Sodium Pump Inhibitors In Blood Pressure Regulation

我们研究了以下三组(n=6)：低盐(0.3%氯化钠)饮食(LS)的DS；高盐(8%NaCl)饮食7周的DS(HS)；高盐饮食7周的DS，然后用单一的3E9抗MBG单抗治疗5天(HSAB)。检测左心室(LV)和主动脉中MBG水平、促肝纤维化信号转导相关蛋白水平和mRNA表达(基因表达配对分析、基因芯片分析、QIAGEN)。 HS组较LS组血压升高74 mm Hg(p&lt；0.01)，血浆MBG增加一倍(p&lt；0.05)，肾脏MBG排泄量增加6倍(p&lt；0.01)，组织重量增加(LV：2.37+/-0.05 vs.1.62+/-0.04 g/kg bw，p&lt；0.01；主动脉：4.44+/-0.17 vs.3.01+/-0.06 mg/mm*kg bw，p&lt；0.01；0.01)，LV胶原蛋白上涨3.5倍。与HS组相比，一次腹腔注射抗高血压DS单抗(HSAB组)5天后血压降低35 mm Hg(p&lt；0.01)，并显著降低左心室和主动脉重量(p&lt；0.01)。在高血压DS中，有一种组织特异性的基因表达上调模式，涉及转化生长因子信号转导(LV：TGFb1、TGFb2、MAPK3、CTGF、Smads、胶原-1(表1)；主动脉：TGFb2、TGFB3、PDGF、纤维连接蛋白、SNAIL1、PCOLCE、胶原蛋白；表2)，经MBG免疫中和后下调。这些观察结果得到了蛋白质印迹和qPCR分析的支持。 因此，MBG在DS中的免疫中和作用与下调盐敏感型高血压中MBG启动的转化生长因子诱导的纤维化相关的基因相关，从而产生抗重构作用。 ================================================================= 表1.DS患者左室基因表达的配对分析(Z比率)： 基因：_Hs/LS与HSAB/Hs：_基因描述： MAPK3_1.64与-2.66_丝裂原活化蛋白激酶3 TGFb1_1.72 vs.-2.20_转化生长因子β1 TGFb2_2.57 vs.-4.27_转化生长因子β2 CTGF_2.04与-4.90_结缔组织生长因子 Col1a2_0.80 vs.-2.35_胶原蛋白，类型1，阿尔法2 ================================================================= 表2.DS患者主动脉中基因表达的配对分析(Z比率)： 基因：_Hs/LS与HSAB/Hs：_基因描述 TGFb2_2.46与-1.60_转化生长因子β2 TGFB3_3.36 vs.-2.13_转化生长因子β3 PDGFa_1.08与-1.00_血小板衍生生长因子，α-多肽 PDGFRA_2.58与-2.43_血小板衍生生长因子受体，α多肽 FN_4.75 vs.-4.14_纤维连接蛋白 ITG1b_2.14与-2.16_纤维连接蛋白受体β1 Snai1_2.29 vs.-4.98_Snail 1同源基因 PCOLCE_2.55与-2.74_前胶原C-内肽酶增强子 Col5a2_2.95 vs.-3.08_胶原蛋白，类型5，阿尔法2 COL11A1_1.75 vs.-5.74_胶原蛋白，类型11，阿尔法1 Col16a1_2.11 vs.-2.44_胶原蛋白，类型16，阿尔法1 COL17A1_1.76 vs.-1.10_胶原蛋白，17型，阿尔法1 Col18a1_2.52 vs.-1.80_胶原蛋白，18型，阿尔法1

We studied the following three groups (n=6 each): DS on a low (0.3% NaCl) salt diet (LS); DS on a high salt (8% NaCl) diet for 7 weeks (HS); and DS on a high salt diet for 7 weeks, followed by a single 3E9 monoclonal anti-MBG antibody treatment for 5 days (HSAB). Levels of MBG and levels of proteins implicated in pro-fibrotic signaling, and mRNA expression (paired analysis of gene expression, microarray analysis, Qiagen) in left ventricles (LV) and aortae were assessed. In HS vs. LS, BP increased by 74 mmHg (p < 0.01), plasma MBG doubled (p < 0.05), renal MBG excretion increased 6-fold (p < 0.01), tissue weights increased (LV: 2.37 +/- 0.05 vs. 1.62 +/- 0.04 g/kg BW, p < 0.01; aorta: 4.44 +/- 0.17 vs. 3.01 +/- 0.06 mg/mm*kg BW, p < 0.01), and LV collagen rose 3.5-fold. A single intra-peritoneal administration of anti-MBG monoclonal antibody to hypertensive DS (HSAB group) reduces BP by 35 mmHg (p < 0.01) after 5 days, and markedly reduces collagen-1 and weights of left ventricles and aortae (p<0.01) vs. HS group. In hypertensive DS there was a tissue-specific pattern of up-regulation of expression of genes, implicated in TGF-signaling (LV: TGFb1, TGFb2, MAPK3, CTGF, SMADs, collagen-1 (Table 1); aorta: TGFb2, TGFb3, PDGF, fibronectin, SNAIL1, PCOLCE, collagens; Table 2), that was down-regulated following immunoneutralization of MBG. These observations were supported by western blot and qPCR analyses. Thus, immunoneutralization of MBG in DS produces an anti-remodeling effect associated with down-regulation of genes implicated in TGF-induced fibrosis initiated by MBG in salt-sensitive hypertension. ================================================================= Table 1. Paired analysis of gene expression (Z-ratio) in LV of DS: Gene:___HS/LS vs. HSAB/HS:___Gene description: MAPK3___1.64 vs. -2.66___Mitogen activated protein kinase 3 TGFb1___1.72 vs. -2.20___Transforming growth factor beta 1 TGFb2___2.57 vs. -4.27___Transforming growth factor beta 2 CTGF___2.04 vs. -4.90___Connective tissue growth factor Col1a2___0.80 vs. -2.35___Collagen, type 1, alpha 2 ================================================================= Table 2. Paired analysis of gene expression (Z-ratio) in aorta of DS: Gene:___HS/LS vs. HSAB/HS:___Gene description TGFb2___2.46 vs. -1.60___Transforming growth factor beta 2 TGFb3___3.36 vs. -2.13___Transforming growth factor beta 3 PDGFa___1.08 vs. -1.00___Platelet-derived growth factor, alpha polypeptide PDGFra___2.58 vs. -2.43___Platelet-derived growth factor receptor, alpha polypeptide FN______4.75 vs. -4.14___Fibronectin ITG1b____2.14 vs. -2.16___Fibronectin receptor beta 1 SNAI1____2.29 vs. -4.98___Snail 1 homolog PCOLCE___2.55 vs. -2.74___Procollagen C-endopeptidase enhancer Col5a2___2.95 vs. -3.08___Collagen, type 5, alpha 2 Col11a1___1.75 vs. -5.74___Collagen, type 11, alpha 1 Col16a1___2.11 vs. -2.44___Collagen, type 16, alpha 1 Col17a1___1.76 vs. -1.10___Collagen, type 17, alpha 1 Col18a1___2.52 vs. -1.80___Collagen, type 18, alpha 1