Name: a mice model of chlorhexidine gluconate-induced peritoneal damage
Uploaded: 2022-04-28T14:45:00
Description: Watch this Scientific Journal Video about A Mice Model of Chlorhexidine Gluconate-Induced Peritoneal Damage at JoVE.com

我们衷心感谢Shin-Han Tseng对这项研究的批判性讨论和部分执行。本研究得到了台湾E-DA医院研究基金和国立成功大学的EDAHP110003和NCKUEDA110002的支持。

所有小鼠实验均由E-DA医院/义寿大学实验动物中心批准，并根据“实验动物护理和使用指南”（NRC，美国2011）进行处理。雄性C57BL / 6小鼠，7-8周龄，用于本研究。
1. 化学制剂
<ol><li>通过在15%乙醇中稀释0.1%葡萄糖酸氯己定（CG，见 材料表）来制备化学刺激物。</li>
</ol>2. 动物治疗
<ol><li>指定三只小鼠作为对照组。每隔?...

<ol>
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C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vitamin+D+can+ameliorate+chlorhexidine+gluconate-induced+peritoneal+fibrosis+and+functional+deterioration+through+the+inhibition+of+epithelial-to-mesenchymal+transition+of+mesothelial+cells.">Vitamin D can ameliorate chlorhexidine gluconate-induced peritoneal fibrosis and functional deterioration through the inhibition of epithelial-to-mesenchymal transition of mesothelial cells.</a> BioMed Research International. 2015, 595030 (2015).</li><li>Nakamoto, H., Kawaguchi, Y., Suzuki, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Is+technique+survival+on+peritoneal+dialysis+better+in+Japan.">Is technique survival on peritoneal dialysis better in Japan.</a> Peritoneal Dialysis International. 26 (2), 136-143 (2006).</li><li>Schaefer, F., Klaus, G., Muller-Wiefel, D. 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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modification+of+the+masson+trichrome+technique+for+routine+laboratory+purposes.">Modification of the masson trichrome technique for routine laboratory purposes.</a> The American Journal of Pathology. 14 (2), 237-243 (1938).</li><li>Cheng, F. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+application+of+magnetite+nanoparticle-mediated+vitamin+D3+delivery+for+peritoneal+dialysis-related+peritoneal+damage.">Novel application of magnetite nanoparticle-mediated vitamin D3 delivery for peritoneal dialysis-related peritoneal damage.</a> International Journal of Nanomedicine. 16, 2137-2146 (2021).</li><li>Ross, A., Willson, V. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Basic and Advanced Statistical Tests: Writing Results Sections and Creating Tables and Figures. , 13-16 (2017).</li><li>Suga, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preventive+effect+of+pirfenidone+against+experimental+sclerosing+peritonitis+in+rats.">Preventive effect of pirfenidone against experimental sclerosing peritonitis in rats.</a> Experimental and Toxicologic Pathology. 47 (4), 287-291 (1995).</li><li>Ishii, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+experimental+sclerosing+encapsulating+peritonitis+model+in+mice.">An experimental sclerosing encapsulating peritonitis model in mice.</a> Nephrology Dialysis Transplantation. 16 (6), 1262-1266 (2001).</li><li>Nishino, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antisense+oligonucleotides+against+collagen-binding+stress+protein+HSP47+suppress+peritoneal+fibrosis+in+rats.">Antisense oligonucleotides against collagen-binding stress protein HSP47 suppress peritoneal fibrosis in rats.</a> Kidney International. 64 (3), 887-896 (2003).</li><li>Mishima, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhanced+expression+of+heat+shock+protein+47+in+rat+model+of+peritoneal+fibrosis.">Enhanced expression of heat shock protein 47 in rat model of peritoneal fibrosis.</a> Peritoneal Dialysis International. 23 (1), 14-22 (2003).</li><li>Kushiyama, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+liposome-encapsulated+clodronate+on+chlorhexidine+gluconate-induced+peritoneal+fibrosis+in+rats.">Effects of liposome-encapsulated clodronate on chlorhexidine gluconate-induced peritoneal fibrosis in rats.</a> Nephrology Dialysis Transplantation. 26 (10), 3143-3154 (2011).</li><li>Nishino, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Involvement+of+lymphocyte+infiltration+in+the+progression+of+mouse+peritoneal+fibrosis+model.">Involvement of lymphocyte infiltration in the progression of mouse peritoneal fibrosis model.</a> Renal Failure. 34 (6), 760-766 (2012).</li><li>Lua, I., Li, Y., Pappoe, L. 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本研究通过腹腔注射CG提出了小鼠PD模型，结果显示该模型存在腹膜纤维化和功能恶化，模拟PD患者的病情。
协议中有几个关键步骤。首先，为了进行CG或NS的腹腔注射，必须使用镊子拾取小鼠的腹壁皮肤，以防止穿刺引起的腹膜内器官损伤。其次，在收集腹壁腹膜进行组织学分析时，必须避免腹腔注射损伤的区域。
在腹膜纤维化的几种实验动物模型中，?...

