变态反应性疾病儿童血浆肥大细胞羧肽酶和类糜蛋白酶含量的检测

目的：测定变态反应性疾病患儿血浆中肥大细胞羧肽酶和类糜蛋白酶的含量,评价其在变态反应性疾病诊断中的意义。方法：采用ELISA法检测59例变态反应性疾病儿童和53例健康儿童血浆中肥大细胞羧肽酶和类糜蛋白酶水平。结果：变态反应性疾病患儿血浆羧肽酶和类糜蛋白酶含量分别为1.089±0.752 ng/mL、0.905(0.375~2.318)ng/mL,显著高于健康儿童［0.593±0.380 ng/mL、0.454(0.097~1.077) ng/mL］,差异有统计学意义（P<0.05)。变态反应性疾病患儿血浆羧肽酶水平与类糜蛋白酶水平呈显著正相关(r=0.684,P<0.01)。结论：变态反应性疾病儿童血浆中肥大细胞羧肽酶和类糜蛋白酶水平增高,提示二者可作为变态反应性疾病的诊断的有意义的指标。     

Dermal fibrosis in male pattern hair loss: a suggestive implication of mast cells

A relationship has been suggested between mast cells (MCs) and male pattern hair loss (MPHL), because of histological evidence of perifollicular fibrosis and increased mast cell numbers. Two paired punch biopsies were taken from balding vertexes and non-balding occipital promontory areas of ten patients with MPHL (Ludwig–Hamilton IIIv to IV) and from five normal subjects aged from 20 to 35 years. Masson trichrome and Victoria blue staining were performed to observe collagen frameworks and elastic fiber structures. Numbers of immunoreactive MCs stained with anti-tryptase or anti-chymase antibody were counted. It was found that collagen bundles were significantly increased in balding vertexes than in non-balding occiput scalp skin. A near 4-fold increase in elastic fibers was observed in both vertex and occiput scalp skins with MPHL versus controls. Total numbers of MCs (tryptase-positive) in site-matched scalp samples were about 2-fold higher in MPHL subjects than in normal controls. Percentage elastic fiber (%) was found to be relatively well-correlated with tryptase and chymase-positive MCs. These findings suggest that accumulated MCs might be responsible for increased elastic fiber synthesis in MPHL, and indicate that future investigations are warranted.     

Velcade影响K562细胞基因表达谱的分子机制

目的 应用生物信息学方法探索Velcade影响K562细胞基因表达谱的分子机制.方法 提取并扩增Velcade处理组和溶剂对照组的K562细胞RNA,与22KAgilent Human 1A基因芯片杂交,用Agilent Feature Extraction软件采集扫描数据,再以GeneSifter,GATHER等基因芯片数据分析工具,对差异表达基因进行基因本体分类、KEGG通路分析、蛋白互作网络分析和文献挖掘.结果 获得228个差异表达基因,其中上调84个,下调144个.下调糜酶1基因幅度最大.对数比值达10.80倍,干扰素α-21基因也下调2.31倍.本体分类显示,衰老过程、白细胞活动等过程显著增强:KEGG通路分析显示,JAK-STAT信号通路,自然杀伤细胞介导的细胞毒作用和抗原处理与提呈等通路受显著影响;蛋白互作网络揭示泛素依赖的蛋白质降解通路、抗原提呈和免疫反应、JAK-STAT信号通路等处于网络中重要位置;文献挖掘显示差异表达基因与白血病、细胞凋亡、细胞周期、蛋白酶体、抑制剂、衰老和IκB等关键词高度关联.结论 Velcade可能通过抑制NF-κB和JAK-STAT等促进细胞存活的信号通路,增强细胞毒作用,诱导肿瘤细胞凋亡;Velcade还可能参与抗原加工与提呈、免疫反应、炎症反应等过程;糜酶1基因可能是Velcade发挥抗肿瘤作用的关键靶标.     

