南通大学学报（医学版）
南通大學學報（醫學版）
남통대학학보（의학판）
JOURNAL OF NANTONG UNIVERSITY(MEDICAL SCIENCES)
2015年
2期
102-105
,共4页
邵晓%何红梅%邵建国%陈琳%李民
邵曉%何紅梅%邵建國%陳琳%李民
소효%하홍매%소건국%진림%리민
Hck%Lyn%Src抑制的蛋白激酶C底物%Kupffer细胞%炎症
Hck%Lyn%Src抑製的蛋白激酶C底物%Kupffer細胞%炎癥
Hck%Lyn%Src억제적단백격매C저물%Kupffer세포%염증
Hck%Lyn%Src-suppressed C kinase substrate%Kupffer cell%inflammation
目的：探讨Src家族相关激酶（Hck、Lyn）和Src抑制的蛋白激酶C底物（Src-suppressed C kinase substrate， SSeCKS）在Kupffer细胞中的表达及意义。方法：应用 Real-time PCR的方法检测正常Kupffer细胞和炎症Kupffer细胞中Hck、Lyn和SSeCKS的表达水平；用Western Blot的方法检测Hck、Lyn和SSeCKS在Kupffer细胞中的表达水平。构建非酒精性脂肪性肝炎（non-alcoholic steatohepatitis, NASH）大鼠模型，采用免疫组织化学（immunohistochemistry, IHC）方法检测Hck、Lyn和SSeCKS在大鼠肝脏组织Kupffer细胞中的表达水平。结果：RT-PCR显示Hck、Lyn和SSeCKS在小鼠Kupffer细胞中均有表达，用60%CCl4损伤液刺激后Hck、Lyn表达明显升高(P<0.01)，SSeCKS表达减少，但差异无统计学意义(P＞0.05)；Western Blot结果显示在Kupffer细胞中Hck、SSeCKS表达较为明显，Lyn未见明显表达；IHC结果显示Hck、Lyn和SSeCKS在大鼠正常肝组织Kupffer中均有表达；在炎症组织中Hck、Lyn的表达明显增加，而SSeCKS的表达却显著减少。在炎症刺激后Hck、Lyn的表达与Kupffer细胞数呈正相关，而SSeCKS的表达与Kupffer细胞数呈负相关。结论：Hck、Lyn和SSeCKS参与了NASH的发生发展，可作为NASH炎症反应的潜在调控靶点；检测其表达有助于NASH炎症程度的判断，为临床上治疗NASH提供一定的客观依据。
目的：探討Src傢族相關激酶（Hck、Lyn）和Src抑製的蛋白激酶C底物（Src-suppressed C kinase substrate， SSeCKS）在Kupffer細胞中的錶達及意義。方法：應用 Real-time PCR的方法檢測正常Kupffer細胞和炎癥Kupffer細胞中Hck、Lyn和SSeCKS的錶達水平；用Western Blot的方法檢測Hck、Lyn和SSeCKS在Kupffer細胞中的錶達水平。構建非酒精性脂肪性肝炎（non-alcoholic steatohepatitis, NASH）大鼠模型，採用免疫組織化學（immunohistochemistry, IHC）方法檢測Hck、Lyn和SSeCKS在大鼠肝髒組織Kupffer細胞中的錶達水平。結果：RT-PCR顯示Hck、Lyn和SSeCKS在小鼠Kupffer細胞中均有錶達，用60%CCl4損傷液刺激後Hck、Lyn錶達明顯升高(P<0.01)，SSeCKS錶達減少，但差異無統計學意義(P＞0.05)；Western Blot結果顯示在Kupffer細胞中Hck、SSeCKS錶達較為明顯，Lyn未見明顯錶達；IHC結果顯示Hck、Lyn和SSeCKS在大鼠正常肝組織Kupffer中均有錶達；在炎癥組織中Hck、Lyn的錶達明顯增加，而SSeCKS的錶達卻顯著減少。在炎癥刺激後Hck、Lyn的錶達與Kupffer細胞數呈正相關，而SSeCKS的錶達與Kupffer細胞數呈負相關。結論：Hck、Lyn和SSeCKS參與瞭NASH的髮生髮展，可作為NASH炎癥反應的潛在調控靶點；檢測其錶達有助于NASH炎癥程度的判斷，為臨床上治療NASH提供一定的客觀依據。
목적：탐토Src가족상관격매（Hck、Lyn）화Src억제적단백격매C저물（Src-suppressed C kinase substrate， SSeCKS）재Kupffer세포중적표체급의의。방법：응용 Real-time PCR적방법검측정상Kupffer세포화염증Kupffer세포중Hck、Lyn화SSeCKS적표체수평；용Western Blot적방법검측Hck、Lyn화SSeCKS재Kupffer세포중적표체수평。구건비주정성지방성간염（non-alcoholic steatohepatitis, NASH）대서모형，채용면역조직화학（immunohistochemistry, IHC）방법검측Hck、Lyn화SSeCKS재대서간장조직Kupffer세포중적표체수평。결과：RT-PCR현시Hck、Lyn화SSeCKS재소서Kupffer세포중균유표체，용60%CCl4손상액자격후Hck、Lyn표체명현승고(P<0.01)，SSeCKS표체감소，단차이무통계학의의(P＞0.05)；Western Blot결과현시재Kupffer세포중Hck、SSeCKS표체교위명현，Lyn미견명현표체；IHC결과현시Hck、Lyn화SSeCKS재대서정상간조직Kupffer중균유표체；재염증조직중Hck、Lyn적표체명현증가，이SSeCKS적표체각현저감소。재염증자격후Hck、Lyn적표체여Kupffer세포수정정상관，이SSeCKS적표체여Kupffer세포수정부상관。결론：Hck、Lyn화SSeCKS삼여료NASH적발생발전，가작위NASH염증반응적잠재조공파점；검측기표체유조우NASH염증정도적판단，위림상상치료NASH제공일정적객관의거。
Objective:To analyze the significance and expression of Src family related kinases (Hck, Lyn)/SSeCKS in Kupffer cells. Methods: Real-time PCR was used to detect the expression of Hck,Lyn and SSeCKS in both normal and inflammation Kupffer cells. The expression of Hck, Lyn and SSeCKS were detected by Western Blot. The normal SD rats were treated with high-fat for 8 weeks and the models were established. The expression of Hck、Lyn and SSeCKS in kupffers cells were expressed by immunohistochemistry(IHC). Results:RT-PCR showed that Hck, Lyn and SSeCKS were all expressed in Kupffer cells in mice. The expression of Hck, Lyn increased significantly (P<0.05) after stimulating with 60% CCl4 liquid damage, the expression of SSeCKS were decreased, but without statistical significance(P>0.05);Western Blot showed that Hck, Lyn and SSeCKS were all expressed in Kupffer cells, especially the expressoin of Hck and SSeCKS was obvious and it has no obvious expression of Lyn in rat Kupffer cells. IHC showed that Hck, Lyn and SSeCKS were all expressed in normal liver tissue Kupffer cells in rats. In the expression of inflammatory tissue Hck and Lyn increased significantly, but the SSeCKS expression is significantly reduced. The expression of Hck and Lyn were positively correlated with number of Kupffer cells, while the expression of SSeCKS was negatively correlated with number of Kupffer cells after the inflammation stimulation. Conclusion:Hck, Lyn and SSeCKS are involved in the occurrence and development of NASH, that can be used as a potential target of NASH inflammation. The study can help clinical diagnosis of NASH inflammation degree, provide some objective basis for clinical treatment of NASH.