腹膜透析（PD）是一种肾脏替代疗法。但是，PD存在无法解决的问题。例如，长期PD治疗可引起腹膜损伤，最终导致超滤失败和治疗停止1，2，3，4，5，6。腹膜纤维化是最严重的并发症之一7，8。腹膜纤维化的特征是间质内细胞外基质的沉积和积累，以及腹膜9，10的新血管生成和血管病变。
这些腹膜变化的主要原因是复发性腹膜炎和透析液的非生物相容性，它们是高渗、高葡萄糖、低pH和葡萄糖降解产物积累11，12。因此，合适的动物实验模型可以帮助研究人员更好地研究PD治疗过程中腹膜的生理和病理变化。因此，建立动物PD模型对于提高PD技术和透析疗效，延长患者生存期具有重要意义。本研究旨在通过腹膜内（ip）注射葡萄糖酸氯己定（CG）来生成PD小鼠模型，如前所述13，14。与其他PD动物模型相比，该PD小鼠模型简单，易于使用且可行。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.9% Normal Saline</td>
			<td>Y F CHEMICAL CORP., New Taipei City, Taiwan</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>10% neutral buffered formalin</td>
			<td>Taiwan Burnett International Co., Ltd., Taipei City, Taiwan</td>
			<td>00002A</td>
			<td></td>
		</tr>
		<tr>
			<td>Automatic biochemical analyzer</td>
			<td>Hitachi Ltd., Tokyo, Japan</td>
			<td>Labospect Series 008</td>
			<td>for determining glucose concentration</td>
		</tr>
		<tr>
			<td>Chlorhexidine digluconate solution, 20% in H2O</td>
			<td>Sigma-Aldrich, MO, USA</td>
			<td>C9394</td>
			<td>diluted to 0.1% with 15% ethanol for injection</td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Avantor Performance Materials, LLC, PA, USA</td>
			<td>BAKR8006-05</td>
			<td>diluted to 15% with normal saline for working concentration</td>
		</tr>
		<tr>
			<td>Glucose (Dianeal)</td>
			<td>Baxter International, Inc., IL, USA</td>
			<td>FNB9896</td>
			<td>Commercial dialysis solution (4.25%)</td>
		</tr>
		<tr>
			<td>GraphPad Prism 8.0</td>
			<td>GraphPad Software, Inc., CA, US</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>L-type Glu 2 assay</td>
			<td>FUJIFILM Wako, Japan</td>
			<td>461-32403</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine 20</td>
			<td>Juily Pharmaceutical Co., Ltd., New Taipei City, Taiwan</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>Zoletil 50</td>
			<td>Virbac Laboratories, Carros, France</td>
			<td>-</td>
			<td></td>
		</tr>
	</tbody>
</table>

a mice model of chlorhexidine gluconate-induced peritoneal damage

腹膜纤维化是腹膜透析（PD）的重要并发症。为了调查和解决这个问题，需要适当的PD动物模型。本协议建立了葡萄糖酸氯己定（CG）诱导的腹膜纤维化模型，该模型模拟PD患者的状况。 腹膜内注射0.1%的CG在15%乙醇中3周（每隔一天给药），在雄性C57BL / 6小鼠中总共9次诱导腹膜纤维化。然后在第22天进行腹膜功能测试。处死小鼠后，收获腹壁壁腹膜和肝脏内脏腹膜。在Masson的三色染色后进行显微镜分析时，它们更厚，纤维化更多。超滤速率降低，葡萄糖质量转运表明CG诱导的腹膜通透性增加。这样建立的PD模型在改善PD技术、透析疗效和延长患者生存期方面具有应用价值。

在图1A，B中，在Masson三色染色17下，腹壁壁壁腹膜明显增厚且纤维化更多，表明在CG暴露组中，腹膜纤维化比对照盐水组（NS）更严重。在图2A，B中，肝脏表面的内脏腹膜也明显增厚且纤维化，从而证明在CG暴露组中，腹膜纤维化比对照盐水组（NS）更严重。在图3A中，CG组的超滤速率降低?...

