실험은 250-300g 무게 암컷 흰쥐에서 수행 하였다. 실험실 동물의 관리 및 사용 (실험 동물 자원 연구소, 과학의 국립 아카데미, 베데스다, MD, USA)와 대한 설명서에 명시된 기준을 준수 모든 동물의 절차는 의학 기관 동물 관리의 메이요 클리닉 대학에 의해 승인되었다 및 사용위원회 (IACUC). 1. 준비 <ol><li> 수술 전에 수술 도구를 압력솥. 다른 쥐의 여러 수술이 같은 날에 계획하는 경우, 각 동물 절차 후 악기를 세척하고 뜨거운 비드 살균기를 사용하여 소독. </li><li> 1 L / 분 O 2에서 4 % 이소 플루 란과 쥐를 마취. </li><li> 37 ° C의 체온을 유지하기 위해 조절 된 가열 패드로 쥐를 옮긴다. 1 L / 분 O 2에 1 ~ 2 %의 이소 플루 란으로 마취를 유지한다. </li><li> 진통 약물 (부 프레 놀핀 릴리스 0.6 ㎎ / ㎏을 지속) 스와 관리bcutaneously. </li><li> 절차를 수행하는 동안 건조를 방지하기 위해 눈 연고를 적용합니다. </li><li> 개복에 의한 체액 손실을 보상하기 위해, 수술 전에 0.9 % 생리 식염수를 피하에 10 ㎖ / kg을 투여하는 것이 중요하다. </li><li> 복부 영역을 면도하고 포비돈 - 요오드 및 70 % 에탄올 패드​​로 피부를 청소하십시오. </li></ol> 2. 수술 <ol><li> 진정의 깊이는 발가락 핀치에서 철수, 눈꺼풀 반사, 턱 톤, 호흡 속도 / 패턴으로 물리적 반사 신경을 모니터링하여 적절한 지 확인합니다. </li><li> 수술 용 메스 블레이드 10 호를 사용하여 작은 중간 선 절개를 통해 개복술 (길이 2-2.5 cm)를 수행합니다. </li><li> 면봉을 사용하여 복부의 오른쪽에 소장과 대장을 당겨 기관 촉촉한 유지하기 위해 0.9 % 생리 식염수에 적신 멸균 거즈로 커버. </li><li> 조심스럽게 AOR을 노출 위쪽으로 비장, 간, 위장과 췌장을 철회타 왼쪽 신장 동맥. </li><li> 수술 현미경의 도움으로 조심스럽게 위 왼쪽 신장과 혈관, 지방에서 왼쪽 신장 동맥과 무딘 해부 곡선 집게와 멸균 면봉와 주변의 결합 조직 아래 복부 대동맥을 분리합니다. <ol><li> 지방을 제거하는 측면 회전 운동과 결합 조직과 면봉을 제거하기 위해 선박의 길이를 따라 반복 개폐 동작 (무딘 절개)와 집게를 사용합니다. 참고 : 요정 대동맥 영역의 절개가 손상 될 수 있습니다 신경과 림프관으로 매우 민감한 단계입니다. 해부 과정에서 식염수로 촉촉한 동맥을 유지해야합니다. </li></ol></li><li> 대동맥 아래 4-0 실크 봉합사를 놓습니다. </li><li> 미세 혈관 클립을 사용하여, (그냥 장간막 동맥 이하) 및 신장 동맥 분기 아래에 위의 대동맥 클램프. </li><li> 왼쪽 kidn 수준에서 대동맥 천공24 G 정맥 카테터와 EY 동맥 분기 및 신장 동맥에 카테터를 진행합니다. 참고 :이 발생할 수 있습니다 신장 동맥을 통해 천자 등이 중요한 단계이다. </li><li> 카테터에 약액 또는 (500 μL까지) 식염수로 채워진 주사기를 연결하고 신장 관류. </li><li> 즉시 관류 후, 미세 혈관 클립 왼쪽 신장 정맥 왼쪽 요관을 고정하고 카테터를 제거합니다. 이어서, 대동맥 천공 영역에, 조직 접착제의 작은 방울, 흡수성 지혈제 젤라틴 스폰지 조각을 넣고 부드럽게 면봉으로 압력을 적용한다. </li><li> 동시에, 좌측 신장 동맥의 분기점 아래 복부 대동맥으로부터 클램프 해제. 5 분 후, 신장 혈관 및 요관에서 클램프를 놓습니다. </li><li> 조심스럽게 왼쪽 신장 동맥 분기 이상, 대동맥에서 클램프를 분리 한 신장 재관류를 허용합니다. 총 신 허혈은 7 분보다 더 오래 지속 없습니다. </li><l난&gt; 활성 출혈이 발생하지 않도록 확인하고 밀접하게 10 이상 분의 영역을 관찰합니다. </li><li> 감염을 예방하기 위해 4-0 흡수성 봉합사 및 연속적인 패턴을 사용하여, 두 개의 층 (근육과 피부)의 복부 절개를 닫는다. 연속 패턴 봉합 기술에 더하여, 또 다른 옵션은 열개 않도록 특히 체벽 폐쇄위한 간단한 인터럽트 기술을 사용하는 것이다. </li><li> 감염을 방지하기 위해 절개 영역에 국소 항생제 연고를 적용합니다. </li><li> 35-37 ° C에서 설정된 온도 범위 전체 복구 될 때까지 따뜻한 패드에 침구없는 관찰 케이지에 쥐를 전송합니다. 느슨한 침구 커버 (예 : 드레이프 또는 종이 수건)이나 동물이 완전히 질식 또는 침구의 구토물이기도를 막는 것을 방지하기 위해 복구 될 때까지 케이지에서 제거해야합니다. </li><li> 수술 후 몇 시간 동안 매 시간 후, 자발적으로 호흡 할 때까지 지속적으로 동물을 관찰한다. 진통 부 프레 놀핀 SR 72 시간을 다시 투여 량나중에 불편의 흔적은 같은 웅크 리고 앉게하고 초라한 혼수, 얼굴을 찡 그리기, 정상적인 활동을 재개하지으로 관찰하는 경우. </li><li> 모든 연구 종료 후, CO 2의 과다 복용의 흡입과 동물을 안락사와 생체의 신장 조직은 조직 학적 및 웨스턴 블로 팅 (5)로 분석 수확. </li></ol>