Usefulness of serum mast cell–specific chymase levels for postmortem diagnosis of anaphylaxis

Chymase, a serine protease, is stored mainly in secretory granules of human mast cells. Serum chymase concentration was examined in 8 autopsy cases with anaphylaxis as well as in 104 control cases without anaphylaxis. It was detected in all 8 cases with anaphylaxis (range 3–380 ng/ml, mean 89.8 ng/ml), while it was detected in only 2 of the 104 controls and was below a detectable level (<3 ng/ml) in the other 102. Serum tryptase levels are known to be a diagnostic indicator of anaphylaxis, therefore the relationship between serum chymase and tryptase levels was investigated in the 8 cases of anaphylactic death; a significant positive correlation was found (r=0.826, p=0.011). Furthermore, chymase was shown to be quite stable in serum. These results showed that measurement of serum chymase levels might be an additional tool for postmortem diagnosis of anaphylaxis.     

肺动脉高压大鼠T型和TC型肥大细胞的改变

目的:建立SD大鼠高肺血流并用野百合碱注射所致肺动脉高压模型,观察肺组织中肥大细胞及肥大细胞两种亚型的变化,探讨肥大细胞在肺动脉高压中的机制。方法:大鼠腹主动脉-下腔静脉造瘘,术后7天一次性腹腔注射野百合碱60mg/kg,制造肺动脉高压模型,35天后测定肺动脉压,取肺组织进行免疫组织化学染色,观察肥大细胞的变化,HE染色观察肺组织一般病理变化。结果:HE染色显示肺组织中动脉管壁明显增粗,有新生血管形成。平均肺动脉压力模型组较对照组升高(P<0.05);右心室/(左心室+室间隔)模型组较对照组增加(P<0.05),肥大细胞总数及类糜蛋白酶阳性的肥大细胞模型组较对照组增多(P<0.05)。结论:腹主动脉-下腔静脉造瘘合并野百合碱腹腔注射成功的建立了肺动脉高压的动物模型,肥大细胞在肺组织的聚集以及活化、肥大细胞亚型的改变可能是肺动脉高压形成的机制之一。     

肝组织中糜酶浓度在慢性肝炎肝纤维化中的意义

目的:为探讨肝组织中糜酶(Chymase)浓度在慢性病毒性肝炎肝纤维化中的意义。方法:采用酶联免疫吸附实验(ELISA)检测45例慢性肝炎肝组织中Chymase浓度,观察不同的纤维化分期(S1-S4),其肝组织中Chymase浓度;用免疫组织化学染色法观察Chymase单克隆抗体标记的肥大细胞分布情况。结果:慢性肝炎纤维化分期重的S3和S4患者,其肝组织中Chymase浓度(S3+S4=31.3±24.6ng/mg)明显高于纤维化轻的S1和S2患者(S1+S2=5.7±4.8ng/mg),P<0.01;免疫组化染色结果,Chymase标记的肥大细胞主要分布于纤维化旺盛的汇管区与类洞壁,其分布与纤维化部位相一致,且肝组织中Chymase浓度高的患者,其肝组织内Chymase标记的肥大细胞分布增多。结论:肝组织中Chymase浓度可能与慢性肝炎肝纤维化关系密切。     

Possible roles of mast cell-derived chymase for skin rejuvenation

The relationships between mast cell-derived chymase, angiotensin II, and extracellular matrix production in the skin after intense pulsed light (IPL) were clarified in hamsters. Dorsal areas of the hamsters were irradiated once or twice a week by IPL. The index of extracellular matrix production in the skin was defined as the depth stained with Azan-Mallory stain from the epidermis to the dermis at the point of maximum thickness. The index had significantly increased 7 days after IPL irradiation in sections treated once or twice with IPL compared with that of untreated control sections. The numbers of mast cells, chymase-positive cells, and angiotensin II-positive cells had also significantly increased in IPL-irradiated areas. Significant increases in chymase and angiotensin II activities were observed in the extracts obtained from IPL-irradiated skin. Mast cell-derived chymase may be involved via angiotensin II formation in the dermal extracellular matrix production that occurs after IPL irradiation.     

