Intervention of Tongxinluo Capsule (通心络胶囊) against Vascular Lesion of Atherosclerosis and Its Effect on Lectin-like Oxidized Low Density Lipoprotein Receptor-1 Expression in Rabbits

Objective.- To investigate the prevention by Tongxinluo capsule （通心络胶囊, TXL） of vascular lesions and its effect on the levels of protein and gene expression of lectin-like oxidized low-density lipoprotein receptor-1 （LOX-1） of vascular wall in rabbits with atherosclerosis （AS）, and to explore its possible mechanism against AS. Methods： AS models were established by feeding New Zealand white rabbits with high-cholesterol diet, and 24 immature rabbits were randomly divided into the control group, model group and treated group （treated with TXL capsule）. The indexes of total cholesterol （TO） and low density lipoprotein （LDL） levels were measured at the 16th week. The intima thickness and the plaque area of abdominal aorta were quantitatively analyzed by pathological morphological analysis, the expression of macrophage and smooth muscle cell （SMC） in intima were detected by immunohistochemical method and histologic segments were stained by Hematoxilin-Eosin （HE） to identify the degree of atherosclerotic lesion in the model group and the prevention by TXL. The LOX-1 gene and protein expression in abdominal aorta was detected by semi-quantitative RT-PCR and immunohistochemistry, respectively. Results： In the model group, the levels of TC and LDL were significantly elevated, aortic intima thickened extensively, the intima area enhanced, and macrophages expression increased; the levels of LOX-1 gene and protein expression was up-regulated in endothelium and neo-intima of the abdominal aorta. The treatment with TXL reduced blood lipids, attenuated arterial intimal proliferation, markedly inhibited the expression of macrophage and excessively expressed the level of LOX-1. Conclusion：TXL has an inhibitory effect on blood lipids, and it can prevent the occurrence of vascular lesion and cure its development, and its protection against AS was possibly associated with a crucial endothelial protective action through lowering the expression of LOX-1 in vascular walls.

Shh、Gli1在脂多糖致小鼠急性肺损伤中的表达及与NF-κB的关系

目的探讨Sonic hedgehog(SHH)信号通路中Shh、Gli1及核因子-κB(nuclear factor kappa B,NF-κB)在脂多糖诱导的小鼠急性肺损伤中的表达变化。方法脂多糖腹腔注射复制小鼠急性肺损伤模型,实验动物随机分为实验组(脂多糖5 mg/kg)和对照组。分别于给药后6、12、24 h取肺组织,观察肺组织病理改变、肺湿干重比值(W/D)、RT-PCR和Western blot检测Shh、Gli1的m RNA和蛋白表达,Western blot检测NF-κB的蛋白表达。结果实验组肺组织病理损伤评分及W/D比值明显高于对照组(P<0.05)。Shh的m RNA表达水平在各时间点均高于对照组(P<0.05),且与时间呈正相关(P<0.01)。6 h时Gli1的m RNA表达水平与对照组无明显差异(P>0.05),12、24 h时明显高于对照组(P<0.01),且与时间呈正相关(P<0.01)。与对照组相比,实验组6、12、24 h时Shh、Gli1的蛋白表达水平均明显升高(P<0.05),且与时间呈正相关(P<0.01)。NF-κB的蛋白表达在各时间点均明显高于对照组(P<0.05)。结论脂多糖诱导的小鼠急性肺损伤中Shh、Gli1表达上调可能是脂多糖通过调控NF-κB来实现的。

Effects of benazepril on renal function and kidney expression of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in diabetic rats

Background Excessive deposition of extraceUular matrix （ECM） in the kidney is the hallmark of diabetic nephropathy. Increased matrix synthesis has been well documented but the effects of diabetes on degradative pathways, particularly in the in vivo setting. The renal protective effect of these pathways on matrix accumulation has not been fully elucidated. The present study was understaken to investigate the activity of matrix metalloproteinase-2 （MMP-2）, the expression of MMP-2 and tissue inhibitor of metalloproteinase-2 （TIMP-2） in kidney tissues of diabetic rats, and to explore the degradative pathway of type Ⅳ collagen （Ⅳ-C） and the renal protective effects of ACE inhibition- benazepril.