<ol>
	<li>Dejani, H., Eisen, T. D., Finkelstein, F. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revascularization+of+renal+artery+stenosis+in+patients+with+renal+insufficiency.">Revascularization of renal artery stenosis in patients with renal insufficiency.</a> Am. J. Kidney Dis. 36 (4), 752-758 (2000).</li><li>Kang, D. 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=Impaired+angiogenesis+in+the+remnant+kidney+model:+I.+Potential+role+of+vascular+endothelial+growth+factor+and+thrombospondin-1.">Impaired angiogenesis in the remnant kidney model: I. Potential role of vascular endothelial growth factor and thrombospondin-1.</a> J. Am. Soc. Nephrol. 12 (7), 1434-1447 (2001).</li><li>Kang, D. H., Hughes, J., Mazzali, M., Schreiner, G. F., Johnson, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+angiogenesis+in+the+remnant+kidney+model:+II.+Vascular+endothelial+growth+factor+administration+reduces+renal+fibrosis+and+stabilizes+renal+function.">Impaired angiogenesis in the remnant kidney model: II. Vascular endothelial growth factor administration reduces renal fibrosis and stabilizes renal function.</a> J. Am. Soc. Nephrol. 12 (7), 1448-1457 (2001).</li><li>Kang, D. 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=Role+of+the+microvascular+endothelium+in+progressive+renal+disease.">Role of the microvascular endothelium in progressive renal disease.</a> J. Am. Soc. Nephrol. 13 (3), 806-816 (2002).</li><li>Chade, A. R., Kelsen, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reversal+of+renal+dysfunction+by+targeted+administration+of+VEGF+into+the+stenotic+kidney:+a+novel+potential+therapeutic+approach.">Reversal of renal dysfunction by targeted administration of VEGF into the stenotic kidney: a novel potential therapeutic approach.</a> Am. J. Physiol.- Renal Physiol. 302 (10), F1342-F1350 (2012).</li><li>Eirin, A., 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=Changes+in+Glomerular+Filtration+Rate+After+Renal+Revascularization+Correlate+With+Microvascular+Hemodynamics+and+Inflammation+in+Swine+Renal+Artery+Stenosis.">Changes in Glomerular Filtration Rate After Renal Revascularization Correlate With Microvascular Hemodynamics and Inflammation in Swine Renal Artery Stenosis.</a> Circ.-Cardiovasc. Interv. 5 (5), 720-728 (2012).</li><li>Chade, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinct+Renal+Injury+in+Early+Atherosclerosis+and+Renovascular+Disease.">Distinct Renal Injury in Early Atherosclerosis and Renovascular Disease.</a> Circulation. 106 (9), 1165-1171 (2002).</li><li>Seddon, M., Saw, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atherosclerotic+renal+artery+stenosis:+review+of+pathophysiology,+clinical+trial+evidence,+and+management+strategies.">Atherosclerotic renal artery stenosis: review of pathophysiology, clinical trial evidence, and management strategies.</a> Can. J. Cardiol. 27 (4), 468-480 (2011).</li><li>Lao, D., Parasher, P. S., Cho, K. C., Yeghiazarians, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atherosclerotic+renal+artery+stenosis--diagnosis+and+treatment.">Atherosclerotic renal artery stenosis--diagnosis and treatment.</a> Mayo Clin Proc. 86 (7), 649-657 (2011).</li><li>Sharfuddin, A. A., Molitoris, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathophysiology+of+ischemic+acute+kidney+injury.">Pathophysiology of ischemic acute kidney injury.</a> Nat. Rev. Nephrol. 7, 189-200 (2011).</li><li>Noiri, E., 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=Oxidative+and+nitrosative+stress+in+acute+renal+ischemia.">Oxidative and nitrosative stress in acute renal ischemia.</a> Am. J. Physiol.- Renal Physiol. 281 (5), F948-F957 (2001).</li><li>Koesters, R., 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=Tubular+Overexpression+of+Transforming+Growth+Factor-&#206;1+Induces+Autophagy+and+Fibrosis+but+Not+Mesenchymal+Transition+of+Renal+Epithelial+Cells.">Tubular Overexpression of Transforming Growth Factor-&#206;1 Induces Autophagy and Fibrosis but Not Mesenchymal Transition of Renal Epithelial Cells.</a> Am. J. Pathol. 177 (2), 632-643 (2010).</li><li>Shanley, P. F., Rosen, M. D., Brezis, M., Silva, P., Epstein, F. H., Rosen, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Topography+of+focal+proximal+tubular+necrosis+after+ischemia+with+reflow+in+the+rat+kidney.">Topography of focal proximal tubular necrosis after ischemia with reflow in the rat kidney.</a> Am. J. Pathol. 122 (3), 462-468 (1986).</li></ol>

