目的探讨大黄-丹参药对(RS)调控肌腱蛋白C(TNC)/表皮生长因子受体(EGFR)/信号转导和转录活化因子3(STAT3)通路延缓单侧输尿管梗阻(UUO)大鼠肾纤维化的机制。方法50只雄性SD大鼠随机分为假手术组、模型组、尿毒清颗粒(阳性药,2.5 g·...目的探讨大黄-丹参药对(RS)调控肌腱蛋白C(TNC)/表皮生长因子受体(EGFR)/信号转导和转录活化因子3(STAT3)通路延缓单侧输尿管梗阻(UUO)大鼠肾纤维化的机制。方法50只雄性SD大鼠随机分为假手术组、模型组、尿毒清颗粒(阳性药,2.5 g·kg^(-1))组和RS低、高剂量(生药3.0、6.0 g·kg^(-1))组,每组10只。除假手术组外,其余各组大鼠采用UUO法制备肾间质纤维化模型。各给药组ig相应药物,假手术组、模型组ig等体积纯净水,连续14 d。代谢笼收集各组大鼠24 h尿液,腹主动脉取血分离血清,测定肌酐(Scr)和尿素氮(BUN)水平;酶联免疫吸附法(ELISA)检测尿液和血清中TNC的水平;HE和Masson染色观察各大鼠肾组织病理形态学变化;实时荧光定量法(qRT-PCR)检测肾组织TNC、EGFR、STAT3、TGF-β、纤维连接蛋白(FN)和α-平滑肌肌动蛋白(α-SMA)的m RNA表达;蛋白免疫印记法(Western blotting)检测肾组织TNC、EGFR、STAT3和磷酸化STAT3(p-STAT3)的蛋白表达;免疫组化染色(IHC)观察肾组织TNC、EGFR、p-STAT3的阳性表达。SD大鼠ig RS(生药6.0 g·kg^(-1))或纯净水制备含药或空白血清;以人肾皮质近曲小管上皮(HK-2)细胞为研究对象,将细胞分为对照组、模型组(TGF-β10 ng·mL-1)、RS含药血清(10%)(RS)组、过表达阴性对照(NC-OE)组、TNC过表达(TNC-OE)组、TNC过表达+RS含药血清(10%)(TNC-OE+RS)组。按相应条件培养后,qRT-PCR检测各组细胞TNC、EGFR和STAT3 mRNA的表达,Western blotting检测TNC、EGFR、STAT3和p-STAT3的表达。结果体内实验结果表明,与假手术组相比,模型组大鼠血清Scr、BUN、TNC水平和尿液TNC水平均明显升高(P<0.01);肾组织病理损伤加重,肾纤维化评分显著升高(P<0.01);肾组织TNC、EGFR、STAT3、TGF-β、FN和α-SMA m RNA水平和TNC、EGFR、p-STAT3蛋白表达水平及其阳性表达占比均明显上调(P<0.01)。与模型组相比,RS低、高剂量组和尿毒清颗粒组大鼠血清Scr、BUN、TNC水平和尿液TNC水平均显著降低(P<0.01);肾组织损伤减轻,纤维化评分明显降低(P<0.01);肾组织TNC、EGFR、STAT3、TGF-β、FN和α-SMA m RNA水平和TNC、EGFR、p-STAT3蛋白表达水平及其阳性表达占比均显著下调(P<0.05、0.01)。细胞实验结果表明,与对照组相比,模型组细胞TNC、EGFR、STAT3 m RNA表达及TNC、EGFR、p-STAT3蛋白表达水平明显升高(P<0.01);与模型组相比,RS组细胞TNC、EGFR、STAT3 m RNA表达及TNC、EGFR、p-STAT3蛋白表达水平显著降低(P<0.05、0.01)。与NC-OE组相比,TNC-OE组细胞TNC、EGFR、STAT3 m RNA表达及TNC、EGFR、p-STAT3蛋白表达水平明显升高(P<0.01);与TNC-OE组相比,TNC-OE+RS组细胞TNC、EGFR、STAT3 mRNA表达及TNC、EGFR、p-STAT3蛋白表达水平显著降低(P<0.01)。结论RS能够从体内、外延缓肾纤维化的进展,其作用机制可能与调控TNC/EGFR/STAT3通路有关。展开更多
Background: Pancreatic ductal adenocarcinoma(PDAC) is the most deadly type of tumor, and its pathogenesis remains unknown. Circular RNAs(circRNAs) may be functional and bind to micro RNAs and consequently, influence t...Background: Pancreatic ductal adenocarcinoma(PDAC) is the most deadly type of tumor, and its pathogenesis remains unknown. Circular RNAs(circRNAs) may be functional and bind to micro RNAs and consequently, influence the activity of targeted mRNAs. Recent researches indicate that one circRNA, ciRS-7, acts as a sponge of miR-7 and thus, inhibits its activity. It is well known that miR-7 is a cancer suppressor in many cancers. However, the