Effectiveness of conjunctival bleb scarring by knockdown of heat shock protein 47 in rat model

AIM:To evaluate the effectiveness of knock-down of heat shock protein 47(HSP47)on conjunctival bleb scarring in a rat model and its possible mechanism.METHODS:Male Sprague–Dawley rats were used for glaucoma filtration surgery(GFS)and were treated with either phosphate buffered solution,shControl,mitomycin C,or sh-HSP47 using a microsyringe immediately after GFS.The morphology of filtering blebs was observed postoperatively.The levels of HSP47 were analyzed at 2,5,8,and 11d after GFS via real‑time quantitative polymerase chain reaction(PCR)and Western blot.The silencing effect of HSP47,the expression of collagen I and III,and the potential signaling pathways of HSP47 during scarification were explored 11d post GFS.The protein levels of transforming growth factor-β1(TGF-β1),phospho-Smad2(pSmad2),phospho-Smad3(p-Smad3),and phospho-p38(p-p38)were also analyzed using Western blot.RESULTS:Sh-HSP47 treatment significantly prolonged the functional filtration bleb retention.The levels of HSP47 were increased significantly at 5,8,and 11d postoperatively compared to the control group(P<0.05,P<0.01,and P<0.001).The levels of HSP47 protein at day 11 postoperatively were significantly down-regulated after HSP47 silencing using sh-HSP47 adenovirus transfection(P<0.01).Expression levels of collagen I and III within the blebs were significantly reduced in the absence of HSP47(P<0.01).Moreover,the protein levels of TGF-β1,p-Smad2/3,and p-p38 were dramatically inhibited after treatment with sh-HSP47(P<0.01).CONCLUSION:The inhibitory effects of HSP47 knockdown on scarring after GFS have the potential to be an efficacious therapeutic option for the treatment of conjunctival bleb scarring.

不同品种仔猪运输应激后组织损伤与Hsp47表达

选取12头皮特兰和24头二花脸仔猪,体重达到(20±1)kg时,随机选取6头皮特兰和12头二花脸进行2h运输,其余仔猪作为对照组。采用HE染色、免疫组织化学以及酶联免疫吸附试验技术,通过对不同品种仔猪运输前后肝脏、肾脏、胃的损伤以及Hsp47的定位、定量观察,研究运输应激与Hsp47表达之间的关系。结果表明:2h运输应激后,二花脸和皮特兰仔猪各组织的实质细胞均有不同程度的损伤;Hsp47主要分布在肝脏的中央静脉和窦状隙内皮细胞、肾小球和远曲小管上皮细胞、胃的黏膜层和黏膜下层结缔组织内;在肾脏和胃组织中,皮特兰仔猪在运输后Hsp47含量增加(P<0.05),但二花脸仔猪的相应值未见显著差异,2个品种肝脏中Hsp47含量在运输前后的变化无差异。Hsp47在受检组织中尽管有表达增加的趋势,但2个不同品种仔猪组织中Hsp47含量升高的程度不同,说明二者对运输应激的敏感性不同,二花脸仔猪具有更强的抗应激能力。

血清碱性成纤维细胞生长因子（bFGF）及热休克蛋白47（HSP47）水平与房颤发病的相关性研究

目的：通过测定房颤患者血清bFGF及HSP47水平与房颤的相关性研究，探讨血清bFGF及HSP47水平在早期房颤患者中的作用及意义。方法：用酶联免疫法检测40例房颤患者的血清bFGF及HSP47的含量，其中持续性房颤及阵发性房颤各20例，另外选取20例窦性心律的健康者作为对照组，结果采用SPSS17．0软件进行统计学处理。结果：持续房颤组及阵发房颤组的血清bFGF及HSP47水平明显高于健康对照组（69．78±23．52μg／m3V828．05±2．41ng／mlvsl5．87±4．25ng／ml，150．3±29．10ng／mlVS109．23±8．18ng／mlvs64．42±18．65ng／ml，P〈0．01），且持续性房颤患者体内的血清bFGF及HSP47水平明显高于阵发性房颤组（69．78±23．52vs28．05±2．41，150．34±29．10vsl09．23±8．18，P〈0．01）。结论：房颤患者血清bFGF及HSP47水平上调且明显高于健康对照组，差异具有统计学意义（P〈0．01），bFGF及HSP47在房颤的发生和维持过程中可能发挥重要作用。