이 작품은 부분적으로 아스트라 제네카로부터 연구비에 의해 지원됩니다.

The increasing incidence of chronic kidney disease raises the need for novel therapeutic approaches that can promote functional kidney recovery7,8. Traditional therapies include the systemic administration of anti-inflammatory, anti-fibrotic drugs9. However, these strategies are frequently characterized by unwanted side effects due to off-target distribution of the injected drug. Therefore, in this manuscript, we describe a simple procedure for delivering drugs into the renal artery of rats. This pr...

The renal microvasculature is involved in a wide spectrum of kidney diseases. Depending on the pathophysiology of disease, the endothelial cells may present structural or functional impairment, which may play a pivotal role in propagating kidney damage by creating an ischemic microenvironment. This renal microvascular dysfunction may catalyze the onset of a progressive deterioration of renal function over time, leading to chronic kidney disease (CKD), end-stage renal disease, hypertension and cardiorenal syndrome. In fact, untreated hypertension may have implications in renal arterioles, causing nephrosclerosis or glomerulosclerosis with significant reduction in vascular volume fraction, increase in vascular resistance and development of tubulointerstitial fibrosis1.
Loss of renal microvasculature may be due to altered vascular homeostasis induced by local angiogenic/anti-angiogenic factors imbalance. This correlates with attenuated Vascular Endothelial Growth Factor (VEGF) signaling as well as elevated thrombospondin-12-4. Thus, using different animal models (mice, rats and pigs), the therapeutic effect of exogenous administration of VEGF has been recently investigated in some forms of renal disease, showing reduced interstitial fibrosis and stabilized renal and cardiac function3-5. This effect is likely due to actions of VEGF on endothelial cells of the microvascular bed and inflammatory monocyte phenotype switching6.
For some preclinical studies, the use of rodents, the most commonly used laboratory animals, provides a good animal model for high throughput studies due to relatively low costs and ease of handling. Moreover, the use of genetically-altered rats as models of human diseases, such as hypertension, has become more and more frequent in the scientific community. Therefore, the aim of this protocol is to describe a useful intrarenal VEGF delivery technique in rats that is easy to perform and highly reproducible. Moreover, the same method can be used to selectively deliver other drugs.

