Effects of atorvastatin on cardiac myocyte hypertrophy of neonatal rat induced by angiotensinⅡ in vitro and TLR4 expression

AIM: To assess effects of atorvastatin (Ator) on cardiac myocytes (CM) hypertrophy of neonatal rat induced by angiotensinⅡ (AngⅡ) in vitro and Toll like receptor 4 (TLR4) expression for theoretical bases of preventing and treating myocardial hypertrophy.METHODS: CM of neonatal Sprague-Dawley (SD) rats were isolated with trypsin digestion method and those growth-arrested cells were stimulated with 10^-7 mol/L AngⅡ in the presence of various concentrations of Ator.The method of coomassie brilliant blue was adopted to evaluate the protein contents of CM.The changes in β-MHC,AT₁ receptor and TLR4 mRNA expression were observed by RT-PCR.RESULTS: ① AngⅡ increased the protein content of CM and β-MHC mRNA expression significantly,upregulated AT₁ receptor and TLR4 gene expression.② In a dose-dependent manner,Ator inhibited the increases in the protein contents of CM and β-MHC mRNA expression induced by AngⅡ.③ In a dose-dependent manner,Ator downregulated the AT₁ receptor and TLR4 mRNA expression of CM hypertrophy of neonatal rat induced by AngⅡ.CONCLUSION: Ator inhibits CM hypertrophy,downregulates the AT₁ receptor and TLR4 gene expression.     

双氢睾酮诱导大鼠心肌细胞肥大模型的建立

应用胰蛋白酶分次消化法分离乳鼠心肌细胞,以差速贴壁法纯化心肌细胞,α-sarconme-actin抗体免疫细胞化学染色鉴定心肌细胞。心肌细胞在无血清无酚红培养基中培养48h后,用双氢睾酮(DHT)诱导心肌细胞肥大,建立心肌细胞肥大模型。24h后检测心肌细胞肥大的指标心肌细胞表面积;BCA法测定心肌细胞蛋白含量;半定量RTPCR两步法检测心肌细胞肥大的特征性基因—心房利钠因子(atrial natriuretic factor,ANF,β-肌球蛋白重链(β-myosin heavy chain,β-MHC)mRNA的表达。结果显示免疫细胞化学染色显示培养的心肌细胞纯度达到90%以上,心肌细胞分离良好。与对照组相比,10-8 mol/L的DHT能显著的增加心肌细胞表面积、蛋白质含量、ANP和β-MHC基因表达的增加(P0.01),心肌细胞肥大模型建立成功。     

Effects of Dryk1A,ASF and alternative splicing of CaMKⅡδ on myocardial hypertrophy in renovascular hypertensive rats

AIM: To investigate the role of dual-specificity tyrosine phosporylation-regulated kinase 1A (Dyrk1A)-alternative splicing factor (ASF)-calcium/calmodulin-dependent protein kinase Ⅱδ (CaMK Ⅱδ) pathway in the progression of myocardial hypertrophy in renovascular hypertensive rats. METHODS: The renovascular hypertension was induced by two-kidney one-clip (2K1C) method. The changes of blood pressure and myocardial hypertrophy were measured. The techniques of RT-PCR and Western blotting were used to detect CaMKⅡδ alternative splicing and the protein expression of Dyrk1A and ASF, respectively. RESULTS: Eight weeks after operation, systolic blood pressure (SBP) and diastolic blood pressure (DBP) in 2K1C rats increased (P<0.05). The increases in left ventricular weight (LVW), the ratio of LVW to body weight (BW) and the area of myocardial cells indicated that the hypertensive rats developed significant cardiac hypertrophy. The protein expression of Dyrk1A and mRNA expression of CaMKⅡδA and δB were significantly increased, while the protein expression of ASF and mRNA expression of CaMKⅡδC were decreased compared with sham-operated control rats (P<0.05). Treatment with Dryk1A inhibitor epigallocatechin gallate (EGCG) or harmine effectively attenuated cardiac hypertrophy and reversed the changes in the protein expression of Dyrk1A, ASF and alternative splicing of CaMKⅡδ (all P<0.05). CONCLUSION: Dyrk1A-ASF-CaMKⅡδ pathway plays a role in the development of myocardial hypertrophy in renovascular hypertensive rats.     

Influence of irbesarten on renal hypertrophy in streptozotocin-induced diabetic rats LIU Bi-cheng,LUO Dong-dong,ZHANG Xiao-liang.

AIM: To investigate the influence of irbesartan (Irb), a new angiotensin II receptor 1 antagonist, on renal hypertrophy in streptozotocin (STZ)-induced diabetic rats. METHODS: Sprague-Dawley (SD) rats were randomly divided into three groups: normal group (Group N, n=7), diabetic group (Group DN, n=6) and irbesartan treated group (Group DNI, n=7). In the experimental group, after the rats subjected to uninephrectomy, STZ was given by peritoneally injection at bolus dose of 50 mg/kg to induce diabetes. Blood glucose (BG), body weight (BW), urinary albumin excretion (Ualb), 24 hour proteinuria (24 h Upro) were measured at week 4, 8, 12, respectively. By the end of experiment at week 12, creatinine clearance (Ccr), kidney weight (KW), indicator of renal hypertrophy (KW/BW), renal total protein content (RTP), glomerular area (AG) and glomerular volume (VG) as well as glomerular basement membrane (GBM) were determined by semi-quantitative pathology technique. RESULTS: It was showed that there was no significant difference in BG between group DN and DNI, while Irb significantly reduced the increasing of Ualb, 24 h Upro in diabetic rats compared to control group (P＜0.01, respectively). Furthermore, Irb markedly inhibited the increasing of KW, KW/BW, RTP, A_G, V_G in diabetic rats (P＜0.05, P＜0.01, respectively). Of interest, Irb significantly prevented the increasing of GBM in diabetic rats. CONCLUSION: Irb exerts its early renal protective action by reducing proteinuria and inhibiting renal hypertrophy as well as the thickening of GBM.     

