丝氨酸羟甲基转移酶2在小鼠肝再生过程中的变化规律及作用
发布时间：2018-11-06 12:30
【摘要】：目的探索丝氨酸羟甲基转移酶2(serine hydroxymethyl transferase 2,SHMT2)在小鼠肝再生过程中的表达规律,以及其对小鼠肝再生是否有促进作用。方法 SPF级C57BL/6小鼠120只分为肝切除组(2/3PH)、假手术组(SHAM)、腺病毒干扰组(si SHMT2)及注射生理盐水对照组(Control),每组30只;肝切除组下设1、3、5、7、9 d 5个时相点,每个时相点6只。小鼠肝切除之后分别取1、3、5、7、9 d的肝脏组织及血清,采用q PCR、Western blot和免疫组化检测SHMT2及其下游分子甘氨酸脱氢酶(glycine dehydrogenase,GLDC)的变化规律、血清中丙氨酸转氨酶(alanine transaminase,ALT)和谷草转氨酶(aspartate transaminase,AST)的浓度。取注射腺病毒干扰SHMT2第5天的肝脏组织,通过Western blot和观察荧光强度,确定转染效果,并检测血浆中ALT和AST水平,免疫组化检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)。结果 SHMT2及其下游分子GLDC在小鼠肝再生的后第5、7天表达最高,q PCR检测结果显示,SHMT2第5天的表达量是第1天的1.63倍(P0.05)。在干扰SHMT2之后,小鼠第5天再生肝脏的质量由(0.79±0.13)g降低至(0.63±0.11)g(P0.05),再生度由(36.37±2.21)%降低至(31.33±1.92)%,肝指数由(3.76±0.44)%降低至(3.13±0.29)%,并且肝功能指标ALT由(70.00±9.52)U/L升高至(154.15±16.49)U/L,AST由(140.09±32.85)U/L升高至(403.41±68.63)U/L(P0.05),PCNA阳性率从(53.6±2.3)%降低至(39.0±3.2)%(P0.05)。结论 SHMT2在肝脏再生的后期高表达,促进残肝的再生,其机制可能与增强肝脏对缺血、缺氧的耐受,提高三磷酸腺苷水平有关。
[Abstract]:Objective to investigate the expression of serine hydroxymethyltransferase 2 (serine hydroxymethyl transferase 2 (SHMT2) in mouse liver regeneration and whether it can promote liver regeneration in mice. Methods 120 SPF C57BL/6 mice were divided into hepatectomy group (2/3PH), (SHAM), adenovirus interference group (si SHMT2) and saline injection control group (Control),). In the hepatectomy group, there were 5 time points on 9 days, 6 in each. After hepatectomy, liver tissues and serum from mice were collected for 9 days. The changes of SHMT2 and its downstream molecule glycine dehydrogenase (glycine dehydrogenase,GLDC) were detected by Q PCR,Western blot and immunohistochemistry. The alanine aminotransferase (alanine transaminase,) in serum was also detected. Concentrations of ALT and aspartate transaminase,AST. The liver tissue which was interfered by adenovirus on the 5th day of SHMT2 was taken. The transfection effect was determined by Western blot and fluorescence intensity. The levels of ALT and AST in plasma and proliferating cell nuclear antigen (proliferating cell nuclear antigen,PCNA) were detected by immunohistochemistry. Results the highest expression of SHMT2 and its downstream GLDC was detected on the 5th day after liver regeneration in mice. The results showed that the expression of SHMT2 on the 5th day was 1.63 times of that on the first day (P0.05). After interfering with SHMT2, the weight of regenerated liver decreased from (0.79 卤0.13) g to (0.63 卤0.11) g (), the degree of regeneration decreased from (36.37 卤2.21)% to (31.33 卤1.92)%. The liver index decreased from (3.76 卤0.44)% to (3.13 卤0.29)%, and the liver function index (ALT) increased from (70.00 卤9.52) U / L to (154.15 卤16.49) U / L, respectively. AST increased from (140.09 卤32.85) U / L to (403.41 卤68.63) U / L (P0.05), PCNA positive rate decreased from (53.6 卤2.3)% to (39.0 卤3.2)% (P0.05). Conclusion SHMT2 is highly expressed in the late stage of liver regeneration and promotes the regeneration of residual liver. The mechanism may be related to the enhancement of liver tolerance to ischemia and hypoxia and the increase of adenosine triphosphate level.
【作者单位】： 重庆医科大学第二附属医院肝胆外科;
【分类号】：R657.3
[Abstract]:Objective to investigate the expression of serine hydroxymethyltransferase 2 (serine hydroxymethyl transferase 2 (SHMT2) in mouse liver regeneration and whether it can promote liver regeneration in mice. Methods 120 SPF C57BL/6 mice were divided into hepatectomy group (2/3PH), (SHAM), adenovirus interference group (si SHMT2) and saline injection control group (Control),). In the hepatectomy group, there were 5 time points on 9 days, 6 in each. After hepatectomy, liver tissues and serum from mice were collected for 9 days. The changes of SHMT2 and its downstream molecule glycine dehydrogenase (glycine dehydrogenase,GLDC) were detected by Q PCR,Western blot and immunohistochemistry. The alanine aminotransferase (alanine transaminase,) in serum was also detected. Concentrations of ALT and aspartate transaminase,AST. The liver tissue which was interfered by adenovirus on the 5th day of SHMT2 was taken. The transfection effect was determined by Western blot and fluorescence intensity. The levels of ALT and AST in plasma and proliferating cell nuclear antigen (proliferating cell nuclear antigen,PCNA) were detected by immunohistochemistry. Results the highest expression of SHMT2 and its downstream GLDC was detected on the 5th day after liver regeneration in mice. The results showed that the expression of SHMT2 on the 5th day was 1.63 times of that on the first day (P0.05). After interfering with SHMT2, the weight of regenerated liver decreased from (0.79 卤0.13) g to (0.63 卤0.11) g (), the degree of regeneration decreased from (36.37 卤2.21)% to (31.33 卤1.92)%. The liver index decreased from (3.76 卤0.44)% to (3.13 卤0.29)%, and the liver function index (ALT) increased from (70.00 卤9.52) U / L to (154.15 卤16.49) U / L, respectively. AST increased from (140.09 卤32.85) U / L to (403.41 卤68.63) U / L (P0.05), PCNA positive rate decreased from (53.6 卤2.3)% to (39.0 卤3.2)% (P0.05). Conclusion SHMT2 is highly expressed in the late stage of liver regeneration and promotes the regeneration of residual liver. The mechanism may be related to the enhancement of liver tolerance to ischemia and hypoxia and the increase of adenosine triphosphate level.
【作者单位】： 重庆医科大学第二附属医院肝胆外科;
【分类号】：R657.3
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