<table border="0" cellpadding="0" cellspacing="0" width="928">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Surgical Microscope</td>
			<td style="width:176px;">Leica</td>
			<td style="width:119px;">M125</td>
			<td style="width:376px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Isoflurane 100 ml</td>
			<td style="width:176px;">Cardinal Healthcare</td>
			<td style="width:119px;">PI23238</td>
			<td>Anesthetic</td>
		</tr>
		<tr height="133">
			<td height="133" style="height:133px;width:257px;">Buprenorphine HCL SR LAB 1mg/ml, 5 ml</td>
			<td style="width:176px;">ZooPharm Pharmacy</td>
			<td style="width:119px;"></td>
			<td style="width:376px;">Buprenorphine narcotic analgesic formulated in a polymer that slows absorption extending duration of action (72 hours duration of activity).&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; Liquid is viscous, warming to room temperature aids in drawing into syringe.&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; Recommended dosage: 1-1.2 mg/kg SC. DO NOT DILUTE.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:257px;">Puralube Vet Ophthalmic Ointment</td>
			<td style="width:176px;">Dechra</td>
			<td style="width:119px;">NDC17033-211-38</td>
			<td>Sterile ocular lubricant</td>
		</tr>
		<tr height="56">
			<td height="56" style="height:56px;width:257px;">Lactated Ringer&#39;s Injection, USP, 250 mL VIAFLEX Plastic Container</td>
			<td style="width:176px;">Baxter Healthcare Corp.</td>
			<td style="width:119px;">NDC0338-0117-02</td>
			<td>For body fluids replacement</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Sol Povidone-Iodine&#160; Swabstick, 3&#39;&#160;</td>
			<td style="width:176px;">Cardinal Heatlhcare</td>
			<td style="width:119px;">23405-010B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Sterile cotton tipped applicators</td>
			<td style="width:176px;">Kendall</td>
			<td style="width:119px;">8884541300</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">4-0 silk suture (without needle)&#160;</td>
			<td style="width:176px;">Cardinal Heatlhcare</td>
			<td style="width:119px;">A183H</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:257px;">Vessel Clip, Straight, 0.75 x 4mm Jaw</td>
			<td style="width:176px;">World Precision Instruments&#160;</td>
			<td style="width:119px;">501779-G</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:257px;">I.V. Catheter, Straight Hub, Radiopaque, 24g x 3/4&#34;, FEP Polymer</td>
			<td style="width:176px;">Jelco</td>
			<td align="right" style="width:119px;">4053</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Phosphate Buffered Saline</td>
			<td style="width:176px;">Life Technologies</td>
			<td align="right" style="width:119px;">10010023</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:257px;">SURGIFOAM Absorbable Gelatin Sponge</td>
			<td style="width:176px;">Cardinal Healthcare</td>
			<td align="right" style="width:119px;">179082</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:257px;">4-0 VICRYL PLUS (ANTIBACTERIAL) VIOLET 27&#34; RB-1 TAPER</td>
			<td style="width:176px;">Ethicon</td>
			<td style="width:119px;">VCP304H</td>
			<td>For muscle layer suturing</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:257px;">4-0 VICRYL PLUS (ANTIBACTERIAL) UNDYED 18&#34; PC-3 CUTTING</td>
			<td style="width:176px;">Ethicon</td>
			<td style="width:119px;">VCP845G</td>
			<td>For skin layer suturing</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:257px;">Triple antibiotic ointment</td>
			<td style="width:176px;">Actavis</td>
			<td style="width:119px;">NDC0472-0179-56</td>
			<td>For topical use on the site of the incision</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Recombinant Rat VEGF 164 Protein</td>
			<td style="width:176px;">R&#38;D Sytems</td>
			<td style="width:119px;">564-RV</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Rabbit monoclonal VEGFA</td>
			<td style="width:176px;">Abcam</td>
			<td style="width:119px;">ab46154</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Rabbit monoclonal FLK1</td>
			<td style="width:176px;">Cell Signaling</td>
			<td align="right" style="width:119px;">9698</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Rabbit monoclonal AKT</td>
			<td style="width:176px;">Cell Signaling</td>
			<td align="right" style="width:119px;">4691</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:257px;">Rabbit monoclonal phosphoAKT (Ser 473)</td>
			<td style="width:176px;">Cell Signaling</td>
			<td align="right" style="width:119px;">4060</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:257px;">Rabbit monoclonal p44/42 MAPK (ERK1/2)</td>
			<td style="width:176px;">Cell Signaling</td>
			<td align="right" style="width:119px;">4695</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:257px;">Rabbit monoclonal phospho p44/42 MAPK (Thr202 and Tyr 204)</td>
			<td style="width:176px;">Cell Signaling</td>
			<td align="right" style="width:119px;">4370</td>
			<td></td>
		</tr>
	</tbody>
</table>

