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目的通过动物在体实验了解组织型谷氨酰胺转移酶(tissue transglutaminase,tTG)抑制剂(monodansyl-cadaverine,MDC)对凝血酶(thrombin,Tm)脑组织毒性的影响。方法第1组动物用4种剂量的Tm(0、5、10、15U溶于5μl生理盐水)通过立体定向仪注射入大鼠纹状体,第2组动物在各剂量Tm中同时加入MDC(5mM)加以干预作为相应对照,动物存活24h后取注射部位石蜡包埋切片,做常规HE染色和凋亡特异性TUNEL染色及照相。观察两种染色的切片细胞变化并对各组TUNEL特异染色的凋亡细胞进行计数,以了解在大鼠体内MDC抑制tTG对Tm诱导凋亡作用的影响。结果Tm处理脑组织的胞体发生肿胀,细胞核发生浓缩固缩现象,核仁消失,核膜完整性破坏,且浓度越高异常形态的细胞越多。而在对照的Tm+MDC处理组织切片的异常细胞减少。TUNEL凋亡细胞特异性染色计数,5、10和15UTm凋亡细胞数分别为(13.78±3.31)、(25.56±4.16)和(35.00±5.96)个;而同时加入MDC3种不同浓度的凋亡细胞数分别为(8.56±2.51)、(16.89±2.03)和(28.33±5.83)个。结论MDC可通过抑制tTG减少Tm引起的细胞凋亡。
OBJECTIVE: To understand the effects of tissue transglutaminase (tTG) inhibitor (montansyl-cadaverine) on thrombin (Tm) brain tissue toxicity in vivo. Methods Group 1 animals were injected into rat striatum with 4 doses of Tm (0, 5, 10, 15U dissolved in 5 μl saline) by stereotactic apparatus and Group 2 animals were dosed with MDC at each dose of Tm 5mM) as the corresponding control. After the animals survived for 24 hours, the injection site was paraffin-embedded, and routine HE staining and apoptosis-specific TUNEL staining were performed. The changes of the two stained sections were observed and the number of TUNEL-stained apoptotic cells in each group was counted to find out the effect of MDC inhibition of tTG on Tm-induced apoptosis in rats. Results Tm treatment of brain tissue swelling, nuclear condensation phenomenon, nucleolus disappear, nuclear integrity damage, and the higher the concentration of abnormal morphology of the cells. Whereas abnormal cell depletion in the control Tm + MDC treated tissue sections. The number of apoptotic cells at 5, 10 and 15UTm was (13.78 ± 3.31), (25.56 ± 4.16) and (35.00 ± 5.96), respectively, while the apoptotic cells The numbers were (8.56 ± 2.51), (16.89 ± 2.03) and (28.33 ± 5.83), respectively. Conclusion MDC can reduce Tm-induced apoptosis by inhibiting tTG.