發(fā)布時(shí)間：2018-03-29 05:09

  本文選題：去細(xì)胞　切入點(diǎn)：腎臟　出處：《解剖學(xué)報(bào)》2014年02期

【摘要】：目的通過在體灌注Triton X-100、SDS溶液法制備大鼠全腎去細(xì)胞生物支架,并對(duì)支架制備過程中的關(guān)鍵步驟進(jìn)行程序性檢測、分析,為制備科學(xué)合理的實(shí)驗(yàn)用大鼠全腎去細(xì)胞生物支架提供基礎(chǔ)。方法 SD大鼠40只,隨機(jī)分為4組,每組10只。肝素化后直接切取腎臟作為對(duì)照組(control組);肝素PBS灌注組(H組);肝素PBS、Triton X-100灌注組(HT組);肝素PBS、Triton X-100、十二烷基硫酸鈉(SDS)溶液灌注組(HTS組)。HE、Masson、PAS染色及透射電鏡觀察各組腎組織病理及超微結(jié)構(gòu)改變,免疫熒光結(jié)合4,6-二脒基-2-苯基吲哚(DAPI)觀察各組膠原蛋白Ⅳ(collagenⅣ)、層黏連蛋白(LN)、纖維連接蛋白(FN)、硫酸乙酰肝素蛋白多糖2(HSPG2)、彈性蛋白(elastin)及細(xì)胞核的表達(dá)情況,總DNA測定各組DNA殘留濃度。結(jié)果 Triton X-100、SDS灌注6h左右制備大鼠腎臟去細(xì)胞生物支架。在灌注過程中,腎內(nèi)細(xì)胞和細(xì)胞碎片逐漸被清洗,最終變成半透明狀。HE、Masson、PAS染色及透射電鏡顯示連續(xù)分布、形態(tài)排列類似腎小球、腎小管輪廓結(jié)構(gòu)的網(wǎng)狀結(jié)構(gòu),基膜連續(xù)完整。免疫熒光結(jié)合DAPI染色結(jié)果表明,去細(xì)胞過程中細(xì)胞外基質(zhì)中的重要蛋白collagen IV、LN、FN、HSPG2、elastin都較好的得到了保存,細(xì)胞核在灌注過程中逐漸減少,直至完全消失;最終殘留組織的DNA為4.90μg/g。結(jié)論通過合適濃度和灌注比例的Triton X-100、SDS制備大鼠腎臟去細(xì)胞生物支架,并對(duì)其進(jìn)行程序性分析,發(fā)現(xiàn)能有效清除大鼠腎內(nèi)所有細(xì)胞成分,較完整的保留網(wǎng)絡(luò)狀腎及腎血管細(xì)胞外基質(zhì)結(jié)構(gòu)和成分,是一種簡單易行且較為理想的制備實(shí)驗(yàn)用全腎生物支架的方法。
[Abstract]:Objective to prepare rat renal acellular scaffolds by in vivo perfusion of Triton X-100 SDS solution, and to detect and analyze the key steps in the process of scaffold preparation. Methods Forty SD rats were randomly divided into 4 groups. 10 rats in each group. After heparinization, the kidneys were directly removed as control group; heparin PBS perfusion group was treated with H group; heparin PBS group was perfused with Triton X-100; heparin PBSI Triton X-100, dodecyl sulfate sodium sulfate solution was perfused with HES group; heparin PBS group with Triton Triton X-100; heparin #en1# group with Triton Triton pas staining and transmission electron microscopy (TEM). The pathological and ultrastructural changes of renal tissue were observed in each group. The expression of collagen 鈪，

本文編號(hào)：1679676