intravascular delivery of biologics to the rat kidney

The renal microvascular compartment plays an important role in the progression of kidney disease and hypertension, leading to the development of End Stage Renal Disease with high risk of death for cardiovascular events. Moreover, recent clinical studies have shown that renovascular structure and function may have a great impact on functional renal recovery after surgery. Here, we describe a protocol for the delivery of drugs into the renal artery of rats. This procedure offers significant advantages over the frequently used systemic administration as it may allow a more localized therapeutic effect. In addition, the use of rodents in pharmacodynamic analysis of preclinical studies may be cost effective, paving the way for the design of translational experiments in larger animal models. Using this technique, infusion of rat recombinant Vascular Endothelial Growth Factor (VEGF) protein in rats has induced activation of VEGF signaling as shown by increased expression of FLK1, pAKT/AKT, pERK/ERK. In summary, we established a protocol for the intrarenal delivery of drugs in rats, which is simple and highly reproducible.

<html> 우리는 재조합 쥐 VEGF (rrVEGF, 0.17 μg의 / kg, 5 μg의 / kg) 또는 PBS의 두 가지 용량을 주입. 동물은 VEGF 경로의 활성을 검사 8 시간 포스트 수술을 안락사시켰다. H &amp; E 염색으로 도시 된 바와 같이, 제어부 (도 1b)에 비해 수술은 관류 신장 (도 1a)의 형태에 영향을 미치지 않았다. 제어 (그림 1D)에 비해 시리우스 레드 염색은...

Watch this Scientific Journal Video about Intravascular Delivery of Biologics to the Rat Kidney at JoVE.com

Intravascular Delivery of Biologics to the Rat Kidney

신장 기능의 회복을위한 약물의 투여는 현지화 및 치료 화합물의 분포 컨트롤이 필요합니다. 여기, 우리는 구체적으로 쥐에 약물의 신장 내 전달을위한 간단한 방법을 설명합니다. 이 절차는 쉽고없이 사망률과 높은 재현성으로 수행 될 수있다.

쥐의 신장에 생물의 혈관 배달

The renal microvascular compartment plays an important role in the progression of kidney disease and hypertension, leading to the development of End Stage ...

intravascular-delivery-of-biologics-to-the-rat-kidney

Research

JoVE Journal

Medicine

7.4K 조회수.  Mayo Clinic.  신장 기능의 회복을위한 약물의 투여는 현지화 및 치료 화합물의 분포 컨트롤이 필요합니다. 여기, 우리는 구체적으로 쥐에 약물의 신장 내 전달을위한 간단한 방법을 설명합니다. 이 절차는 쉽고없이 사망률과 높은 재현성으로 수행 될 수있다. 

비디오: Intravascular Delivery of Biologics to the Rat Kidney

Slow-release Drug Delivery through Elvax 40W to the Rat Retina: Implications for the Treatment of Chronic Conditions

Diseases of the retina are difficult to treat as the retina lies deep within the eye. Invasive methods of drug delivery are often needed to treat these diseases. Chronic retinal diseases such as retinal oedema or neovascularization usually require multiple intraocular injections to effectively treat the condition. However, the risks associated with these injections increase with repeated delivery of the drug. Therefore, alternative delivery methods need to be established in order to minimize the risks of reinjection. Several other investigations have developed methods to deliver drugs over extended time, through materials capable of releasing chemicals slowly into the eye. In this investigation, we outline the use of Elvax 40W, a copolymer resin, to act as a vehicle for drug delivery to the adult rat retina. The resin is made and loaded with the drug. The drug-resin complex is then implanted into the vitreous cavity, where it will slowly release the drug over time. This method was tested using 2-amino-4-phosphonobutyrate (APB), a glutamate analogue that blocks the light response of the retina. It was demonstrated that the APB was slowly released from the resin, and was able to block the retinal response by 7 days after implantation. This indicates that slow-release drug delivery using this copolymer resin is effective for treating the retina, and could be used therapeutically with further testing.

This paper details how Elvax 40W can be used as a slow-release method for drug delivery to the adult rat retina. The protocol for preparing, loading, and delivering the drug-resin complex to the eye is described.

쥐의 망막에 Elvax 40W를 통해 서방 약물 전달 : 만성 질환의 치료를위한 시사점

Diseases of the retina are difficult to treat as the retina lies deep within the eye. Invasive methods of drug delivery are often needed to treat these ...

