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目的 探讨TaT融合蛋白在SD大鼠心脏组织的吸收及酶活性的表达情况。方法 利用“蛋白转导”的方法 ,将源于人类免疫缺陷蛋白病毒HIV的TaT蛋白与 β -半乳糖苷酶 (β -galactosidase ,β -Gal)相连接 ,构建成分子量 12 0KD的TaT -β -Gal融合蛋白。随机将 72只SD大鼠分为 3组 (n =2 4)分别腹腔注射TaT -β -Gal、β -Gal及生理盐水 ,并于注射后不同时间段取心脏病理切片 ,用X -Gal染色分析 ,显微镜下观察心脏组织对TaT -β -Gal的吸收及酶活性的表达情况 ,并进行Leica计算机图象分析 ,同时观察心脏组织结构变化。结果 经X -Gal染色后 ,TaT -β -Gal组 0 5h显微镜下心脏组织即显示着色反应 ,表达酶活性 ,8h达到高峰 ,16h后逐渐下降 ,心脏组织结构无变化 ;β -Gal组出现极弱的着色反映 ;生理盐水组无着色反应。 结论 分子量 12 0KD的TaT-β -Gal融合蛋白可以迅速被心脏组织吸收 ,同时不影响酶活性。TaT蛋白可作为载体蛋白转运大分子物质通过生物膜进入心肌细胞。
Objective To investigate the expression of TaT fusion protein in SD rat heart tissue. Methods TaT protein derived from human immunodeficiency virus HIV was linked to β-galactosidase (β -Gal) by using “protein transduction” method to construct TaT-β with molecular weight of 120KD -Gal fusion protein. 72 SD rats were randomly divided into 3 groups (n = 24). TaT-β-Gal, β-Gal and normal saline were injected intraperitoneally. Cardiac pathological sections were taken at different time points after injection, and stained with X-Gal The expression of TaT-β-Gal in cardiac tissue was observed under a microscope, and the Leica computer image analysis was performed. Meanwhile, the changes of cardiac tissue structure were also observed. Results After staining with X-Gal, the histopathology of the TaT-β-Gal group at 0h showed a color reaction, which reached the peak at 8h and then decreased gradually after 16h, and there was no change in cardiac structure. Weak coloring reaction; no staining reaction in saline group. Conclusion TaT-β-Gal fusion protein with a molecular weight of 120KD can be quickly absorbed by cardiac tissues without affecting the enzyme activity. TaT protein as a carrier protein transport macromolecules through the biofilm into cardiomyocytes.