Jingjing Cai

Regulator of G-Protein Signaling 10 Negatively Regulates Cardiac Remodeling by Blocking Mitogen-Activated Protein Kinase–Extracellular Signal-Regulated Protein Kinase 1/2 Signaling

Regulator of G-protein signaling 10 (RGS10) is an important member of the RGS family and produces biological effects in multiple organs. We used a genetic approach to study the role of RGS10 in the regulation of pathological cardiac hypertrophy and found that RGS10 can negatively influence pressure overload–induced cardiac remodeling. RGS10 expression was markedly decreased in failing human hearts and hypertrophic murine hearts. The extent of aortic banding–induced cardiac hypertrophy, dysfunction, and fibrosis in RGS10-knockout mice was exacerbated, whereas the heart of transgenic mice with cardiac-specific RGS10 overexpression exhibited an alleviated response to pressure overload. Consistently, RGS10 also inhibited an angiotensin II–induced hypertrophic response in isolated cardiomyocytes. Mechanistically, cardiac remodeling improvement elicited by RGS10 was associated with the abrogation of mitogen-activated protein kinase kinase 1/2–extracellular signal-regulated protein kinase 1/2 signaling. Furthermore, the inhibition of mitogen-activated protein kinase kinase–extracellular signal-regulated protein kinase 1/2 transduction abolished RGS10 deletion-induced hypertrophic aggravation. These findings place RGS10 and its downstream signaling mitogen-activated protein kinase kinase–extracellular signal-regulated protein kinase 1/2 as crucial regulators of pathological cardiac hypertrophy after pressure overload and identify this pathway as a potential therapeutic target to attenuate the pressure overload–driven cardiac remodeling.

Comparing Antihypertensive Effect and Plasma Ciclosporin Concentration between Amlodipine and Valsartan Regimens in Hypertensive Renal Transplant Patients Receiving Ciclosporin Therapy

BackgroundHypertension, a major complication in kidney transplant recipients, is associated with premature death and graft loss. However, an optimal antihypertensive therapy for these patients has not been established [Chinese Clinical Trial Registry No. ChiCTR-TRC-10001071].ObjectiveThe aim of the present study was to evaluate the effect of amlodipine and valsartan on BP control in renal transplant patients and to analyze the correlation between cytochrome P450 (CYP) 3A5 or multidrug resistance-1 gene (MDR1) genotype and the antihypertensive effect of these two regimens.Methods150 renal transplant patients with stage 1 or 2 hypertension were enrolled in the trial. Patients were randomly assigned to amlodipine or valsartan. Metoprolol was added if BP was not under control after 4 weeks. BP and plasma levels of ciclosporin were monitored during the 24-week trial. CYP3A5 and MDR1 genotypes were determined using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.ResultsThe demographic features and baseline BP were similar between these two groups. During the 24-week trial, the reduction of systolic BP (SBP) was similar between the amlodipine and valsartan groups. However, the reduction of diastolic BP (DBP) was significantly greater in the amlodipine group compared with the valsartan group at 12, 16, and 24 weeks of treatment. The plasma level of ciclosporin at 2 hours of medication was significantly higher in the amlodipine group than in the valsartan group after 4 weeks of the trial. The reduction of DBP at 24 weeks was greater in the subjects with CYP3A5 *3/*3 variant than in those with CYP3A5*1/*1 variant (−13.5± 1.9mmHg vs −8.7± 1.6mmHg, p<0.05).ConclusionThe present study demonstrated that amlodipine produced a greater reduction of DBP than valsartan, although both amlodipine and valsartan resulted in satisfactory control of BP in patients with renal transplantation. Administration of amlodipine significantly increased the plasma concentration of ciclosporin, and its effects on BP control and ciclosporin concentration may be associated with the CYP3A5 genotype in these subjects [Chinese Clinical Trial Registry No. ChiCTR-TRC-10001071].