연구

의학

Mouse Kidney Transplantation: Models of Allograft Rejection

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely replicates both the technical and pathological processes that occur in human renal transplantation. Although mouse models of allogeneic rejection in organs other than the kidney exist, and are more technically feasible, there is evidence that different organs elicit disparate rejection modes and dynamics, for instance the time course of rejection in cardiac and renal allograft differs significantly in certain strain combinations. This model is an attractive tool for many reasons despite its technical challenges. As inbred mouse strain haplotypes are well characterized it is possible to choose donor and recipient combinations to model acute allograft rejection by transplanting across MHC class I and II loci. Conversely by transplanting between strains with similar haplotypes a chronic process can be elicited were the allograft kidney develops interstitial fibrosis and tubular atrophy. We have modified the surgical technique to reduce operating time and improve ease of surgery, however a learning curve still needs to be overcome in order to faithfully replicate the model. This study will provide key points in the surgical procedure and aid the process of establishing this technique.

Here, we present a protocol to study the immunology of rejection. The surgical model presented reports a short operating time and a concise technique. Depending on the donor-recipient strain combination, the transplanted kidney may develop acute cellular rejection or chronic allograft damage, defined by interstitial fibrosis and tubular atrophy.

마우스 신장 이식 : 동종 이식 거부의 모델

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a serious problem. Therefore, the acceptance criteria for organ donors have been extended leading to the usage of marginal kidney grafts. These marginal organs tolerate cold storage poorly resulting in increased preservation injury and higher rates of delayed graft function. To overcome the limitations of cold storage, extensive research is focused on alternative normothermic preservation methods.
Ex vivo normothermic organ perfusion is an innovative preservation technique. The first experimental and clinical trials for ex vivo lung, liver, and kidney perfusions demonstrated favorable outcomes.
In addition to the reduction of cold ischemic injury, the method of normothermic kidney storage offers the opportunity for organ assessment and repair. This manuscript provides information about kidney retrieval, organ preservation techniques, and isolated ex vivo normothermic kidney perfusion (NEVKP) in a porcine model. Surgical techniques, set up for the perfusion solution and the circuit, potential assessment options, and representative results are demonstrated.

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

The severe organ shortage has resulted in increased use of marginal kidney grafts for transplantation. This has triggered interest in alternative storage methods, since marginal grafts especially tolerate cold storage poorly. The technique of normothermic ex vivo kidney perfusion (NEVKP) represents a novel preservation method for kidney grafts prior to &#160;transplantation.

정상 체온<em&gt; 전의 VIVO</em&gt; 신장의 보존을위한 신장 관류는 이식 전에 이식

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a ...

Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The simple murine AVF model was combined with intraluminal administration of drug solution to the venous endothelium at the same time as fistula creation. Technical aspects of this model are discussed. Under general anesthesia, an abdominal incision is made and the aorta and inferior vena cava (IVC) are exposed. The infra-renal aorta and IVC are dissected for clamping. After proximal and distal clamping, the puncture site is exposed and a 25 G needle is used to puncture both walls of the aorta and into the IVC. Immediately after the puncture, a reporter gene-expressing viral vector was infused in the IVC via the same needle, followed by 15 min of incubation. The intraluminal administration method enabled more robust viral gene delivery to the venous endothelium compared to administration by the external route. This novel method of delivery will facilitate studies that explore the role of the endothelium in AVF maturation and enable intraluminal drug delivery at the time of surgical operation.

After puncturing the aorta through the inferior vena cava (IVC) to create an aorto-caval fistula in the mouse, solution containing a drug is infused into the IVC via the same needle, followed by incubation. This method enables more robust drug delivery to the venous endothelium compared to the external route.

마우스 동정맥루 내피에 관내 약물 전달

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The ...

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ischemia/reperfusion (I/R) injury with myriad negative consequences. Potential I/R injury in marginal organs destined for liver transplantation contributes to the current donor shortage secondary to a decreased organ utilization rate. A significant need exists to explore hepatic I/R injury in order to mediate its impact on graft function in transplantation. Rat liver hilar clamp models are used to investigate the impact of different molecules on hepatic I/R injury. Depending on the model, these molecules have been delivered using inhalation, epidural infusion, intraperitoneal injection, intravenous administration or injection into the peripheral superior mesenteric vein. A rat liver hilar clamp model has been developed for use in studying the impact of pharmacologic molecules in ameliorating I/R injury. The described model for rat liver hilar clamp includes direct cannulation of the portal supply to the ischemic hepatic segment via a side branch of the portal vein, allowing for direct segmental hepatic delivery. Our approach is to induce ischemia in the left lateral and median lobes for 60 min, during which time the substance under study is infused. In this case, pegylated-superoxide dismutase (PEG-SOD), a free radical scavenger, is infused directly into the ischemic segment. This series of experiments demonstrates that infusion of PEG-SOD is protective against hepatic I/R injury. Advantages of this approach include direct injection of the molecule into the ischemic segment with consequent decrease in volume of distribution and reduction in systemic side effects.