relationship between ciRS-7 and miR-7, and the role of ciRS-7 in PDAC, remains to be elucidated. Methods: miR-7 and ciRS-7 expression in 41 pairs of PDAC tumors and their paracancerous tissues were detected by quantitative reverse transcription polymerase chain reaction(qRT-PCR). The relationships between their expression levels and clinicopathological features in PDAC tissues were assessed. The relationship between miR-7 and ciRS-7 was also assessed by Spearman’s correlation. We also used cell lines to evaluate the role of ciRS-7 in cell line behavior. The ciRS-7 interfere RNA(si RNA) and its empty vector were transfected into PDAC cells. PDAC cells proliferation and invasion abilities were detected by MTT assay and invasion analysis. The expression of proteins was assessed by Western blotting. Results: ciRS-7 expression was significantly higher in PDAC tissues than paracancerous tissues( P = 0.002). However, miR-7 expression showed the opposite trend( P = 0.048). Moreover, ciRS-7 expression was inversely correlated with miR-7 in PDAC( r s =-0.353, P = 0.023). ciRS-7 expression was also significantly elevated in venous invasion(3.72 ± 2.93 vs. 2.14 ± 1.26;P = 0.028) and lymph node metastasis(4.19 ± 2.75 vs. 2.32 ± 1.90;P = 0.016) in PDAC patients. Furthermore, ciRS-7 knockdown suppressed cell proliferation and invasion of PDAC cells( P < 0.05), and the downregulation of ciRS-7 resulted in miR-7 overexpression and subsequent inhibition of epidermal growth factor receptor(EGFR) and signal transducer and activator of transcription 3(STAT3). Conclusions: Circular RNA ciRS-7 plays an oncogene role in PDAC, partly by targeting miR-7 and regulating the EGFR/STAT3 signaling pathway.展开更多
溃疡性结肠炎(ulcerative colitis,UC)是一种以结肠黏膜及黏膜下层炎症为主要特征的慢性非特异性炎症,其致病机制复杂,易反复发作,现代医学研究认为其涉及氧化应激、免疫失衡等多方面因素。信号转导和转录激活因子3(signal transducer a...溃疡性结肠炎(ulcerative colitis,UC)是一种以结肠黏膜及黏膜下层炎症为主要特征的慢性非特异性炎症,其致病机制复杂,易反复发作,现代医学研究认为其涉及氧化应激、免疫失衡等多方面因素。信号转导和转录激活因子3(signal transducer and activator of transcription,STAT3)是调节细胞生长、分化和存活的重要因子,可被相关细胞因子激活,从而介导炎症、氧化应激及免疫反应以影响UC病理进程,并与核因子κB(nuclear factor kappa-B,NF-κB)、NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)、细胞因子信号传导抑制因子(suppressor of cytokine signaling,SOCSs)等信号通路存在串扰现象。STAT3作为近年来UC相关研究的热点之一,本文综述了中药通过调控STAT3信号通路防治UC的研究进展,深入探究了STAT3激活及介导UC病理过程的分子机制,以及中药成分如何通过多途径调控STAT3信号通路,发挥其潜在的作用机制。相关研究揭示了中药通过调节STAT3信号通路,不仅有效抑制炎症、氧化应激的发生,还能在调控免疫反应、维持肠道屏障功能及完整性等方面发挥重要作用,有望为治疗UC提供新思路。展开更多
