OBJECTIVE:Morroniside (MOR) has been reported to ameliorate inflammation in cardiovascular and cerebrovascular disease; however, its impact and mechanism on rheumatoid arthritis (RA) remains unclear. This study aimed to investigate the beneficial role of morroniside in treating RA and explore the anti-invasive mechanism of morroniside on joint destruction. METHODS:In vitro (using primary rat articular fibroblast-like synoviocytes (FLSs)) a wound healing assay was used to detect the migration of primary rat FLSs. Quantitative RT-PCR was used to measure the transcription of matrix metalloproteinase (MMP) MMP2 and MMP9. Western blot was used to measure the expression of MMP2, MMP9, p65, phosphorylated-p65 (p-p65), inhibitor of nuclear factor (NF)-[Formula: see text]Bα (I[Formula: see text]Bα), I[Formula: see text]B kinase α/β (IKKα/β), and phosphorylated-IKKα/β. Immunofluorescence assay was used to measure the nuclear translocation of p65. In vivo (using rats with collagen-induced arthritis), the joint histopathological changes were detected by routine hematoxylin and eosin. Immunohistochemistry assay was used to measure the expression MMP2 and MMP9. RESULTS:Morroniside diminished tumor necrosis factor (TNF)α-stimulated migration of primary rat articular FLSs. Morroniside also attenuated RA-FLSs invasion into joint and joint destruction in rats with collagen-induced arthritis (CIA). Further analysis revealed that morroniside inhibited the overexpression of matrix metalloproteinase MMP2 and MMP9 in TNFα-stimulated primary rat articular FLSs and joints of CIA rats. Mechanistically, morroniside suppressed the activation of I[Formula: see text]B kinase α/β, which resulted in elevated levels of the inhibitor of nuclear factor (NF)-[Formula: see text]B. CONCLUSION:The present study suggested that morroniside can prevent joint destruction by suppressing the activation of the NF-[Formula: see text]B/MMPs pathway, thereby preventing FLSs invasion.

Background Angiopoietin-like protein 3 (ANGPTL3) is secreted by hepatocytes and inhibits lipoprotein lipase and endothelial lipase activity. Previous studies reported the correlation between plasma ANGPTL3 levels and high-density lipoprotein (HDL). Recently ANGPTL3 was found to preferentially bind to HDL in healthy human circulation. Here, we examined whether ANGPTL3, as a component of HDL, modulates HDL function and affects HDL other components in human and mice with non-diabetes or type 2 diabetes mellitus. Methods HDL was isolated from the plasma of female non-diabetic subjects and type-2 diabetic mellitus (T2DM) patients. Immunoprecipitation, western blot, and ELISA assays were used to examine ANGPTL3 levels in HDL. Db/m and db/db mice, AAV virus mediated ANGPTL3 overexpression and knockdown models and ANGPTL3 knockout mice were used. The cholesterol efflux capacity induced by HDL was analyzed in macrophages preloaded with fluorescent cholesterol. The anti-inflammation capacity of HDL was assessed using flow cytometry to measure VCAM-1 and ICAM-1 expression levels in TNF-α-stimulated endothelial cells pretreated with HDL. Results ANGPTL3 was found to bind to HDL and be a component of HDL in both non-diabetic subjects and T2DM patients. Flag-ANGPTL3 was found in the HDL of transgenic mice overexpressing Flag-ANGPTL3. ANGPLT3 of HDL was positively associated with cholesterol efflux in female non-diabetic controls (r = 0.4102, p = 0.0117) but not in female T2DM patients (r = − 0.1725, p = 0.3224). Lower ANGPTL3 levels of HDL were found in diabetic (db/db) mice compared to control (db/m) mice and were associated with reduced cholesterol efflux and inhibition of VCAM-1 and ICAM-1 expression in endothelial cells (p < 0.05 for all). Following AAV-mediated ANGPTL3 cDNA transfer in db/db mice, ANGPTL3 levels were found to be increased in HDL, and corresponded to increased cholesterol efflux and decreased ICAM-1 expression. In contrast, knockdown of ANGPTL3 levels in HDL by AAV-mediated shRNA transfer led to a reduction in HDL function (p < 0.05 for both). Plasma total cholesterol, total triglycerides, HDL-c, protein components of HDL and the cholesterol efflux function of HDL were lower in ANGPTL3−/− mice than ANGPTL3+/+ mice, suggesting that ANGPTL3 in HDL may regulate HDL function by disrupting the balance of protein components in HDL. Conclusion ANGPTL3 was identified as a component of HDL in humans and mice. ANGPTL3 of HDL regulated cholesterol efflux and the anti-inflammatory functions of HDL in T2DM mice. Both the protein components of HDL and cholesterol efflux capacity of HDL were decreased in ANGPTL3−/− mice. Our findings suggest that ANGPTL3 in HDL may regulate HDL function by disrupting the balance of protein components in HDL. Our study contributes to a more comprehensive understanding of the role of ANGPTL3 in lipid metabolism.