Cardiac fibroblast heat shock protein 47 aggravates cardiac fibrosis post myocardial ischemia-reperfusion injury by encouraging ubiquitin specific peptidase 10 dependent Smad4 deubiquitination

Despite complications were significantly reduced due to the popularity of percutaneous coronary intervention(PCI) in clinical trials, reperfusion injury and chronic cardiac remodeling significantly contribute to poor prognosis and rehabilitation in AMI patients. We revealed the effects of HSP47 on myocardial ischemia-reperfusion injury(IRI) and shed light on the underlying molecular mechanism.We generated adult mice with lentivirus-mediated or miRNA(mi1/133TS)-aided cardiac fibroblastselective HSP47 overexpression. Myocardial IRI was induced by 45-min occlusion of the left anterior descending(LAD) artery followed by 24 h reperfusion in mice, while ischemia-mediated cardiac remodeling was induced by four weeks of reperfusion. Also, the role of HSP47 in fibrogenesis was evaluated in cardiac fibroblasts following hypoxia-reoxygenation(HR). Extensive HSP47 was observed in murine infarcted hearts, human ischemic hearts, and cardiac fibroblasts and accelerated oxidative stress and apoptosis after myocardial IRI. Cardiac fibroblast-selective HSP47 overexpression exacerbated cardiac dysfunction caused by chronic myocardial IRI and presented deteriorative fibrosis and cell proliferation.HSP47 upregulation in cardiac fibroblasts promoted TGFβ1-Smad4 pathway activation and Smad4 deubiquitination by recruiting ubiquitin-specific peptidase 10(USP10) in fibroblasts. However, cardiac fibroblast specific USP10 deficiency abolished HSP47-mediated fibrogenesis in hearts. Moreover, blockage of HSP47 with Col003 disturbed fibrogenesis in fibroblasts following HR. Altogether, cardiac fibroblast HSP47 aggravates fibrosis post-myocardial IRI by enhancing USP10-dependent Smad4 deubiquitination,which provided a potential strategy for myocardial IRI and cardiac remodeling.

HSP47 Deletion Inhibits TGF-β1 Stimulation on Proliferation and Collagen Synthesis of Human Tenon Fibroblasts

Heat shock protein 47 (HSP47) is a collagen-specific molecular chaperone that is required for mo-lecular maturation of various types of collagens. Many studies have shown a close association be-tween increased expression of HSP47 and excessive accumulation of collagens in scar tissues of various human fibrotic diseases. However, the role of HSP47 in formation of scar after glaucoma filtration surgery is still unclear. In this study, we deleted the expression of HSP47 in human tent on fibroblasts (HTFs) by virus infection, and then the proliferation and collagen synthesis were compared between HSP47 deletion cells and control upon TGF-β1 stimulation. Our data showed that HSP47 deletion could significantly inhibit the proliferation and collagen synthesis of HTFs upon TGF-β1 stimulation, HSP47 gene suppression might be a novel method to against the formation of scar after glaucoma surgery.

热休克蛋白47在新月体肾炎中的作用研究

目的探讨热休克蛋白47(HSP47)在新月体肾炎细胞性新月体向纤维性新月体演变过程中的作用。方法选择1998年1月至2005年8月东南大学附属中大医院肾活检病例中8例新月体肾炎患者,通过免疫组化及免疫双染方法,观察 HSP47在细胞性新月体向纤维性新月体演变过程中的表达及其意义探讨。结果新月体肾炎细胞性新月体内 HSP47表达明显增加,而纤维性新月体内的表达减少;免疫双染显示新月体内大多数α-SMA 阳性细胞同时亦有 HSP47表达。结论新月体肾炎细胞性新月体 HSP47过度表达,或许通过帮助前胶原分子在细胞内处理及加工,参与细胞性新月体向纤维性新月体转分化过程,导致肾脏发生不可逆性损伤。