbon dioxide pneumoperitoneum may improve the adhesive and invasive ability of ovarian carcinoma cell line in vitro and CO2 can also be an independent factor to stimulate the expression of MMP9.

IL-5、IL-12、MMP-9和TIMP-1在不同海拔地区慢性阻塞性肺疾病合并慢性肺源性心脏病患者血清中的表达及意义

目的 通过检测中、高海拔地区慢性阻塞性肺疾病（COPD）合 并 慢 性 肺 源 性 心 脏 病（CPHD）患者血清白介素5（IL-5）、IL-12、基质金属蛋白酶9（MMP-9）和金属蛋白酶组织抑制剂1（TIMP-1）的变化水平并探讨其临床意义。方法 选取 2012年 9月至 2014年 8月青海省西宁市 （中度海拔,2260m）COPD合并 CPHD患者和健康体检者各30例、青海省玉树州 （高海拔,3680m）相同疾病患者 22例和健康体检者24例,采用 酶联免疫吸附试验法分别测定其血清IL-5、IL-12、MMP-9及 TIMP-1的表达水平并进行比较。结果 中、高海拔病例组血清IL-5水平均低于其对照组（t=4.069、4.530,P 值均〈0.01）,高海拔病例组和对照组的血清IL-5水平均高于中海拔病例组和对照组 （t=14.118、20.284,P 值均〈0.01）;中、高海拔病例组血清IL-12水平均高于其对照组 （t=5.967、2.274,P 值均〈0.05）,高海拔病例组血清IL-12水平低于中海拔病例组 （t=2.598,P 〈0.05）;中、高海拔病例组血清 MMP-9水 平 均 高 于 其 对 照 组 （t=21.032、10.308,P 值 均 〈0.01）,高海拔病例组和对照组的血清 MMP-9水平高于中海拔的病例组和对照组 （t=6.991、10.771,P 值均〈0.01）;中海拔病例组血清 TIMP-1水平高于其对照组 （t=6.626,P 〈0.01）,中海拔病例组和对照组血清 TIMP-1水平均高于高海拔病例组和对照组 （t=7.015、3.292,P 值均 〈0.01）,高海拔病例组和对照组的血清 TIMP-1水平差异无统计学意义 （t=0.228,P〉0.05）。结论 中、高海拔地区 COPD合并 CPHD患者血清中IL-5、IL-12、MMP-9及 TIMP-1表达水平发生了相应的变化,其水平变化可能参与了气道免疫炎症反应和气道重塑;同时,上述炎症因子在不同海拔地区之间表达具有一定差异,考虑高原缺氧环境更易诱发和加重机体炎症反应。

Glucosamine reduces blood-brain barrier disruption by inhibiting the expression of matrix metalloproteinase-9 in experimental autoimmune encephalomyelitis rats

We investigated the effects of glucosamine(GS) on blood-brain barrier(BBB) function and matrix metalloproteinase-9 (MMP-9) expression in rats with experimental autoimmune encephalomyelitis(EAE).Animals were randomly divided into three groups,among which the EAE and GS groups were immunized with complete antigen and pertussis toxin,and the adjuvant group was immunized with complete Freund's adjuvant and pertussis toxin.Rats were treated by peritoneal injection of GS 180 mg/(kg·d) in the GS group and peritoneal injection of phosphate-buffered saline 4.5 mL/(kg·d) in the EAE and adjuvant groups.We proposed to assess the integrity of BBB by calculating cerebrospinal fluid to serum albumin quotient(QA) on days 6,8,10,12,14,16 and 18 post-immunization.At the same time,the brains and spinal cords were removed for MMP-9 immunohistochemical staining. Experiments demonstrated that in the EAE group,QA value and MMP-9 expression were highly elevated and up-regulated and correlated to disease severity.Moreover,there was statistically significantly positive correlation between QA value and MMP-9 expression.In the GS group,we observed that the mean disease onset date was delayed,the incidence and mean score of symptom were suppressed at the peak phase of disease(P<0.05).Furthermore,QA value and MMP-9 expression in the GS group showed stronger inhibition when compared with those of the EAE group(P<0.05).Our study showed that GS would reduce the BBB breakdown and leukocyte trafficking by inhibiting the production of MMP-9 and mitigate EAE.