A unique rat liver hilar clamp model was developed for studying the impact of pharmacologic molecules in ameliorating ischemia-reperfusion injury. This model includes direct cannulation of the portal supply to the ischemic liver segment via a branch of the portal vein, allowing for direct hepatic delivery.

쥐 간 문부 클램프 모델을 사용하여 직접 분절 인트라 간 납품 방법

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

Surgical Labyrinthectomy of the Rat to Study the Vestibular System

To study the vestibular system or the vestibular compensation process, a number of methods have been developed to cause vestibular damage, including surgical or chemical labyrinthectomy and vestibular neurectomy. Surgical labyrinthectomy is a relatively simple, reliable, and rapid method. Here, we describe the surgical technique for rat labyrinthectomy. A postauricular incision is made under general anesthesia to expose the external auditory canal and the tympanic membrane, after which the tympanic membrane and the ossicles are removed without the stapes. The stapes artery, which is located between the stapes and the oval window, is a vulnerable structure and must be preserved to obtain a clear surgical field. A hole to fenestrate the vestibule is made with a 2.1-mm drill bur superior to the stapes. Then, 100% ethanol is injected through this hole and aspirated several times. Meticulous dissection under a microscope and careful bleeding control are essential to obtain reliable results. Symptoms of vestibular loss, such as nystagmus, head tilting, and a rolling motion, are seen immediately after surgery. The rotarod or rotation chair test can be used to objectively and quantitatively evaluate the vestibular function.

This protocol describes the surgical labyrinthectomy of a rat, which is a useful method for studying the vestibular system.

Vestibular 시스템 공부를 쥐의 외과 Labyrinthectomy

To study the vestibular system or the vestibular compensation process, a number of methods have been developed to cause vestibular damage, including ...

A Clinical Trial Assessing the Safety, Efficacy, and Delivery of Olive-Oil-Based Three-Chamber Bags for Parenteral Nutrition

Limited evidence exists to precisely estimate efficacy and safety differences between parenteral nutrition (PN) prepared using olive-oil-based three-chamber bags (3CBs) and soybean-oil-based compounded bags (CoBs) in hospitalized adult patients. We designed a multicenter, randomized, prospective, open-label, noninferiority protocol to compare the efficacy, safety, and distribution of olive-oil-based 3CBs and soybean-oil-based CoB formulations in adult Chinese patients requiring PN during surgical intervention. Subjects were randomized to receive either one of the study treatments using an interactive voice or web-based recognition system in accordance with the randomization code. Randomization was further stratified based on the study site and surgical category. Both treatment groups received similar amounts of calories and protein. In addition, the two study treatments contained a similar composition of the amino-acid component. The only difference between the two PN formulations was the lipid constitution. The duration of administration of study treatments was a minimum of 5 days up to a maximum of 14 days after the surgical procedure. The primary efficacy endpoint was serum prealbumin levels on day 5 of the study. Noninferiority was proved if the anti-log of the lower bound of the 95% confidence interval (CI) of the treatment difference was at least 0.80. Other efficacy measures included treatment preparation time; duration to achieve tolerability of oral nutrition; associated infectious complications; length of hospitalization; and laboratory assessment of markers of nutrition, inflammation, metabolism, and oxidative stress. A total of 458 patients were enrolled in the study. The results showed that olive-oil-based 3CBs were non-inferior to soybean-based CoBs, besides being well tolerated. The infection rate was found to be significantly lower in the olive-oil-based 3CB group. Thus, this study may be used as a reference for future research on lipid emulsion and 3CBs.

Here, we present a protocol for a study comparing the efficacy, safety, and delivery of olive-oil-based 3CB and soybean-oil-based CoB formulations in adults requiring parenteral nutrition. The results revealed that olive-oil-based 3CBs is non-inferior and well tolerated compared to soybean formulations.

Limited evidence exists to precisely estimate efficacy and safety differences between parenteral nutrition (PN) prepared using olive-oil-based ...

