纤调蛋白对氧化应激诱导的胰腺星形细胞活化的影响

Effects of oxidative stress on the activation of pancreatic stellate cells through the up-regulation of fibromodulin

目的探讨纤调蛋白（FMOD）在氧化应激诱导的胰腺星形细胞（PSCs）活化中的作用。方法应用靶向FMOD-shRNA（sh-FMOD）慢病毒感染PSCs，再用促氧化剂甲萘醌（MND）处理48 h（MND＋sh-FMOD组），以等容积DMSO处理的慢病毒感染的PSCs细胞为sh-FMOD组，亲本细胞为对照组，MND处理的PSCs为MND组。采用RT-PCR法检测PSCs活化标志物α-平滑肌肌动蛋白（α-SMA）、Ⅲ型胶原蛋白α1（Col3α1）、Ⅰ型胶原蛋白α1（Col1α1）、金属蛋白酶抑制剂1（TIMP1）、整合素α1（α1-Integrin）mRNA表达。采用二丁基二氯化锡尾静脉注射建立慢性胰腺炎（CP）大鼠模型。应用免疫组织化学染色法检测大鼠正常胰腺组织和CP胰腺组织中FMOD、α-SMA及抗氧化标志物超氧化物歧化酶（SOD）、氧化标志物丙二醛（MDA）蛋白表达。结果sh-FMOD组PSCs的FMOD mRNA表达量较对照组显著下降（0.16±0.03比1），差异有统计学意义（P〈0.01），表明FMOD基因沉默成功。MND组PSCs的FMOD、α-SMA、Col3α1、Col1α1、TIMP1、α1-Integrin mRNA表达量均较对照组显著增加，而MND＋sh-FMOD组PSCs的上述各基因表达量均较MND组下降，差异均有统计学意义（P值〈0.05或〈0.01）。CP胰腺组织FMOD、α-SMA、SOD、MDA蛋白表达均较正常胰腺组织增加，差异具有统计学意义（P值均〈0.05）。结论氧化应激通过诱导FMOD表达促进PSCs活化。

ObjectiveTo investigate the role of fibromodulin （FMOD） in the xidative stress induced activation of pancreatic stellate cells （PSCs）. MethodsLentivirus containing ShRNA targeting FMOD （sh-FMOD） was transfected into PSCs, and then the prooxidant menadione （MND） was used to treat PSCs for 24 h （MND＋ sh-FMOD group）. Lentivirus transfected PSCs cells treated by a equal volume of DMSO served as sh-FMOD group, parent cells as control group and PSCs treated by MND as MND group. RT-PCR were used to detect the mRNA expression of the markers of activated PSCs including α-SMA, Col3α1, Col1α1, TIMP1 and α1-integrin. Chronic pancreatitis （CP） rat model was induced by DBTC infusion into the tail vein. Immunohistochemical （IHC） staining was used to detect the protein expression of FMOD, FN, α-SMA, SOD and MDA in normal pancreatic tissue and CP tissue. ResultsFMOD mRNA expression of the PSCs in FMOD group was obviously lowerer than that in control group （0.16±0.03 vs 1）, and the difference was statistically significant （P〈0.01）, indicating that FMOD was successfully silenced. The mRNA expression of FMOD, α-SMA, Col3α1, Col1α1, TIMP1 and α1-Integrin of PSCs in MND group was obviously higher than those in control group, which in MND＋ sh-FMOD group was lower than those in MND group, and the difference was statistically significant （P〈0.05 or 〈0.01）. Compared with those in normal pancreatic tissue, the protein expression of FMOD, α-SMA, SOD and MDA in CP tissue was up-regulated, and the difference was statistically significant （all P〈0.05）.ConclusionsOxidative stress can facilitate the activation of PSCs through the induction of fibromodulin expression.