【摘要】：目的建立山羊擠壓綜合征(CS)動(dòng)物模型,并探討連續(xù)性靜脈-靜脈血液濾過(CVVH)治療對(duì)腎功能及腎組織病理變化的影響。方法 12只山羊隨機(jī)分為對(duì)照組、CS模型組、CVVH治療組,每組4只。CS模型組與CVVH治療組山羊于后肢肌肉注射50%甘油生理鹽水溶液10 mL/kg建立CS模型(對(duì)照組注射等量生理鹽水),1、2、8、12、24h后檢測(cè)血清肌酸激酶(sCK)和血肌酐(sCr),以sCK1 000U/L,同時(shí)sCr2倍對(duì)照值判斷為造模成功。造模1h后,CVVH治療組于造模對(duì)側(cè)股靜脈留置單針雙腔導(dǎo)管建立血管通路,采用智能化床旁腎臟替代治療機(jī)進(jìn)行CVVH治療。血流量為100mL/min,置換液流速為35mL/(kg·h),以前稀釋法輸入,治療23h后(第24h)處死動(dòng)物,留取腎組織標(biāo)本,光鏡、電鏡下觀察腎組織病理組織結(jié)構(gòu)變化;采用免疫組化染色檢查caspase12蛋白表達(dá)變化及TUNEL染色對(duì)細(xì)胞凋亡進(jìn)行檢測(cè)。結(jié)果造模山羊均于注射甘油后1h內(nèi)出現(xiàn)醬油色小便,尿量較對(duì)照組明顯減少。CS模型組與CVVH治療組造模1h后血清sCr及sCK較對(duì)照組升高,差異有統(tǒng)計(jì)學(xué)意義(P0.05),提示造模成功。行CVVH治療23h(第24h),sCK、sCr水平低于CS模型組(P0.05)。光鏡下CS模型組腎組織可見急性腎小管壞死表現(xiàn),腎間質(zhì)水腫,腎小球基本正常。電鏡下模型組可見小管上皮細(xì)胞明顯的染色質(zhì)聚集,線粒體腫脹,內(nèi)質(zhì)網(wǎng)擴(kuò)張等早期細(xì)胞凋亡征象,CVVH治療組表現(xiàn)稍輕。CS模型組及CVVH治療組caspase12表達(dá)高于對(duì)照組(P0.001),CVVH治療組caspase12表達(dá)低于CS模型組(P0.05)。TUNEL染色證實(shí)CS模型組山羊腎組織細(xì)胞凋亡比例明顯增高,CVVH治療組較模型組凋亡比例少。結(jié)論通過肌肉注射50%甘油生理鹽水溶液可建立山羊CS模型,早期行CVVH治療可減緩腎功能惡化,減輕腎小管上皮細(xì)胞凋亡程度。
[Abstract]:Objective To establish an animal model of goat crush syndrome (CS) and to explore the effect of continuous veno-venous hemofiltration (CVVH) on the pathological changes of renal and renal tissues. Methods 12 goats were randomly divided into control group, CS model group and CVVH group. CS model group and CVVH treatment group were used to establish CS model (control group to inject the same amount of normal saline),1,2,8,12 and 24 hours, and the serum creatine kinase (sCK) and blood myoglobin (sCr) were detected, and the sCK1 000 U/ L and sCr2 times of the control value were determined as the success of the model. After the model was set for 1 h, the CVVH treatment group was used to set up the vascular access with a single-needle double-lumen catheter in the contralateral femoral vein of the model, and the CVVH was treated with an intelligent bedside renal replacement therapy machine. The blood flow was 100 mL/ min, the flow rate of the replacement solution was 35 mL/ (kg 路 h), and the blood flow was input by the previous dilution method. After the treatment for 23 h (day 24), the animal was sacrificed, and the renal tissue specimen, the light microscope and the electron microscope were used to observe the structural changes of the renal tissue. The expression of caspase-12 and the TUNEL staining were used to detect the apoptosis of the cells. Results The amount of urine in soy sauce appeared in the model goat after injection of glycerol for 1 h, and the urine volume was significantly reduced in the control group. The serum sCr and sCK in the CS model group and the CVVH group were higher than that in the control group, and the difference was significant (P0.05). The levels of sCK and sCr were lower than that of CS model group (P0.05). In the light microscope, the renal tissue of the CS model group showed acute tubular necrosis, renal interstitial edema, and normal glomerulus. In the model group under the electron microscope, the apoptosis of the early cells such as the aggregation of chromatin, the swelling of mitochondria and the expansion of the endoplasmic reticulum were observed. The expression of caspase12 in the CS model group and the CVVH group was higher than that in the control group (P0.001), and the expression of caspase12 in the CVVH group was lower than that in the CS model group (P0.05). Conclusion The model of goat CS can be established by intramuscular injection of 50% glycerin physiological saline solution, and the early stage CVVH treatment can slow the deterioration of renal function and reduce the apoptosis of renal tubular epithelial cells.
【作者單位】： 四川大學(xué)華西醫(yī)院腎臟內(nèi)科;
【基金】：國家自然科學(xué)基金(No.81270818)資助
【分類號(hào)】：R642;R-332

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