Inhibition of PKR impairs angiogenesis through a VEGF pathway

Peripheral artery disease (PAD) is a common clinical problem, and its pathophysiological mechanisms are incompletely understood. Double-stranded RNA-activated protein kinase (PKR) is a ubiquitously expressed serine/threonine protein kinase. Although PKR has been reported in antivirus and the immune system, the role of PKR in vascular function, especially in angiogenesis, is still unclear. PKR(-/-) mice were used in our experiments. Blood flow recovery was significantly delayed in PKR(-/-) vs. WT mice (Laser Doppler detection, n = 9, P < 0.01), accompanied by 34% reduced CD31-positive stain in ischemic muscle 28 days after procedure (immunohistochemistry, n = 9, P < 0.05). PKR expression decreased in the first 12 h and increased to peak at 24 h in human umbilical vein endothelial cells (HUVECs) in response to hypoxia (Western blot analyses, n = 3, P < 0.05). Accordingly, phospho-PKR expression increased in HUVECs 24 h after treatment with hypoxia (Western blot analyses, n = 3, P < 0.05). Inhibition of PKR (siRNA transfection) reduced microtubule formation (Matrigel tube formation, n = 3, vs. control siRNA, P < 0.05) and migration (wound healing, n = 3, vs. control siRNA, P < 0.05) by 33 and 59%, respectively. Vascular endothelial growth factor (VEGF) expression in ischemic muscle from PKR(-/-) mice was significantly decreased by 54% 1 day after procedure (n = 3, P < 0.05, vs. WT) and by 63% 7 days after procedure (n = 3, P < 0.01, vs. WT), respectively. At the same time, VEGF expression in HUVECs decreased by 21% (n = 3, P < 0.05, PKR siRNA vs. control siRNA). These findings demonstrate that PKR mediates angiogenesis through a VEGF pathway, which may form the basis for future intervention of PAD.

Inferior myocardial infarction secondary to aortic dissection associated with bicuspid aortic valve

Aortic dissection (AD) is a life-threatening condition and may present with symptoms which mimic myocardial infarction, leading to misdiagnosis and inappropriate use of anticoagulant and thrombolytic therapy. A 40-year-old woman with no prior history presented in our emergency department with sudden chest pain. Electrocardiography (ECG) showed a ST-segment elevation in leads II, III and avF, suggesting an acute inferior myocardial infarction. The patient was given anticoagulation and antiplatelet treatment. Coronary angiography, transthoracic echocardiography and computed tomography were performed. The patient was diagnosed with DeBakey I aortic dissection extending from ascending aorta to iliac artery, and associated with bicuspid aortic valve.Surgical treatments with a replacement of the ascending aorta, aortic valve replacement and coronary artery bypass grafting were successfully performed. Early imaging examination, if possible, might assist the diagnosis and guide the management of this disease. The condition of myocardial infarction secondary to aortic dissection is discussed.

Hepatic leukocyte immunoglobulin‐like receptor B4 (LILRB4) attenuates nonalcoholic fatty liver disease via SHP1‐TRAF6 pathway

Nonalcoholic fatty liver disease (NAFLD) is an increasingly prevalent liver pathology characterized by hepatic steatosis and commonly accompanied by systematic inflammation and metabolic disorder. Despite an accumulating number of studies, no pharmacological strategy is available to treat this condition in the clinic. In this study, we applied extensive gain‐ and loss‐of‐function approaches to identify the key immune factor leukocyte immunoglobulin‐like receptor B4 (LILRB4) as a negative regulator of NAFLD. The hepatocyte‐specific knockout of LILRB4 (LILRB4‐HKO) exacerbated high‐fat diet–induced insulin resistance, glucose metabolic imbalance, hepatic lipid accumulation, and systematic inflammation in mice, whereas LILRB4 overexpression in hepatocytes showed a completely opposite phenotype relative to that of LILRB4‐HKO mice when compared with their corresponding controls. Further investigations of molecular mechanisms demonstrated that LILRB4 recruits SHP1 to inhibit TRAF6 ubiquitination and subsequent inactivation of nuclear factor kappa B and mitogen‐activated protein kinase cascades. From a therapeutic perspective, the overexpression of LILRB4 in a genetic model of NAFLD, ob/ob mice, largely reversed the inherent hepatic steatosis, inflammation, and metabolic disorder. Conclusion: Targeting hepatic LILRB4 to improve its expression or activation represents a promising strategy for the treatment of NAFLD as well as related liver and metabolic diseases. (Hepatology 2018;67:1303‐1319)

HIV protease inhibitors in pulmonary hypertension: rationale and design of a pilot trial in idiopathic pulmonary arterial hypertension

We propose an exploratory clinical study, the first of its kind to our knowledge, to determine the safety and potential clinical benefit of the combination of the HIV protease inhibitors (HIV-PIs) saquinavir and ritonavir (SQV+RIT) in patients with idiopathic pulmonary arterial hypertension (IPAH). This study is based on evidence that (1) HIV-PIs can improve pulmonary hemodynamics in experimental models; (2) both Toll-like receptor 4 and high-mobility group box 1 (HMGB1) participate in the pathogenesis of experimental pulmonary hypertension; and (3) a high-throughput screen for inhibitors of HMGB1-induced macrophage activation yielded HIV-PIs as potent inhibitors of HMGB1-induced cytokine production. In this proposed open-label, pre-post study, micro, low, and standard doses of SQV+RIT will be given to IPAH patients for 14 days. Patients will receive follow-up for the next 14 days. The primary outcome to be evaluated is change in HMGB1 level from baseline at 14 days. The secondary outcome is changes in tumor necrosis factor α, interleukin 1β, interleukin 6, C-reactive protein, pulmonary arterial pressure based on echocardiography parameters and New York Heart Association/World Health Organization functional class, and Brog dyspnea scale index from baseline at 14 days. Other secondary measurements will include N-terminal pro-brain natriuretic peptide, atrial natriuretic peptide, and 6-minute walk distance. We propose that SQV+RIT treatment will improve inflammatory disorders and pulmonary hemodynamics in IPAH patients. If the data support a potentially useful therapeutic effect and suggest that SQV+RIT is safe in IPAH patients, the study will warrant further investigation. (ClinicalTrials.gov identifier: NCT02023450.)