Paeoniflorin Improves Regional Cerebral Blood Flow and Suppresses Inflammatory Factors in the Hippocampus of Rats with Vascular Dementia

Objective： To explore the delayed neuroprotection induced by paeoniflorin（PF）, the principal component of Paeoniae radix prescribed in Chinese medicine, and its underlying mechanisms in rats subjected to vascular dementia（VD）. Methods： A rat model of VD was induced by bilateral common carotid arteries occlusion（BCCAO）. Low-dose or high-dose PF（20 or 40 mg/kg once per day） was administrated for 28 days after VD. The behavioral analysis of rat was measured by water morris. Regional cerebral blood volume（r CBV）, regional cerebral blood flow（r CBF） and mean transit time（MTT） were measured in the bilateral hippocampus by perfusion-weighted imaging（PWI）. The levels of interleukin-1β（IL-1β）, interleukin-6（IL-6）, and tumor necrosis factor alpha（TNF-α） were measured by commercially available enzyme-linked immunosorbent assay kits. Protein levels were evaluated by western blot analysis. m RNA levels were evaluated by real time-polymerase chain reaction. Western blotting was used to estimate p65 translocation. Results： The behavioral analysis showed that PF could decrease the escape latency time（P〈0.05）, and increase the residence time of the original platform quadrant and the across platform frequency in water maze in VD rats（P〈0.05）. Likewise, PF remarkably promoted the r CBV（P〈0.05）, r CBF and decreased per minute MTT（P〈0.05） in hippocampus of VD rats. Furthermore, PF decreased the release of IL-1β, IL-6 and TNF-α as well as inhibited the m RNA expression of IL-1β, IL-6 and TNF-α in the hippocampus of VD rats（P〈0.05 or P〈0.01）. PF also could decrease the protein expressions of inducible nitric oxide synthase and cyclooxygenase-2 in the hippocampus of VD rats（P〈0.05 or P〈0.01）. In addition, PF significantly inhibited the nuclear factor κB（NF-κB） pathway in the hippocampus of VD rats. Conclusions： PF significantly attenuates cognitive impairment, improves hippocampus perfusion and inhibits inflammatory response in VD rats. In addition, the anti-inflammatory effects of PF might be due to inhibiting the NF-κB pathway. PF may be a potential clinical application in improving VD.

Correlation of Gut Microbiome Between ASD Children and Mothers and Potential Biomarkers for Risk Assessment

Variation of maternal gut microbiota may increase the risk of autism spectrum disorders(ASDs) in offspring. Animal studies have indicated that maternal gut microbiota is related to neurodevelopmental abnormalities in mouse offspring, while it is unclear whether there is a correlation between gut microbiota of ASD children and their mothers. We examined the relationships between gut microbiome profiles of ASD children and those of their mothers, and evaluated the clinical discriminatory power of discovered bacterial biomarkers. Gut microbiome was profiled and evaluated by 16S ribosomal RNA gene sequencing in stool samples of 59 mother–child pairs of ASD children and 30 matched mother–child pairs of healthy children. Significant differences were observed in the gut microbiome composition between ASD and healthy children in our Chinese cohort. Several unique bacterial biomarkers, such as Alcaligenaceae and Acinetobacter, were identified. Mothers of ASD children had more Proteobacteria, Alphaproteobacteria, Moraxellaceae, and Acinetobacter than mothers of healthy children. There was a clear correlation between gut microbiome profiles of children and their mothers; however, children with ASD still had unique bacterial biomarkers, such as Alcaligenaceae, Enterobacteriaceae, and Clostridium. Candidate biomarkers discovered in this study had remarkable discriminatory power. The identified patterns of mother–child gut microbiome profiles may be important for assessing risks during the early stage and planning of personalized treatment and prevention of ASD via microbiota modulation.

Facile discovery of red blood cell deformation and compromised membrane/skeleton assembly in Prader—Willi syndrome

Prader—Willi syndrome(PWS)is a rare congenital disease with genetic alterations in chromosome 15.Although genetic disorders and DNA methylation abnormalities involved in PWS have been investigated to a significant degree,other anomalies such as those in erythrocytes may occur and these have not been clearly elucidated.In the present study,we uncovered slight anemia in children with PWS that was associated with increased red blood cell(RBC)distribution width(RDW)and contrarily reduced hematocrit(HCT)values.Intriguingly,the increased ratio in RDW to HCT allowed sufficient differentiation between the PWS patients from the healthy controls and,importantly,with individuals exhibiting conventional obesity.Further morphologic examinations revealed a significant deformity in erythrocytes and mild hemolysis in PWS patients.Comprehensive mechanistic investigations unveiled compromised membrane skeletal assembly and membrane lipid composition,and revealed a reduced F-actin/G-actin ratio in PWS patients.We ascribed these phenotypic changes in erythrocytes to the observed genetic defects,including DNA methylation abnormalities.Our collective data allowed us to uncover RBC deformation in children with PWS,and this may constitute an auxiliary indicator of PWS in early childhood.