文摘目的探讨大黄-丹参药对(RS)调控肌腱蛋白C(TNC)/表皮生长因子受体(EGFR)/信号转导和转录活化因子3(STAT3)通路延缓单侧输尿管梗阻(UUO)大鼠肾纤维化的机制。方法50只雄性SD大鼠随机分为假手术组、模型组、尿毒清颗粒(阳性药,2.5 g·kg^(-1))组和RS低、高剂量(生药3.0、6.0 g·kg^(-1))组,每组10只。除假手术组外,其余各组大鼠采用UUO法制备肾间质纤维化模型。各给药组ig相应药物,假手术组、模型组ig等体积纯净水,连续14 d。代谢笼收集各组大鼠24 h尿液,腹主动脉取血分离血清,测定肌酐(Scr)和尿素氮(BUN)水平;酶联免疫吸附法(ELISA)检测尿液和血清中TNC的水平;HE和Masson染色观察各大鼠肾组织病理形态学变化;实时荧光定量法(qRT-PCR)检测肾组织TNC、EGFR、STAT3、TGF-β、纤维连接蛋白(FN)和α-平滑肌肌动蛋白(α-SMA)的m RNA表达;蛋白免疫印记法(Western blotting)检测肾组织TNC、EGFR、STAT3和磷酸化STAT3(p-STAT3)的蛋白表达;免疫组化染色(IHC)观察肾组织TNC、EGFR、p-STAT3的阳性表达。SD大鼠ig RS(生药6.0 g·kg^(-1))或纯净水制备含药或空白血清;以人肾皮质近曲小管上皮(HK-2)细胞为研究对象,将细胞分为对照组、模型组(TGF-β10 ng·mL-1)、RS含药血清(10%)(RS)组、过表达阴性对照(NC-OE)组、TNC过表达(TNC-OE)组、TNC过表达+RS含药血清(10%)(TNC-OE+RS)组。按相应条件培养后,qRT-PCR检测各组细胞TNC、EGFR和STAT3 mRNA的表达,Western blotting检测TNC、EGFR、STAT3和p-STAT3的表达。结果体内实验结果表明,与假手术组相比,模型组大鼠血清Scr、BUN、TNC水平和尿液TNC水平均明显升高(P<0.01);肾组织病理损伤加重,肾纤维化评分显著升高(P<0.01);肾组织TNC、EGFR、STAT3、TGF-β、FN和α-SMA m RNA水平和TNC、EGFR、p-STAT3蛋白表达水平及其阳性表达占比均明显上调(P<0.01)。与模型组相比,RS低、高剂量组和尿毒清颗粒组大鼠血清Scr、BUN、TNC水平和尿液TNC水平均显著降低(P<0.01);肾组织损伤减轻,纤维化评分明显降低(P<0.01);肾组织TNC、EGFR、STAT3、TGF-β、FN和α-SMA m RNA水平和TNC、EGFR、p-STAT3蛋白表达水平及其阳性表达占比均显著下调(P<0.05、0.01)。细胞实验结果表明,与对照组相比,模型组细胞TNC、EGFR、STAT3 m RNA表达及TNC、EGFR、p-STAT3蛋白表达水平明显升高(P<0.01);与模型组相比,RS组细胞TNC、EGFR、STAT3 m RNA表达及TNC、EGFR、p-STAT3蛋白表达水平显著降低(P<0.05、0.01)。与NC-OE组相比,TNC-OE组细胞TNC、EGFR、STAT3 m RNA表达及TNC、EGFR、p-STAT3蛋白表达水平明显升高(P<0.01);与TNC-OE组相比,TNC-OE+RS组细胞TNC、EGFR、STAT3 mRNA表达及TNC、EGFR、p-STAT3蛋白表达水平显著降低(P<0.01)。结论RS能够从体内、外延缓肾纤维化的进展,其作用机制可能与调控TNC/EGFR/STAT3通路有关。
基金supported by grants from the Key Research and Development Program Projects in Anhui Province(1804h08020277)the Programs for Science and Technology Development of An-hui Province(1606c08234)
文摘Background: Pancreatic ductal adenocarcinoma(PDAC) is the most deadly type of tumor, and its pathogenesis remains unknown. Circular RNAs(circRNAs) may be functional and bind to micro RNAs and consequently, influence the activity of targeted mRNAs. Recent researches indicate that one circRNA, ciRS-7, acts as a sponge of miR-7 and thus, inhibits its activity. It is well known that miR-7 is a cancer suppressor in many cancers. However, the