AbstractAims/IntroductionThe association between serum angiopoietin‐like 4 (ANGPTL4) levels and the severity of diabetic kidney disease (DKD) in patients with type 2 diabetes mellitus remains unclear.MethodsA total of 1,115 type 2 diabetes mellitus patients were analyzed in this cross‐sectional study. DKD index included DKD stages defined by estimated glomerular filtration rate, the albuminuria grades and DKD risk management grades. Serum levels of ANGPTL4 and other biomarkers were detected. Multivariable‐adjusted linear and logistic analyses were used to study the association between ANGPTL4 and DKD. The protein levels of ANGPTL4 were assessed in the kidney. Renal tubular cells were stimulated with glucose to study ANGPTL4 expression.ResultsCompared with the participants in the third or fourth quantile of ANGPTL4, those in the first or second quantile of ANGPTL4 were younger, with lower glycated hemoglobin, triglycerides and urinary albumin creatinine ratio (all P < 0.05). There was a negative nonlinear relationship between ANGPTL4 and estimated glomerular filtration rate in type 2 diabetes mellitus patients. One standard deviation increased serum ANGPTL4 levels, the odds ratio of having DKD was 1.40 (95% confidence interval 1.08–1.80). The mediation analysis showed that triglycerides did not mediate the association between ANGPTL4 and DKD. Furthermore, ANGPTL4 could be the strongest among multiple panels of biomarkers in its association of DKD. Compared with mice at 8 weeks‐of‐age, db/db mice at 18 weeks‐of‐age had increased ANGPTL4 expression in glomeruli and tubular segments. In vitro, glucose could stimulate ANGPTL4 expression in tubular cells in a dose‐dependent manner.ConclusionsANGPTL4 could be a potential marker and therapeutic target for DKD treatment.

Diabetic cardiomyopathy (DCM), a main cardiovascular complication of diabetes, can eventually develop into heart failure and affect the prognosis of patients. Myocardial fibrosis is the main factor causing ventricular wall stiffness and heart failure in DCM. Early control of myocardial fibrosis in DCM is of great significance to prevent or postpone the progression of DCM to heart failure. A growing body of evidence suggests that cardiomyocytes, immunocytes, and endothelial cells involve fibrogenic actions, however, cardiac fibroblasts, the main participants in collagen production, are situated in the most central position in cardiac fibrosis. In this review, we systematically elaborate the source and physiological role of myocardial fibroblasts in the context of DCM, and we also discuss the potential action and mechanism of cardiac fibroblasts in promoting fibrosis, so as to provide guidance for formulating strategies for prevention and treatment of cardiac fibrosis in DCM.

目的 探讨三碘甲腺原氨酸(T3)对甲状腺乳头状癌细胞TPC-1增殖的影响及其机制.方法 将TPC-1细胞分为对照组(0 ng/ml T3处理组)、12.5 ng/ml T3处理组和50 ng/ml T3处理组,分别用对应T3浓度处理细胞,实时无标记细胞功能分析技术(RTCA)和克隆形成实验检测细胞的增殖能力,Western blot检测细胞中PDZK1表达水平.利用慢病毒转染的方法将TPC-1细胞分为敲减PDZK1(shPDZK1)组、敲减对照(shNC)组、过表达PDZK1(PDZK1)组、过表达对照(Vector)组,采用Western blot检测TPC-1细胞中PDZK1表达水平,同时用不同浓度的T3(0、12.5、50 ng/ml)处理各组细胞,利用克隆形成实验检测细胞的克隆形成能力.结果 12.5及50 ng/ml T3处理组TPC-1的细胞指数及克隆形成数明显高于对照组,差异均有高度统计学意义(P<0.01).12.5及50 ng/ml T3处理组细胞中PDZK1表达量明显高于对照组,差异均有统计学意义(P<0.05或P<0.01).与shNC组比较,sh-PDZK1组细胞中PDZK1的表达量明显较低,差异有统计学意义(P<0.05).与Vector组比较,PDZK1组细胞PDZK1的表达量明显升高,差异有高度统计学意义(P<0.01).在0、12.5、50 ng/ml T3处理下,与shNC组比较,shPDZK1组细胞的克隆形成数均明显减少,差异均有统计学意义(P<0.05或P<0.01).在0、12.5、50 ng/ml T3处理下,与Vector组比较,PDZK1组细胞的克隆形成数均明显增加,差异均有统计学意义(P<0.05或P<0.01).结论 T3可能通过上调TPC-1细胞PDZK1的表达调控细胞增殖.