Dynamic changes in myocardial matrix metalloproteinase activity in mice with viral myocarditis

Background Matrix metalloproteinases (MMPs) are the major regulators of collagen degradation involved in the pathogenesis of several diseases of the heart The purpose of this study was to investigate the dynamic changes in myocardial MMP activity in mice with viral myocarditis (VM), the relationship between MMP activity and both cardiac function and the quantity of myocardial collagen, and the role MMPs playing in the pathological lesions of VM KH*2/5DMethods Sixty-five six-week-old male DBA/2 mice were divided into two groups Mice in the infected group (n=50) were inoculated intraperitoneally with 0 14 ml of Coxsackievirus B 3 (CVB 3, Nancy strain) Control mice (n=15) were inoculated intraperitoneally with 0 14 ml of Eagle’s medium Eight infected mice and three control mice were sacrificed on each of days 3, 7, 10, 21 and 30 after inoculation MMP activity was measured on an SDS-PAGE substrate gel embedded with type Ⅰ gelatin (zymography) Echocardiographic studies were performed under anesthesia with 3% chloralhydrate administered intraperitoneally (0 01 ml/g-0 015 ml/g) Cardiac systolic function indices, such as peak velocity of the aorta (V p), flow velocity integral of the aorta (V i), ejection fraction (EF), and fractional shortening (FS) were determined by echocardiography Histological cross sections of the hearts were stained with hematoxylin-eosin and myocardial histopathological scores were determined under an optical microscope The amount of myocardial collagen was measured by means of hydroxyproline quantification Results In virus-infected mice, both MMP-2 and MMP-9 activities were significantly higher than in control mice, reaching a peak on day 10 ( P <0 01) On day 10, cardiac systolic function indices (EF, FS, V p, and V i) were all significantly lower compared both to other stages following viral inoculation and to the control group ( P <0 05) In the acute stage, the amount of myocardial collagen in mice with VM was not significantly different from normal control mice ( P >0 05) However, the amount of myocardial collagen in infected mice at the recovery stage (on days 21 and 30) was significantly greater than those of the control mice MMP-2 and MMP-9 activities positively correlated with myocardial histopathological scores ( r =0 801,0 821, P <0 01) and negatively correlated with V p ( r =-0 649, -0 683, P <0 01) and V i ( r =-0 711, -0 755, P <0 01) However, V p negatively correlated with myocardial histopathological scores ( r =-0 756, P <0 01) Conclusions In mice with VM, the activities of myocardial MMP-2 and MMP-9 increase significantly during the acute stage, and the total quantity of myocardial collagen increases by the time of recovery These changes are associated with myocardial interstition remodeling and cardiac dysfunction MMP activity is an important reference marker for myocardial pathological lesions and can be used to evaluate the severity of myocardial interstitial damage and cardiac dysfunction

Tong-xin-luo capsule inhibits left ventricular remodeling in spontaneously hypertensive rats by enhancing PPAR-γ expression and suppressing NF-κB activity

Background Tong-xin-luo capsule （TXL）, used as a traditional Chinese herb, offeres a therapeutic potential for treatment of cardiovascular diseases. It has been shown to exert a variety of pharmacological effects, including antihypertensive effects, and is able to improve ventricular remodeling. However, the mechanisms of its action are not completely understood. The aim of this study was to evaluate the molecular mechanisms of Tong-xin-luo capsule on left ventricular remodeling in spontaneously hypertensive rats （SHR）. Methods Sixteen eight-week-old SHRs were randomized into an SHR group （n=8） and a TXL group （n=8） that were given Tong-xin-luo capsule （1.5 mg·kg^-1·d^-1）. Eight Wistar Kyoto （WKY） rats fed with 0.9% NaCl served as the control group （WKY group）. Systolic blood pressure （BP）, body weight and heart rate were monitored once every two weeks. Ventricular remodeling was detected by histopathological examination. Nuclear factor kappa B P65 （NF-κB P65） and peroxisome proliferators activated receptor y （PPAR-γ） protein and phosphorylated inhibitor kappa a （IκBα） protein were detected by immunohistochemistry and western blot respectively. The physical interaction of the P65-P50 heterodimer with IκBα and NF-κB were measured by co-immunoprecipitation. PPAR-γ mRNA, collagen Ⅰ mRNA and collagen Ⅲ mHNA were measured by real-time PCR.Results TXL inhibited NF-κB P65 expression and ventricular remodeling and suppressed the activation of NF-κB compared with the SHR group （P〈0.01, P〈0.05）. TXL reduced IκBα phosphorylation, increased expression of PPAR-γ protein and enhanced the physical interaction of the P65-P50 heterodimer with IκBα. The mRNA expression of PPAR-γ was enhanced but the mRNA expression of collagen Ⅰ mRNA and collagen Ⅲ mRNA were suppressed by TXL. Conclusions In spontaneously hypertensive rats, TXL could inhibit ventricular remodeling induced by hypertension, and the inhibitory effect might be associated with the process of TXL increasing the expression of PPAR-γ that could result in the inhibition of the activation of NF-κB.