Huangqi Decoction,a compound Chinese herbal medicine,inhibits the proliferation and activation of hepatic stellate cells by regulating the long noncoding RNA-C18orf26-1/microRNA-663a/transforming growth factor-βaxis

Objective Huangqi Decoction(HQD),a classical traditional Chinese medicine formula,has been used as a valid treatment for alleviating liver fibrosis;however,the underlying molecular mechanism is still unknown.Although our previous studies showed that microRNA-663a(miR-663a)suppresses the proliferation and activation of hepatic stellate cells(HSCs)and the transforming growth factor-β/small mothers against decapentaplegic(TGF-β/Smad)pathway,whether long noncoding RNAs(lncRNAs)are involved in HSC activation via the miR-663a/TGF-β/Smad signaling pathway has not yet reported.The present study aimed to investigate the roles of lncRNA lnc-C18orf26-1 in the activation of HSCs and the mechanism by which HQD inhibits hepatic fibrosis.Methods The expression levels of lnc-C18orf26-1,miR-663a and related genes were measured by quantitative reverse transcription-polymerase chain reaction.HSCs were transfected with the miR-663a mimic or inhibitor and lnc-C18orf26-1 small interfering RNAs.The water-soluble tetrazolium salt-1 assay was used to assess the proliferation rate of HSCs.Changes in lncRNA expression were evaluated in miR-663a-overexpressing HSCs by using microarray to identify miR-663a-regulated lncRNAs.RNA hybrid was used to predict the potential miR-663a binding sites on lncRNAs.Luciferase reporter assays further confirmed the interaction between miR-663a and the lncRNA.The expression levels of collagen α-2(I)chain(COL1A2),α-smooth muscle actin(α-SMA)and TGF-β/Smad signaling pathway-related proteins were determined using Western blotting.Results Lnc-C18orf26-1 was upregulated in TGF-β1-activated HSCs and competitively bound to miR-663a.Knockdown of lnc-C18orf26-1 inhibited HSC proliferation and activation,downregulated TGF-β1-stimulatedα-SMA and COL1A2 expression,and inhibited the TGF-β1/Smad signaling pathway.HQD suppressed the proliferation and activation of HSCs.HQD increased miR-663a expression and decreased lnc-C18orf26-1 expression in HSCs.Further studies showed that HQD inhibited the expression of COL1A2,α-SMA,TGF-β1,TGF-βtype I receptor(TGF-βRI)and phosphorylated Smad2(p-Smad2)in HSCs,and these effects were reversed by miR-663a inhibitor treatment.Conclusion Our study identified lnc-C18orf26-1 and miR-663a as promising therapeutic targets for hepatic fibrosis.HQD inhibits HSC proliferation and activation at least partially by regulating the lnc-C18orf26-1/miR-663a/TGF-β1/TGF-βRI/p-Smad2 axis.

血清细胞角蛋白19片段21-1、E26转录因子-1联合^(18)F-FDG PET/CT对胃癌术后复发的诊断价值

目的探讨血清细胞角蛋白19片段抗原21-1(CYFRA21-1)、E26转录因子-1(Ets-1)检测联合^(18)F-氟代脱氧葡萄糖(^(18)F-FDG)正电子发射断层显像/CT(PET/CT)对胃癌术后复发的诊断价值。方法选取2018年1月至2020年1月在上海中医药大学附属曙光医院普外科行手术切除治疗的178例胃癌患者为研究对象,术后随访截止时间2021年2月,无死亡病例。根据再次手术后病理检查结合临床随访结果分为术后复发组(31例)和术后未复发组(147例)。分别采用电化学发光法、化学发光酶联免疫分析法检测所有受试者血清CYFRA21-1、Ets-1水平,并进行^(18)F-FDG PET/CT检查,测定半定量参数原发灶最大标准摄取值(SUV_(max)),并绘制受试者工作特征曲线(ROC曲线)评估血清CYFRA21-1、Ets-1水平、SUV_(max)指标单独及三者联合对胃癌术后复发的诊断价值。结果术后复发组血清CYFRA21-1、Ets-1水平、SUV_(max)值均高于术后未复发组[(5.79±0.61)μg/L比(2.68±0.29)μg/L、(6.16±0.63)μg/L比(2.78±0.31)μg/L、(3.81±0.41)比(1.38±0.15)](P<0.01)。术后复发患者血清CYFRA21-1、Ets-1水平、SUV_(max)值最佳截断点分别为3.45μg/L、3.51μg/L、2.56,三项指标单独检测诊断胃癌术后复发的ROC曲线下面积分别为0.713(95%CI 0.674~0.742)、0.709(95%CI 0.617~0.737)、0.715(95%CI 0.676~0.749),灵敏度分别为77.42%、74.19%、80.65%,特异度分别为68.71%、66.67%、69.39%,准确度分别为70.22%、67.98%、71.35%;三项指标联合诊断胃癌术后复发的ROC曲线下面积为0.778(95%CI 0.724~0.846),灵敏度为67.74%、特异度为85.03%、准确度为82.02%,三项联合检测诊断胃癌术后复发的特异度、准确度高于单独诊断(P<0.05)。结论血清CYFRA21-1、Ets-1水平及SUV_(max)值联合诊断胃癌术后复发较单一指标诊断效能更高。