SHMT2在小鼠肝再生过程中的变化规律及作用

发布时间：2018-06-28 00:31

  本文选题：丝氨酸羟甲基转移酶2 + 肝再生　； 参考：《重庆医科大学》2017年硕士论文

【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,以及其在小鼠肝再生过程中发挥的作用。方法:1.SPF级C57BL/6小鼠90只分为肝切除组(2/3PH)50只、假手术组(Sham组)10只、尾静脉注射携带SHMT2 sh RNA腺病毒组(sh SHMT2组)10只、尾静脉注射阴性对照腺病毒组(Control组)10只、尾静脉注射生理盐水空白对照组(NS组)10只。小鼠肝切除之后,记录切除肝脏的重量,并在对应的时间点1、3、5、7、9 d收集肝脏组织及血清。同时收集Sham组的肝脏组织及血清,用qPCR、Western blot(WB)和免疫组化检测SHMT2及其下游分子甘氨酸脱羧酶(glycine decarboxylase,GLDC)的变化规律,自动生化分析仪检测血清中丙氨酸转氨酶(alanine aminotransferase,ALT)和谷草转氨酶(aspartate transaminase,AST)的水平。2.取shSHMT2组、Control组及NS组的小鼠肝脏组织,冰冻切片观察绿色荧光强度,Western blot检测SHMT2表达量验证腺病毒的干扰效率,用免疫组化法检测三组肝脏组织中增殖细胞核抗原(Proliferating Cell Nuclear Antigen,PCNA)的表达,比较三者肝再肝脏重量、肝再生度、肝指数及小鼠血清中的ALT、AST水平。结果:SHMT2及其下游分子GLDC,均在2/3肝切除术后开始升高,并分别在小鼠肝再生的后第5天,第7天达到峰值,qPCR结果揭示了与WB相似的趋势,SHMT2 m RNA表达量逐渐上升在第5天达到最高为第1天的1.63倍(P0.05)。在干扰SHMT2之后,NS组与sh SHMT2组比较,再生肝脏的重量由(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:1.SHMT2在肝脏再生过程中表达水平有明显变化,在PH之后开始上升,且在第5天达到峰值。2.SHMT2表达被干扰后,小鼠肝再生度及肝功能均有所降低,肝脏增殖减弱,说明SHMT2对肝脏再生有促进作用。
[Abstract]:Aim: to investigate the expression of serine hydroxymethyltransferase 2 (serine hydroxymethyl transferase 2 (SHMT2) in mouse liver regeneration and its role in mouse liver regeneration. Methods 90 SPF C57BL / 6 mice were divided into hepatectomy group (n = 50), sham-operated group (n = 10), adenovirus group with SHMT2sh RNA (n = 10) and negative control group (control group) (n = 10). Ten rats in the control group (NS group) were injected with normal saline via caudal vein. After hepatectomy, the weight of the liver was recorded, and the liver tissue and serum were collected at the corresponding time point of 1? The liver tissue and serum of Sham group were collected at the same time. The changes of SHMT2 and its downstream molecule glycine decarboxylase GLDC were detected by qPCRX Western blot (WB) and immunohistochemistry. The serum levels of alanine aminotransferase (alt) and aspartate transaminase (aspartate) were detected by automatic biochemical analyzer. The liver tissues of control group and NS group were taken from shSHMT2 group. The expression of SHMT2 was detected by Western blot with green fluorescence intensity and the expression of proliferating cell nuclear antigen (PCNA) was detected by immunohistochemical method. Liver weight, liver regeneration, liver index and serum alt AST were compared among the three groups. Results both the fraction SHMT2 and its downstream GLDC2 began to increase after 2 / 3 hepatectomy, and were observed on the fifth day after liver regeneration in mice. The peak value of QPCR on the 7th day revealed that the expression of SHMT2 mRNA gradually increased to 1.63 times of that on the fifth day (P0.05). 鍦ㄥ共鎵癝HMT2涔嬪悗,NS缁勪笌sh SHMT2缁勬瘮杈，

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