Watch this Scientific Journal Video about A Mice Model of Chlorhexidine Gluconate-Induced Peritoneal Damage at JoVE.com

A Mice Model of Chlorhexidine Gluconate-Induced Peritoneal Damage

本协议建立了葡萄糖酸氯己定（CG）诱导的腹膜纤维化的腹膜透析（PD）小鼠模型。与其他PD动物模型相比，目前的模型简单易用。

葡萄糖酸氯己定诱导腹膜损伤的小鼠模型

Peritoneal fibrosis is an important complication of peritoneal dialysis (PD). To investigate and address this problem, an appropriate animal model of PD ...

建立腹膜透析（PD）动物模型非常重要。我们希望它能提高PD治疗的技术和疗效，最终提高患者的生存率。与其他腹膜透析动物模型相比，该腹膜透析小鼠模型简单，易于使用且可见。合适的腹膜透析动物模型可以帮助研究人员研究PD治疗过程中的腹膜、生理和病理变化。该程序的演示将由我实验室的研究助理Fang-Ling Liao进行演示。首先，通过将0.1%葡萄糖酸氯己定稀释在15%乙醇中来制备化学刺激物。接下来，指定三只小鼠作为对照组，每隔一天腹膜内注射每公斤0.9%生理盐水一毫升，持续三周，共九次。然后将三只小鼠分配到腹膜纤维化组，并通过腹膜内注射先前制备的0.1%葡萄糖酸氯己定溶液以每公斤体重12.5微升的剂量诱导腹膜纤维化。对于腹膜功能测试，准备含有4.25%葡萄糖的透析溶液。然后使用注射器抽取0.5毫升透析液样品。准备唑莱地尔和甲苯噻嗪的注射剂，并通过肌内注射麻醉小鼠。以每20克体重两毫升的剂量进行透析溶液的腹膜内安装。在腹部中线做一个小切口，用注射器收集主要的腹膜内液。30分钟后，在剑突下方的腹部中线进行垂直切口，并打开小鼠的腹部，用注射器收集残留的腹膜内液。接下来，测量干净干燥的棉签的重量。将棉花放入小鼠腹腔吸收残留的腹腔内液，然后将其取出。再次测量棉花重量。Masson的三色染色显示，葡萄糖酸氯己定暴露组腹壁的顶腹膜明显比对照组更厚且纤维化。葡萄糖酸氯己定暴露组肝表面内脏腹膜也明显较对照盐水组较对照组更厚且纤维化，表明葡萄糖酸氯己定暴露组腹膜纤维化更严重。葡萄糖酸氯己定暴露组表现出超滤率降低和葡萄糖质量转运增加，表明葡萄糖酸氯己定暴露小鼠腹膜通透性增加在进行腹膜内注射程序时，使用镊子并抠开小鼠的腹部皮肤，以防止穿刺引起的腹膜内器官损伤。

Research

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术中的微妙的子宫内膜异位症的检测：A新型范式为相关联的与的的腹膜的完整性之亏损盆腔疼痛的的的的检测和治疗的

Endometriosis is a  common disease affecting 40 to 70% of reproductive-aged women with chronic  pelvic pain (CPP) and/or infertility. The purpose of this ...

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一个简单的临界尺寸股骨缺损模型小鼠

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腹膜内卵巢癌转移的定量

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Myocardial infarction induced by coronary artery ligation has been used in many animal models as a tool to study the mechanisms of cardiac repair and ...

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Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

小鼠加速动脉硬化的诱导："电线损伤"模型

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

A Tissue Displacement-based Contusive Spinal Cord Injury Model in Mice

基于组织位移的小鼠脊髓损伤模型

Producing a consistent and reproducible contusive spinal cord injury (SCI) is critical to minimizing behavioral and histological variabilities between ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

A Model of Glaucoma Induced by Circumlimbal Suture in Rats and Mice

Circumlimbal 缝合法诱导大鼠和小鼠青光眼模型的建立

The circumlimbal suture is a technique for inducing experimental glaucoma in rodents by chronically elevating intraocular pressure (IOP), a well-known ...