Background Obesity is an important risk factor for hyperuricemia. We aimed to explore the relationship between perirenal fat thickness (PrFT) and paranephric fat thickness (PnFT) and serum uric acid (SUA) in patients with type 2 diabetes mellitus (T2DM). Methods This was a cross-sectional study involving 257 patients with T2DM recruited from Beijing Luhe Hospital from September 2019 to May 2020. The basic and clinical information such as age, gender, duration of diabetes was collected through the medical records. All patients underwent a physical examination including height, weight, waist circumference, hip circumference, systolic blood pressures and diastolic blood pressure. The venous blood and urine samples were collected to measure SUA, fasting blood glucose, total cholesterol, triglyceride, low-density lipoprotein-cholesterol, high-density lipoprotein-cholesterol, serum creatinine, blood urea nitrogen and glycosylated hemoglobin. PrFT and PnFT were measured via ultrasonography. Pearson correlation test and linear regression analysis were used to analyze the association between PrFT and PnFT and SUA. Results We found that PrFT and PnFT increased according to the tertiles of SUA level ( P = 0.001 and P = 0.009, respectively). In addition, the PrFT and PnFT were positively associated with SUA level ( r = 0.25, P < 0.001, r = 0.23, P < 0.001, respectively). Moreover, this association was stronger in males, non-obesity patients and patients with normal renal function. In the multivariate analysis, the PrFT was independently associated with SUA level after adjusting confounding factors. Conclusions The PrFT was independently associated with SUA level in patients with T2DM.

Aim: Thyroid cancer (TC) is the most common malignant tumor of the endocrine system. Studies have showed that Triiodothyronine (T3) promotes the proliferation of papillary thyroid cancer (PTC) cells, but the specific mechanism remains unclear. Several studies have showed that PDZK1 played important roles in the occurrence and development of cancer. However, the biological function of PDZK1 in PTC remains unclear. Therefore, the aims of this study were to investigate the effect of PDZK1 in PTC and the underlying mechanism. Materials and Methods: The effect of T3 on proliferation of PTC cell (TPC-1) was analyzed by colony formation assay and real time cell analysis. Immunofluorescence staining aimed to analyze protein expression. RNA-seq was used to analyze the expression of PDZK1. Meanwhile, Western blot was used to verify the protein expression. The effect of PDZK1 on PTC cell proliferation was investigated by Cell Counting Kit-8(CCK8), real time cell analysis, flow cytometry and transwell assay, respectively. Results: We found that T3 increased the expression of PDZK1 in TPC-1 cells and promote the proliferation of TPC-1 cells which can be weakened after PDZK1 was knocked down. Immunofluorescence staining showed that the expression of PDZK1 was higher in PTC than paracancerous tissues. And the analysis of 6 benign thyroid nodules and 4 thyroid cancer tissues by RNA-Seq showed that the expression of PDZK1 was increased in PTC tissues, the expression of PDZK1 was also increased in PTC cells compared with the normal thyroid epithelium. In addition, PDZK1 promoted TPC-1 cell proliferation was detected by real-time cell analysis (RTCA) and CCK8. Flow cytometry analysis showed that PDZK1 increased cell cycle at S phase and decreased at G1 phase of TPC-1. PDZK1 promoted the invasion of TPC-1 cell was tested by transwell. Conclusion: These results suggest that T3 can promote the proliferation of PTC cells, which may be mediated by PDZK1. This study illustrates a possible mechanism by which T3 promotes the proliferation of PTC and provides a theoretical basis for the prevention and treatment of PTC.

目的 研究血清中血管生成素样蛋白4(angiopoietin-like 4,ANGPTL4)与非酒精性脂肪肝(non-alcoholic fatty liver disease,NAFLD)及严重程度的关系,探讨ANGPTL4用于诊断NAFLD的价值.方法 选择首都医科大学附属北京潞河医院2018年1月至12月诊断为NAFLD的患者52例为病例组,同期选择47例健康者为对照组.所有研究对象都由同一名超声科医生进行了腹部超声检查,并将NAFLD患者分为轻度、中度和重度.排除糖尿病、冠状动脉粥样硬化性心脏病(以下简称冠心病)、脑血管病、慢性肝病、服用降脂药或其他药物的患者.用酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)定量检测血清ANGPTL4的浓度,分析对照组与轻、中、重度NAFLD患者ANGPLT4的差异.以超声诊断为金标准,分析ANGPTL4作为NAFLD潜在的血清分子标志物的可能性.结果 与对照组相比,NAFLD患者血清ANGPTL4浓度升高,并随着NAFLD的严重程度有升高趋势(F=4.553,P=0.0354).ANGPTL4的受试者工作特征曲线(receiver operating characteristic curve,ROC)下面积为0.638,以最大约登指数确定NAFLD的诊断阈值,最佳临界值为90.93 ng/mL.以此标准诊断的结果与超声诊断结果进行比较,灵敏度=100％,特异度=46.8％,Kappa=0.48.结论 血清ANGPTL4浓度与NAFLD及其严重程度有关,用于NAFLD临床辅助诊断的误诊率较高.