Salt Sensitive Hypertension Intervention New Targets Of Exploratory Research

Objective:Hypertension is a major public health challenge due to its high prevalence and concomitant increase in the risk for cardiovascular disease (CVD) and all-cause mortality. As a complex trait, hypertension is influenced by multiple environmental and genetic determinants, as well as their interactions. Among environmental determinants, dietary sodium intake is the most common and important risk factor for hypertension. However, there is substantial evidence suggesting that BP responses to dietary sodium intake vary considerably among individuals, a phenomenon described as salt sensitivity of blood pressure (BP). Elucidation of the genetic contribution to salt sensitivity provides important information regarding how genes and dietary sodium interact to influence BP. Salt-sensitive hypertension is, at least in part, under genetic control but the underlying genetic mechanisms are not fully clarified. Epigenetic control mechanisms may additively or synergistically interact to precisely respond to internal and external environmental cues. LncRNAs, tentatively defined as noncoding RNAs more than200nt in length, are characterized by the complexity and diversity of their sequences and mechanisms of action. A handful of studies have implicated LncRNAs in a variety of disease states. However, there are only preliminary studies on the role of LncRNAs in regulating genes that have been associated with salt-sensitive hypertension. In fact, there were virtually no published data on the overall pathophysiological contributions of LncRNAs to salt-sensitive hypertension. In this study, we identified the effects of these three LncRNAs, HOTAH、NOS3AS、aHIF, in the development of salt-sensitive hypertension in vivo and in vitro.Methods:We chose borderline hypertensive rat (BHR) as the animal model of this study. It is a kind of hypertension model which was induced by environmental factors, which is the F1generations of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). Since the sensitivity for different environmental factors is determined by mother of offspring, we select the progenies of SHR rats who is more sensitive.5weeks old sBHR randomly divided into four groups (four males and four females):high salt diet, high salt diet and Lycium barbarum L., control, low salt diet. After4weeks of normal diet, pure component feed which has different salt concentration was given to the four groups.The systolic blood pressure was monitored by tail cuff method. The renal endothelial and epithelial cells of sBHR were separated by magnetic bead coated with specific antibody. The expression of the LncRNAs (HOTAIR、NOS3AS、aHIF) and the coding genes(LSD1、eNOS、HIF-la) which were regulated by the LncRNAs were determined by real-time qPCR. They also were identified by real-time qPCR from human umbilical vein endothelial cells (HUVEC) and renal tubular cells (HKC) which were under high salt load In vitro.Results:We found that the blood pressure of high salt diet group and Lycium barbarum L. group was higher than that of control after having high salt diet for12weeks. Then the blood of Lycium barbarum L. group returned to normal after intervention of Lycium barbarum L. And the high salt diet exerts significant influence on the upregulation of LncRNA NOS3AS which was association with the increased blood level. The expression of eNOS mRNA was decreasing when the LncRNA NOS3AS was downregulation in the renal endothelial cells from Lycium barbarum L. group. The expression of LncRNA NOS3AS from HUVEC cells was increasing upon high salt treatment150mM NaC1). And the upregulation of LncRNA NOS3AS was earlier than the downregulation of eNOS mRNA. Importantly, Lycium barbarum polysaccharides (LBP) functionally regulated LncRNA NOS3A expression during high salt treatment(150mM NaCl), since LBP blunted it’s rising during prolonged high salt treatment. But the other LncRNA (HOTAIR and aHIF) had no obvious correlation with salt sensitive hypertension.Conclusions:The upregulation of LncRNA NOS3AS was association with develpoment of salt sensitive hypertension by regulating eNOS mRNA post-transcriptionally during high salt load in vivo and in vitro. Objective:As a traditional Chinese herbal formula composed of19herbs, Zicao-Huafu-Tang (ZHT) has been external used as a high efficient anti-inflammatory and analgesic medicine to treat inflammatory diseases such as skin trauma, diabetic ulcers, toothache pain, haemorrhoids, as well as cancers, indicating its general immunomodulatory effects. The mechanism of its wide pharmacological effects has not been explored. Toll-like receptors signaling and MAPKs activation have been known to modulate the inflammatory process in macrophage. We hypothesized that the favorable effects of ZHT on inflammation maybe through modulating this signaling pathway.Methods:We investigated the immunomodulatory mechanism of ZHT in Human monocyte cell line (THP-1) which was treated with ZHT in vitro, since macrophages are essential for innate immunity and play a central role in inflammation. Our hypothesis was tested in THP-1which was treated with ZHT at dosage1%,2%and3%(v/v)respectively, for24hours,48hours, and72hours. Cytokines, including tumour necrosis factor-a (TNF-a), Transforming growth factor-β1(TGF-β1), interleukin (IL)-6、IL-8and chemokine (C-C motif) ligand2(CCL2) in the cell culture supernatant, were determined by ELISA. The expression profile of M2b/c regulatory macrophages specific cytokines (IL-12low and IL-10high) and Toll-like receptors, the key players in immunomodulatory signaling pathway, were determined by real-time qPCR.Results:We found that ZHT exerts significant influence on TLR2/TLR4mediated polarization of macrophage subsets toward M2b/c regulatory macrophage phenotype. This may explain, at least in part, the favorable effects of ZHT on immune response, supported by evidence that more interleukin (IL)-10, IL-8, chemokine (C-C