心脏糜酶对自发性高血压大鼠心肌纤维化的影响

目的观察心脏糜酶对自发性高血压大鼠(SHR)心肌组织胶原合成和心肌纤维化的影响.方法应用病理检查、计算机分析结合逆转录-聚合酶链式反应等方法,检测SHR应用糜酶抑制剂(Ch-Ⅰ)组、SHR组及WistarKyoto大鼠(WKY)组的收缩压、心肌胶原容积分数(CVF)、心肌血管周围胶原面积比(PVCA)和心肌糜酶及Ⅰ、Ⅲ型胶原mRNA表达.结果Chy-Ⅰ组心脏CVF、PVCA分别为(26.8±8.7)%和0.4±0.1,SHR组分别为(46.4±7.8)%和1.9±0.9,WKY组为(24.4±10.7)%和0.4±0.1,Chy-Ⅰ组比SHR组明显下降(P＜0.01),与WKY组无明显差别(P＞0.05).Chy-Ⅰ组心肌组织Ⅰ、Ⅲ型胶原和糜酶mRNA表达相对含量均明显低于SHR组(P＜0.01),与WKY组无明显差别(P＞0.05).应用Chy-Ⅰ后大鼠血压无改变.结论心肌组织糜酶参与胶原的合成,参与细胞外基质的形成和降解,促进自发性高血压大鼠心肌纤维化.     

Skin mast cell promotion of matrix remodeling in burn wound healing in mice: relevance of chymase

Abstract Inflammation, granulation, and collagen accumulation, which are observed in the wound healing process, occasionally lead to hypertrophic scarring. Several in vitro reports have suggested that skin mast cells (MCs) and their major protease, chymase, participate in the healing process as well as in fibrotic skin diseases. The present study examined the potential involvement of MCs and MC chymase in the healing of burns in mouse dorsal skin. The size of the burn wounds, density of the capillaries, collagen accumulation, MC number, and chymase activity were measured before and 1, 3, 7, and 14 days after burning. The healing process corresponded strongly with MC density and chymase activity in both acute and subacute phases. The maximum decrease in MC number and chymase activity occurred on day 3 when tissue loss due to necrosis was maximal. From day 7 to 14, the burn wounds retracted rapidly accompanied by increases in capillaries and collagen fibers, in correspondence with fast increments in MC numbers and chymase activity at the wound edges. The present results combined with previous in vitro results strongly support the contention that skin MC chymase plays a role in the normal wound healing process, and presumably in dermal fibrotic disorders. Received: 6 January 1998     

糜酶途径对犬血管平滑肌细胞产生血管紧张素Ⅱ的影响

目的:探讨糜酶途径对犬血管平滑肌细胞(VSMC)产生血管紧张素Ⅱ(AngⅡ)的影响。方法:通过组织块贴壁法培养犬动脉VSMC,分为5组:对照组、AngⅠ组、卡托普利组、糜酶抑素组、联合用药组。对照组中仅为空白培养液,其他各组均加入AngⅠ,卡托普利组、糜酶抑素组和卡托普利+糜酶抑素组再分别加入相应的干预药物。应用放射免疫法检测培养液中AngⅡ含量等。结果:本实验成功培养出犬VSMC,含AngⅠ各组的AngⅡ浓度均高于对照组(P0.05);卡托普利组的AngⅡ浓度减少,但与AngⅠ组比较差异无统计学意义(P0.05);糜酶抑素组明显低于AngⅠ组和卡托普利组(P0.05);卡托普利+糜酶抑素组作用更强一些,但与糜酶抑素组比较差异无统计学意义(P0.05)。结论:糜酶在犬VSMC的AngⅡ生成过程中起着重要作用,其抑制剂可抑制AngⅡ生成。