本文選題：絲氨酸羥甲基轉(zhuǎn)移酶2 + 肝再生�。� 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:探索絲氨酸羥甲基轉(zhuǎn)移酶2(serine hydroxymethyl transferase 2,SHMT2)在小鼠肝再生過程中的表達(dá)規(guī)律,以及其在小鼠肝再生過程中發(fā)揮的作用。方法:1.SPF級(jí)C57BL/6小鼠90只分為肝切除組(2/3PH)50只、假手術(shù)組(Sham組)10只、尾靜脈注射攜帶SHMT2 sh RNA腺病毒組(sh SHMT2組)10只、尾靜脈注射陰性對(duì)照腺病毒組(Control組)10只、尾靜脈注射生理鹽水空白對(duì)照組(NS組)10只。小鼠肝切除之后,記錄切除肝臟的重量,并在對(duì)應(yīng)的時(shí)間點(diǎn)1、3、5、7、9 d收集肝臟組織及血清。同時(shí)收集Sham組的肝臟組織及血清,用qPCR、Western blot(WB)和免疫組化檢測(cè)SHMT2及其下游分子甘氨酸脫羧酶(glycine decarboxylase,GLDC)的變化規(guī)律,自動(dòng)生化分析儀檢測(cè)血清中丙氨酸轉(zhuǎn)氨酶(alanine aminotransferase,ALT)和谷草轉(zhuǎn)氨酶(aspartate transaminase,AST)的水平。2.取shSHMT2組、Control組及NS組的小鼠肝臟組織,冰凍切片觀察綠色熒光強(qiáng)度,Western blot檢測(cè)SHMT2表達(dá)量驗(yàn)證腺病毒的干擾效率,用免疫組化法檢測(cè)三組肝臟組織中增殖細(xì)胞核抗原(Proliferating Cell Nuclear Antigen,PCNA)的表達(dá),比較三者肝再肝臟重量、肝再生度、肝指數(shù)及小鼠血清中的ALT、AST水平。結(jié)果:SHMT2及其下游分子GLDC,均在2/3肝切除術(shù)后開始升高,并分別在小鼠肝再生的后第5天,第7天達(dá)到峰值,qPCR結(jié)果揭示了與WB相似的趨勢(shì),SHMT2 m RNA表達(dá)量逐漸上升在第5天達(dá)到最高為第1天的1.63倍(P0.05)。在干擾SHMT2之后,NS組與sh SHMT2組比較,再生肝臟的重量由(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)%,肝指數(shù)由(3.76±0.44)%降低至(3.13±0.29)%,并且肝功能指標(biāo)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陽(yáng)性率從(53.6±2.3)%降低至(39.0±3.2)%(P0.05)。結(jié)論:1.SHMT2在肝臟再生過程中表達(dá)水平有明顯變化,在PH之后開始上升,且在第5天達(dá)到峰值。2.SHMT2表達(dá)被干擾后,小鼠肝再生度及肝功能均有所降低,肝臟增殖減弱,說明SHMT2對(duì)肝臟再生有促進(jìn)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緇勪笌sh SHMT2緇勬瘮杈，

本文編號(hào)：2075900