motif) ligand2(CCL2), Transforming growth factor-β2(TGF-β2), but less IL-12, IL-6, tumour necrosis factor-α (TNF-α), TGF-β1were released upon ZHT treatment. The underlying intracellular regulatory signaling mechanisms are complex since different cytokine and chemokine are regulated by different signaling intermediate activation. Our results support that MAPK activity is necessary for expression of IL-8and TGF-β2, while PKA activity is also necessary for expression of IL-10and TNF-α from other study reports. The main task of M2b/c regulatory macrophages is to dampen and control immune responses through an IL-12low and IL-10high expression profile and thereby contributing to the resolution of inflammatory responses. The M2b/c regulatory macrophages subpopulations are generated by different stimuli which need two stimuli to induce their anti-inflammatory activity. The first signal (for example, immune complexes, prostaglandins, adenosine or apoptotic cells) generally has little or no stimulatory function on its own. However, when combined with a second stimulus, such as a TLR ligands, the two signals reprogramme macrophages to produce IL-10, the production of which is the most important and reliable characteristic of regulatory macrophages. This is in good agreement with our observations here, that ZHT enhances IL-10expression but limits IL-12production. Of greater interest in the present study is that ZHT can induce M2b/c regulatory macrophages polarized states directly rather than reprogramming with the two signals reprogramme.Conclusions:ZHT not only suppressed inflammatory process, but also activated regulatory macrophages and facilitated inflammation resolution through modulating TLR2and TLR4signaling with MAPKs activation. These findings suggest that ZHT may offer some clinical advantages over glucocorticoids because it is likely to trigger activation of M2b/c regulatory macrophages that could be deemed proresolution but less levels of type1cytokines like TNF-a and IL-12. Objective:Contrary to common knowledge of the deleterious effects of obesity, it has been reported that obese stroke survivors are likely to have lower mortality than their underweight counterparts. Such a paradoxical phenomenon of lower mortality or risk of recurrent vascular disease in overweight or obese patients with established disease was coined the obesity paradox. However, in spite of accumulating reports on populations with various diseases, there are still doubts about the obesity paradox. Since the criteria for defining obesity and metabolic syndrome need to consider the influence of ethnicity, it is necessary to demonstrate the obesity paradox of stroke prognosis in Chinese population. In addition, the use of both body mass index (BMI) as an index of obesity has been challenged from recent studies as being not truely representative of total fat distribution. In this context, we evaluated the association between obesity and survival in patients with first-ever stroke by using BMI and waist to hip ratio (WHR) as indicators of obesity.Methods:We reviewed data from the multicenter study for assessment of risk factors of stroke sponsored by the Ministry of Science and Technology of China (973project). Initially,2000cases were recruited. Before data assessment,177subjects were excluded at different experimental stages because of lack of a definite diagnosis (24cases), absence of plasma (76cases), and failure of genotyping (77cases). Among the1823cases,807were diagnosed as thrombosis;513as lacunar; and503as hemorrhage. Participants were grouped into quartiles of BMI (Q1to Q4). The standards of BMI in adults in China was18.5≤BMI<24kg/m2for normal,24.0≤BMI<28kg/m2for overweight and BMI>28.0for obesity, then the participants were further classified as low weight, normal weight, overweight and obesity respectively. And the Chinese-specific of WHR cutoff is0.90for men and0.85for women, then the participants were divided into2groups according to WHR:normal weight (WHR<0.90for man, WHR<0.85for women), and obese (WHR>0.90for man, WHR≥0.85for women). Overall survival during follow-up was the primary end point. Mortality was assessed by Cox proportional hazard analysis and hazard ratios (HR) and95%confidence intervals (CI) are presented.Results:According to BMI (Q1to Q4), compared to controls in the lowest quartile, all-cause mortality risk was lower in the BMI Q3[24.2≤BMI<26.4kg/m2, hazard ratio (HR):0.77,95%confidence interval (CI):0.66-0.90),p=0.001]. Of1823eligible subjects,231(13%) were obese. After adjusting for confounders, compared with the normal weight patients, the lower all-cause mortality risk was in overweight patients (HR:0.75;95%CI:0.58-0.96, p=0.021), but low weight patients had higher all-cause mortality risk(HR:1.70;95%CI:1.09-2.65, p=0.02). And being overweight patients of thrombosis was associated with lower all-cause mortality risk (HR:0.65;95%CI:0.45-0.93,p=0.020) and lower risk of death caused by major vascular event (HR:0.56;95%CI:0.32-0.98, p=0.042). being low weight patients of lacunar was associated with the higher all-cause mortality risk (HR:6.11;95%CI:1.65-22.60,p=0.007) and higher risk of death caused by major vascular event (HR:9.91;95%CI:1.59-61.62, p=0.014). Based on WHR, after adjusting for confounders, compared with the normal group, being central obesity (HR:0.73;95%CI:0.58-0.91,p=0.006) was not only associated with decreased all-cause mortality risk, but also associated with lower risk of death caused by major vascular event(HR:0.65;95%CI:0.48-0.89,p=0.008). And being central obesity patients of lacunar (HR:0.45;95%CI:0.22-0.94,p=0.032) and hemorrhage (HR:0.59;95%CI:0.36-0.98,p=0.039) was associated with lower risk of death caused by major vascular event.Conclusions:based on the standards of BMI and WHR in China, obese and overweight stroke patients have significantly better survival rates compared to their leaner counterparts, supporting the widely held notion of the existence of a cardiovascular "obesity paradox."...