An Efficient Sieving Method to Isolate Intact Glomeruli from Adult Rat Kidney

Preservation of glomerular structure and function is pivotal in the prevention of glomerulonephritis, a category of kidney disease characterized by proteinuria which can eventually lead to chronic and end-stage renal disease. The glomerulus is a complex apparatus responsible for the filtration of plasma from the body. In disease, structural integrity is lost and allows for the abnormal leakage of plasma contents into the urine. A method to isolate and examine glomeruli in culture is critical for the study of these diseases. In this protocol, an efficient method of retrieving intact glomeruli from adult rat kidneys while conserving structural and morphological characteristics is described. This process is capable of generating high yields of glomeruli per kidney with minimal contamination from other nephron segments. With these glomeruli, injury conditions can be mimicked by incubating them with a variety of chemical toxins, including protamine sulfate, which causes foot process effacement and proteinuria in animal models. Degree of injury can be assessed using transmission electron microscopy, immunofluorescence staining, and western blotting. Nephrin and Wilms Tumor 1 (WT1) levels can also be assessed from these cultures. Due to the ease and flexibility of this protocol, the isolated glomeruli can be utilized as described or in a way that best suits the needs of the researcher to help better study glomerular health and structure in diseased states.

The main focus of this protocol is to efficiently isolate viable primary glomeruli cultures with minimal contaminants for use in a variety of downstream applications. The isolated glomeruli retain structural relationships between component cell types and can be cultured ex vivo for a short time.

그대로 Glomeruli 성인 쥐 신장에서 분리 하는 효율적인 체질 하 방법

Preservation of glomerular structure and function is pivotal in the prevention of glomerulonephritis, a category of kidney disease characterized by ...

Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of renal replacement therapy. Over the past few decades, the rat kidney transplantation model has been used to study the immunological phenomena of rejection and tolerance. This model has become an indispensable tool to test new immunomodulatory pharmaceuticals and regimens prior to proceeding with expensive preclinical large animal studies.
This protocol provides a detailed overview of how to reliably perform orthotopic kidney transplantation in rats. This protocol includes three distinctive steps that increase the probability of success: perfusion of the donor kidney by flushing through the portal vein and the use of a cuff system to anastomose the renal veins and ureters, thereby decreasing cold and warm ischemia times. Using this technique, we have achieved survival rates beyond 6 months with normal serum creatinine in animals with syngeneic or tolerant kidney transplants. Depending on the aim of the study, this model can be modified by pre- or posttransplant treatments to study the acute, chronic, cellular, or antibody-mediated rejection. It is a reproducible, reliable, and cost-effective animal model to study different aspects of kidney transplantation.

The purpose of this manuscript and protocol is to explain and demonstrate in detail the surgical procedure of orthotopic kidney transplantation in rats. This method is simplified to achieve the correct perfusion of the donor kidney and shorten the reperfusion time by using the venous and ureteral cuff anastomosis technique.

직교 쥐 신장 이식: 새롭고 단순화 된 외과 접근

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of ...

Intratracheal Administration of Dry Powder Formulation in Mice

In the development of inhalable dry powder formulations, it is essential to evaluate their biological activities in preclinical animal models. This paper introduces a noninvasive method of intratracheal delivery of dry powder formulation in mice. A dry powder loading device that consists of a 200 &#181;L gel loading pipette tip connected to an 1 mL syringe via a three-way stopcock is presented. A small amount of dry powder (1-2 mg) is loaded into the pipette tip and dispersed by 0.6 mL of air in the syringe. Because pipette tips are disposable and inexpensive, different dry powder formulations can be loaded into different tips in advance. Various formulations can be evaluated in the same animal experiment without device cleaning and dose refilling, thereby saving time and eliminating the risk of cross-contamination from residual powder. The extent of powder dispersion can be inspected by the amount of powder remaining in the pipette tip. A protocol of intubation in mouse with a custom-made light source and a guiding cannula is included. Proper intubation is one of the key factors that influences the intratracheal delivery of dry powder formulation to the deep lung region of the mouse.

Dry powder formulations for inhalation have great potential in treating respiratory diseases. Before entering human studies, it is necessary to evaluate the efficacy of the dry powder formulation in preclinical studies. A simple and noninvasive method of the administration of dry powder in mice through the intratracheal route is presented.

쥐에서 건조 분말 제형의 장내 투여

In the development of inhalable dry powder formulations, it is essential to evaluate their biological activities in preclinical animal models. This paper ...