110 EFFECTS OF GENISTEIN ON REGULATION OF DIHYDROTESTOSTERONE-INDUCED CELL PROLIFERATION IN ENDOTHELIAL AND PROSTATE CANCER CELLS

Grant Supports: This study was supported in part by a grant from National Institutes of Health (NIH) (UL1 RR024996) and a grant from the National Natural Science Foundation of China (No. 30873126) and a grant from the National Basic Research Program of China (2011CB5120010. Background: Androgen deprivation therapy of prostate cancer with estrogens shows significant cardiovascular side effects. This study was to examine the effect of genistein comparing to 17&bgr; − estradiol on modulation of androgen actions in human aortic endothelial cells (HAECs) and prostate cancer LAPC-4 and LNCaP cells. Methods: MTS, RT-PCR, and Western blot were used to detect cell proliferation, mRNA and protein expression of estrogen receptor (ER) gene, and cyclin A genes, respectively. Specific ER&bgr; siRNA was synthesized and transfected to knockdown ER&bgr; expression in prostate cancer cells. Results: Dihydrotestosterone (DHT) produced a time and dose-dependent induction of cell proliferation in HAEC, LAPC-4 and LNCaP cells. These DHT actions were inhibited by genistein in LAPC-4 and LNCaP cells but not in HAEC cells. While &bgr;E2 only attenuated the DHT-induced cell proliferation in HAEC and LAPC-4 cells without any inhibition of DHT-induced LNCaP cell proliferation. In LAPC-4 cells, knockdown of ER&bgr; expression partially eliminated the blockade of DHT-induced cell proliferation. Conclusion: This study demonstrates that genistein may be a potential agent for prostate cancer therapy since genistein inhibits DHT-induced LAPC-4 and LNCaP prostate cancer cell proliferation but not HAEC cell growth. 17&bgr; − estradiol completely blocked DHT-induced cell growth in HAECs while inhibiting LAPC-4 cell proliferation, accounting for the side-effect of cardiovascular in Androgen deprivation therapy of prostate cancer. DHT-induced LNCaP prostate cancer cell proliferation can’t be attenuated by 17&bgr; − estradiol, suggesting 17&bgr; − estradiol is inactive in treating metastasized prostate cancer. ER&bgr; played an important role in the modulation of androgen receptor actions.

V-002 ANTIHYPERTENSIVE EFFECT OF AMLODIPINE PLUS TELMISARTAN OR AMILORIDE IN MODERATE AND HIGH-RISK PATIENTS

Background To evaluate the short-term effect of amlodipine piustelmisartan or amiloride on reduction of blood pressure and adverse effect in hypertensive patients with moderate or high risk of cardiovascular event. Methods In this randomized, blind-end trial, 106 hypertensive patients met Inclusion criteria are enrolled. Patients were randomly assigned to A group (amlodipine 2.5 mg plus telmisartan 80 mg group) or B group (amlodipine 2.5 mg plus 1 tablet of amiloride group); Amlodipine 2.5 mg could be added if blood pressure beyond control at 4-week. Follow up 24 weeks. Primary efficacy parameter was reduction of blood pressure at 24-week. Physical and laboratory characteristics and side effects were recorded. Results Baseline systolic blood pressure was160.5 ± 16.5 mm Hg and diastolic blood pressure was 98.7 ± 9.7 mm Hg. After 2-week treatment, mean systolic blood pressure in group A and B was (151.5 ± 14.8) mm Hg and (144.4 ± 13.9) mm Hg, respectively (P < 0.05). Mean diastolic blood pressure in group A and B was reduced to (91.7 ± 9.6) mm Hg and (90.1 ± 9.4) mm Hg, respectively (p>0.05). Blood pressure control rate was 47.2% and 58.1% in group A and B (P < 0.05). After 24-week therapy, there were no significant differences on reduction of blood pressures (SBPs 24.3 ± 15.8 vs 26.8 ± 13.4, p > 0.05) (DBPs 15.2 ± 9.2 vs15.7 ± 9.4, p > 0.05) or blood pressure control rates (67.9% of vs 71.7%, p > 0.05) in two groups. Compared with A and B groups, both of them reported equivalent of adverse effects (7.6% of amiloridevs 9.4% of telmisartan, p > 0.05). Conclusion Amlodipine-based antihypertensive combination strategies achieved satisfactory blood pressure control in hypertensive patients with moderate or high cardiovascular risk. Yet, more predominant efficacy on the reduction of blood pressure and blood pressure control rates shows in the combination of amlodipine and amiloride at 2-week follow-up. Adverse effects and organ benefits beyond reducing blood pressure warrant longer clinical observation.