relationship between ciRS-7 and miR-7, and the role of ciRS-7 in PDAC, remains to be elucidated. Methods: miR-7 and ciRS-7 expression in 41 pairs of PDAC tumors and their paracancerous tissues were detected by quantitative reverse transcription polymerase chain reaction(qRT-PCR). The relationships between their expression levels and clinicopathological features in PDAC tissues were assessed. The relationship between miR-7 and ciRS-7 was also assessed by Spearman’s correlation. We also used cell lines to evaluate the role of ciRS-7 in cell line behavior. The ciRS-7 interfere RNA(si RNA) and its empty vector were transfected into PDAC cells. PDAC cells proliferation and invasion abilities were detected by MTT assay and invasion analysis. The expression of proteins was assessed by Western blotting. Results: ciRS-7 expression was significantly higher in PDAC tissues than paracancerous tissues( P = 0.002). However, miR-7 expression showed the opposite trend( P = 0.048). Moreover, ciRS-7 expression was inversely correlated with miR-7 in PDAC( r s =-0.353, P = 0.023). ciRS-7 expression was also significantly elevated in venous invasion(3.72 ± 2.93 vs. 2.14 ± 1.26;P = 0.028) and lymph node metastasis(4.19 ± 2.75 vs. 2.32 ± 1.90;P = 0.016) in PDAC patients. Furthermore, ciRS-7 knockdown suppressed cell proliferation and invasion of PDAC cells( P < 0.05), and the downregulation of ciRS-7 resulted in miR-7 overexpression and subsequent inhibition of epidermal growth factor receptor(EGFR) and signal transducer and activator of transcription 3(STAT3). Conclusions: Circular RNA ciRS-7 plays an oncogene role in PDAC, partly by targeting miR-7 and regulating the EGFR/STAT3 signaling pathway.
文摘溃疡性结肠炎(ulcerative colitis,UC)是一种以结肠黏膜及黏膜下层炎症为主要特征的慢性非特异性炎症,其致病机制复杂,易反复发作,现代医学研究认为其涉及氧化应激、免疫失衡等多方面因素。信号转导和转录激活因子3(signal transducer and activator of transcription,STAT3)是调节细胞生长、分化和存活的重要因子,可被相关细胞因子激活,从而介导炎症、氧化应激及免疫反应以影响UC病理进程,并与核因子κB(nuclear factor kappa-B,NF-κB)、NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)、细胞因子信号传导抑制因子(suppressor of cytokine signaling,SOCSs)等信号通路存在串扰现象。STAT3作为近年来UC相关研究的热点之一,本文综述了中药通过调控STAT3信号通路防治UC的研究进展,深入探究了STAT3激活及介导UC病理过程的分子机制,以及中药成分如何通过多途径调控STAT3信号通路,发挥其潜在的作用机制。相关研究揭示了中药通过调节STAT3信号通路,不仅有效抑制炎症、氧化应激的发生,还能在调控免疫反应、维持肠道屏障功能及完整性等方面发挥重要作用,有望为治疗UC提供新思路。