短暂性脑缺血发作病人的脑缺血耐受作用及血清MMP-9水平的变化

目的评价短暂脑缺血发作(TIA)是否具有脑保护作用并探讨其机制。方法选择2000～2004年142例脑梗死病人。脑梗死前有 TIA 病史者分为3组,A 组30例,TIA 持续时间10 min;B 组30例,TIA 持续时间10～20min;C 组30例,TIA 持续时间20min;52例无 TIA 病史者列为 D 组。采用欧洲脑卒中神经功能缺损评分标准(ESS),对各组患者人院后24 h、21d 及发病后3个月进行神经功能评分;应用 Pullicino 等公式对各组病人发病21d 时进行脑梗死体积计算;应用酶联免疫吸附法测定各组病人发病12、24、48、72、96 h 时血清 MMP-9的水平。结果 A、B、C 3组病人神经功能评分明显高于 D 组,B 组病人明显高于 A、c两组;发病21 d 时,A、B、C 3组病人梗死体积明显小于 D 组,B 组最明显;每组患者血清 MMP-9的水平呈动态变化即发病12 h 开始增高,48 h 达到高峰,96 h 时恢复到正常水平。B 组病人在不同时间点血清 MMP-9的含量明显低于 A、C 两组,且明显低于 D组。结论缺血耐受在脑梗死前合并 TIA 的患者可能发挥作用。其作用可以与 MMP-9的表达减少有关。

Contribution of epigenetics in diabetic retinopathy

Diabetes has become the epidemic of the 21 st century, and with over 90% patients with diabetes becoming at a risk of developing retinopathy, diabetic retinopathy has emerged as a major public health concern. In spite of cutting edge research in the field, how retina and its vasculature are damaged by the diabetic milieu remains ambiguous. The environmental factors, life style or disease process can also bring in modifications in the DNA, and these epigenetic modifications either silence or activate a gene without altering the DNA sequence. Diabetic environment up- or downregulates a number of genes in the retina, and emerging research has shown that it also facilitates epigenetic modifications. In the pathogenesis of diabetic retinopathy, the genes associated with important enzymes(e.g., mitochondrial superoxide dismutase, matrix metalloproteinase-9 and thioredoxin interacting protein) and transcriptional factors are epigenetically modified, the enzymes responsible for these epigenetic modifications are either activated or inhibited, and the levels of micro RNAs are altered. With epigenetic modifications taking an important place in diabetic retinopathy, it is now becoming critical to evaluate these modifications, and understand their impact on this slow progressing blinding disease.

Effect of warming Yang and removing blood stasis method on matrix metalloproteinases/tissue inhibitor metalloproteinases levels secreted by cultured endometrial cells from patients with endometriosis

OBJECTIVE:To investigate the effect of Chinese medicines using the warming Yang and removing blood stasis method on levels of matrix metalloproteinases(MMPs)/tissue inhibitor metalloproteinases(TIMPs) secreted by cultured endometrial cells from patients with endometriosis.METHODS:Ectopic and eutopic endometrial cells obtaind from 15 endometriosis patients were cultured in vitro,and divided randomly into five groups:high dose;moderate dose;low dose;nemestran;blank control.The three dose groups were treated with a decoction prepared according to the principle of warming Yang and removing blood stasis;nemestran and 0.9%NaCI were administered to the nemestran group and balnk control group,respectively.Eutopic endometrial cells obtaind from 10 hysteromyoma patients were cultured in vitro,as the normal control group,0.9%NaCI were administered to the normal control group.Cell culture supernatants were collected and levels of matrix metalloproteinase-1(MMP-1),matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),tissue inhibitor metalloproteinase-1(TIMP-1) and tissue inhibitor metalloproteinase-2(TIMP-2) detected by enzyme-linked immuno sorbent assay(ELISA).RESULTS:Compared with the normal control group,levels of MMP-1,MMP-2,and MMP-9 in eutopic and ectopic endometrium cell supernatants in the blank control group were increased,whereas levels of TIMP-1 and TIMP-2 were decreased(P <0.05).Compared with the blank control group,levels of MMP-1 and MMP-2 in ectopic and eutopic endometrium cell supernatants cultured in low-dose,middle-dose,and high-dose groups were decreased,whereas levels of TIMP-1 and TIMP-2 were increased significantly(P < 0.05).CONCLUSION:The warming Yang and removing blood stasis method affects expression of MMPs andTIMPs.