A Crude 1-DNJ Extract from Home Made Bombyx Batryticatus Inhibits Diabetic Cardiomyopathy-Associated Fibrosis in db/db Mice and Reduces Protein N-Glycosylation Levels.
A Crude 1-DNJ Extract from Home Made Bombyx Batryticatus Inhibits Diabetic Cardiomyopathy-Associated Fibrosis in db/db Mice and Reduces Protein N-Glycosylation Levels.
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自制家蚕粗制 1-DNJ 提取物可抑制 db/db 小鼠糖尿病心肌病相关纤维化并降低蛋白质 N-糖基化水平
DOI：
10.3390/ijms19061699
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发表时间：
2018-06-07
影响因子：
5.6
通讯作者：
Ying WT
中科院分区：
文献类型：
--
作者：
Zhao Q;Jia TZ;Cao QC;Tian F;Ying WT
中药僵蚕，又名白色僵蚕，在中国临床上已被广泛使用数千年。它以其解痉和促进血液循环的作用而闻名。心肌细胞肥大、间质细胞增生和心肌纤维化与关键蛋白的N-糖基化密切相关。目的观察糖尿病早期心肌N-糖基化的变化，并探讨从BB中提取的1-脱氧野尻霉素（1-DNJ）的治疗作用。我们使用db/db（糖尿病）小鼠模型和基于亲水性色谱固相萃取与液相色谱质谱仪（LC-MS）鉴定策略集成的方法，以进行位点特异性N-甲基-N-（2-氨基乙基）左心室心肌细胞蛋白的糖基化分析。采用酶联免疫吸附法（ELISA）检测晚期糖基化终产物（AGEs）、羟脯氨酸、结缔组织生长因子（CTGF）等血清生化指标。此外，还采用透镜culinaris凝集素（LCA）凝集素印迹和异硫氰酸荧光素（FITC）标记凝集素亲和组织化学分析了N-聚糖的α-1，6-岩藻糖基化。结果表明，1-DNJ明显下调db/db小鼠心肌蛋白N-糖基化水平。1-DNJ可显著降低血清指标和纤维化相关细胞因子的表达水平，并呈剂量依赖性。db/db小鼠心肌糖链α-1，6-岩藻糖基化水平升高，1-DNJ对N-糖链α-1，6-岩藻糖基化的干预作用显著。为了验证这一结果，选择了众所周知的转化生长因子-β（TGF-β）/Smad 2/3途径，并采用蛋白质印迹法半定量分析核心α-1，6-岩藻糖基化TGF-β受体II（TGFR-βII）。结果支持LCA凝集素亲和组织化学和凝集素印迹分析的结论。结果表明，1-DNJ对FUT 8的表达无明显抑制作用。因此，1-DNJ减轻糖尿病性心肌病（DCM）相关纤维化的机制可以归结为抑制N-乙酰葡萄糖胺（N-GlcNAc）的形成和降低底物浓度。
The traditional Chinese drug Bombyx Batryticatus (BB), which is also named the white stiff silkworm, has been widely used in Chinese clinics for thousands of years. It is famous for its antispasmodic and blood circulation-promoting effects. Cardiomyocyte hypertrophy, interstitial cell hyperplasia, and myocardial fibrosis are closely related to the N-glycosylation of key proteins. To examine the alterations of N-glycosylation that occur in diabetic myocardium during the early stage of the disease, and to clarify the therapeutic effect of 1-Deoxynojirimycin (1-DNJ) extracted from BB, we used the db/db (diabetic) mouse model and an approach based on hydrophilic chromatography solid-phase extraction integrated with an liquid Chromatograph Mass Spectrometer (LC-MS) identification strategy to perform a site-specific N-glycosylation analysis of left ventricular cardiomyocyte proteins. Advanced glycation end products (AGEs), hydroxyproline, connective tissue growth factor (CTGF), and other serum biochemical indicators were measured with enzyme-linked immunosorbent assays (ELISA). In addition, the α-1,6-fucosylation of N-glycans was profiled with lens culinaris agglutinin (LCA) lectin blots and fluorescein isothiocyanate (FITC)-labelled lectin affinity histochemistry. The results indicated that 1-DNJ administration obviously downregulated myocardium protein N-glycosylation in db/db mice. The expression levels of serum indicators and fibrosis-related cytokines were reduced significantly by 1-DNJ in a dose-dependent manner. The glycan α-1,6-fucosylation level of the db/db mouse myocardium was elevated, and the intervention effect of 1-DNJ administration on N-glycan α-1,6-fucosylation was significant. To verify this result, the well-known transforming growth factor-β (TGF-β)/Smad2/3 pathway was selected, and core α-1,6-fucosylated TGF-β receptor II (TGFR-βII) was analysed semi-quantitatively with western blotting. The result supported the conclusions obtained from LCA lectin affinity histochemistry and lectin blot analysis. The expression level of α-1,6-fucosyltransferase (FUT8) mRNA was also detected, and the results showed that 1-DNJ administration did not cause obvious inhibitory effects on FUT8 expression. Therefore, the mechanism of 1-DNJ for relieving diabetic cardiomyopathy (DCM)-associated fibrosis can be concluded as the inhibition of N-acetylglucosamine (N-GlcNAc) formation and the reduction of substrate concentration.
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影响因子：
8.2
作者：
Bugger, Heiko;Abel, E. Dale
通讯作者：
Abel, E. Dale
影响因子：
6.1
作者：
Kang, Young Sun;Lee, Mi Hwa;Cha, Dae Ryong
通讯作者：
Cha, Dae Ryong
影响因子：
5
作者：
Higai, K;Azuma, Y;Matsumoto, K
通讯作者：
Matsumoto, K
影响因子：
2.8
作者：
AlJaroudi, Wael A.;Refaat, Marwan M.;Habib, Robert H.;Al-Shaar, Laila;Singh, Madhurmeet;Gutmann, Rebecca;Bloom, Heather L.;Dudley, Samuel C.;Ellinor, Patrick T.;Saba, Samir F.;Shalaby, Alaa A.;Weiss, Raul;McNamara, Dennis M.;Halder, Indrani;London, Barry
通讯作者：
London, Barry
DOI：
10.1152/ajprenal.00266.2002
发表时间：
2003-04-01
影响因子：
4.2
作者：
Kitsiou, PV;Tzinia, AK;Tsilibary, EC
通讯作者：
Tsilibary, EC