V-005 CYCLOSPORINE A CONCENTRATION AND ANTIHYPERTENSIVE EFFECT BETWEEN AMLODIPINE AND VALSRATAN REGIMEN IN POST TRANSPLANT HYPERTENSIVE PATIENTS TREATED WITH CYCLOSPORINE A

Background To evaluate the effect of amlodipine and valsartan on blood pressure control in renal transplant patients and to analyze the correlation between CYP3A5 or MDR1 genotype and the antihypertensive effect of these two regimens. Methods One hundred fifty renal transplant patients with stage 1 or 2 hypertension were enrolled in the trial. Patients wererandomly assigned to amlodipine or valsartan. Metoprolol was added if blood pressure was not under control after four weeks. Blood pressure and plasma levels of cyclosporin A (CsA) were monitored during the 24-week trial. CYP3A5 and MDR1 genotypes were determined using a PCR-RFLP method. Results The demographic features and the baseline blood pressure were similar between these two groups. During the 24-week trial, the reduction of SBPwas similar between the amlodipine and valsartan groups. However, the reduction of DBP was significantly greater in the amlodipine group compared to the valsartan group at 12, 16 and 24 weeks of treatment. The plasma level of CsA at 2 hours of medication (C2) was significantly higher in the amlodipine group than that in the valsartan group after 4 weeks of the trial. The reduction of DBP at 24 weeks was greater in the subjects with CYP3A5 *3/*3 variant than those with CYP3A5*1/*1 variant (−13.5 ± 1.9 mmHg vs. −8.7 ± 1.6 mmHg, P < 0.05). Conclusion The present study demonstrated that amlodipine produced a greater reduction of DBP than valsartan although both amlodipine and valsartan resulted in a satisfactory control of blood pressure in patients with renal transplantation. Administration of amlodipine significantly increased the plasma concentration of CsA, and its effects on blood pressure control and CsA concentration may associated with CYP3A5 genotype in these subjects.

Amlodipine pius telmisartan or amiloride for hypertension in moderate and high-risk patients: A 24-week observation

Objective: The aim of the present study was to evaluate the shortterm effect of amlodipine-based antihypertensive combination regimens on reduction of blood pressure and adverse effect in hypertensive patients with moderate or high risk of cardiovascular event. Methods: In this randomized, blinded trial, 106 hypertensive patients met the inclusion criteria and were enrolled. Patients were randomly assigned to A group (amlodipine 2.5 mg plus telmisartan 80 mg group) or B group (amlodipine 2.5 mg plus 1 tablet of amiloride group); amlodipine 2.5 mg could be added if blood pressure beyond control at 4 weeks. Follow up was 24 weeks. Primary efficacy parameter was reduction of blood pressure at 24 weeks. Physical and laboratory characteristics and side effects were recorded. Results: Baseline systolic blood pressure was 160.5±16.5 mm Hg and diastolic blood pressure was 98.7±9.7 mm Hg. After 2 weeks treatment, mean systolic blood pressure in group A and B was (151.5± 14.8)mm Hg and (144.4±13.9)mmHg, respectively (Pb0.05). Mean diastolic blood pressure in group A and B was reduced to (91.7±9.6) mm Hg and (90.1±9.4)mm Hg, respectively (PN0.05). Blood pressure control rate was 47.2% and 58.1% in group A and B (Pb0.05). After 24 weeks of therapy, there were no significant differences on reduction of blood pressures (SBPs 24.3±15.8 vs 26.8±13.4, PN0.05) (DBPs 15.2±9.2 vs15.7±9.4, PN0.05) or blood pressure control rates (67.9% of vs 71.7%, PN0.05) in two groups. Compared with A and B groups, both of them reported equivalent of adverse effects (7.6% of amiloride vs 9.4% of telmisartan, PN0.05). Conclusion: Amlodipinebased antihypertensive combination strategies achieved satisfactory blood pressure control in hypertensive patients with moderate or high cardiovascular risk. Yet, more predominant efficacy on the reduction of blood pressure and blood pressure control rates shows in the combination of amlodipine and amiloride at 2-week follow-up. Adverse effects and organ benefits beyond reducing blood pressure warrant longer clinical observation.