Effects of unilateral ovariectomy on recruitment and growth of follicles in the rainbow trout,Oncorhynchus mykiss

Virgin female rainbow trout, Oncorhynchus mykiss, were unilaterally ovariectomised at various stages of ovarian development to investigate the effect of the removal of one ovary on subsequent recruitment and growth of follicles in the remaining ovary. The right ovary was removed from groups of 12–15 fish, 12, 7 and 4 months before they were due to ovulate, and the gonadosomatic index and follicle number and size determined just prior to ovulation. There were no differences in fecundity or follicle size in fish unilaterally ovariectomised at 12 and 7 months prior to ovulation compared to the controls. However, in the females unilaterally ovariectomised 4 months prior to ovulation, the remaining ovary either had the normal number of follicles for a single ovary but of a significantly larger size than follicles in the controls, or alternatively had almost 70% more than the normal number of vitellogenic follicles but comprising two distinctly different size populations. Differences in plasma oestradiol-17β concentrations at the final sample were seen only in females unilaterally ovariectomised 4 months prior to ovulation, where the levels were significantly lower than both the sham operated and control fish (p < 0.05). These data show that in the rainbow trout, complete compensatory ovarian hypertrophy following unilateral ovariectomy can occur throughout a major part of ovarian development, but that follicle recruitment is limited to stages up to (and therefore fecundity is determined by) mid-vitellogenesis.     

长期服用氨氯地平对高血压病降压的疗效及左室结构功能的影响

目的；观察氨氯地平治疗高血压病的长期降压效应及左室结构功能的改变。方法：２６例患者日服氨氯地平５－１０ｍｇ，平均２４个月。于服药前，服药后４周及治疗结束时进行偶测血压，动态血压及彩色多普勒超声心动图检查。结果：治疗后偶测血压，白昼及夜间动态血压明显平稳下降，心率则差异无统计学意义，室间隔，左室后壁厚度及左室重量指数均显著下降。     

犬前列腺疾病概述

犬前列腺疾病是临床常见的公犬泌尿生殖道疾病,通常发生于老龄公犬,特别是6岁以上的未去势公犬。常见的犬前列腺疾病主要有前列腺炎、良性前列腺增生、前列腺囊肿、前列腺脓肿和前列腺癌。前列腺疾病的临床表现相似,表现为排尿困难、尿淋漓、血尿、前列腺液成分改变、便秘等,因此仅从病史、临床症状方面难以对疾病进行确诊,需要借助于X光检查、B超检查、前列腺液检查、组织/细胞学检查等特殊的诊断方法。在确诊的基础上根据不同的发病原因采取抗生素治疗,去势,前列腺切除手术等治疗方法。论文对各种常见犬前列腺疾病的病因、临床表现、诊断和治疗方案进行综述。     

Efficient generation of CLPG1‐edited rabbits using the CRISPR/Cas9 system

The sheep callipyge (CLPG) phenotype, a well‐known muscular hypertrophy syndrome, is caused by an A‐to‐G transition in the CLPG1 locus. The mechanisms of CLPG phenotype are very complicated and remain to be further studied. Lacking suitable animal models containing CLPG mutations may partially contribute to these unanswered mechanisms. In this study, we confirmed that the CLPG1 locus, especially the 12‐bp CLPG1 motif, is conserved in mammalian animals including rabbit. Then, we generated seven CLPG1‐edited rabbits with 100% efficiency using CRISPR/Cas9 system combined with cytoplasm injection technology. All the newborn rabbits were mosaicism with numerous kinds of mutations around the target sites. Among the nine screened potential off‐target sites (POTs) for the two sgRNAs used in this study, none off‐target effect was detected. This indicated that we efficiently and precisely generated CLPG1‐edited rabbits, and we believe that these newly generated rabbits will do help to unravel the mechanisms of the CLPG phenotype in the future.     

Changes of aryl hydrocarbon receptor in cardiac hypertrophy induced by high glucose in vitro

AIM: To investigate the changes of aryl hydrocarbon receptor (AhR) in the process of cardiomyocyte hypertrophy induced by high glucose, and to explore its potential mechanisms. METHODS: The rat cardiomyocytes (H9c2 cells) were divided into normal glucose group, high glucose group, DMSO group and resveratrol (an AhR antagonist) group. The content and distribution of AhR were observed with immunofluorescence staining. The myocardial cells were stained with rhodamine-labeled phalloidin to visualize cytoskeleton, and the cell surface area were determined after imaging by fluorescence microscopy. The generation of reactive oxygen species (ROS) in the cardiomyocytes was measured using a fluorescent probe DCFH-DA. The mRNA expression of AhR, CYP1A1, atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) were evaluated by real-time quantitative PCR (RT-qPCR). The protein levels of AhR, CYP1A1, ANP and BNP were assessed by Western blot. RESULTS: AhR was constitutively presented in the cytosol under normal-glucose condition and was translocated to the nuclei under high-glucose condition. High glucose induced cardiac hypertrophy, and increased ROS generation. Significant reductions in the cell size and ROS generation were observed after treated with resveratrol. The expression of AhR, CYP1A1, ANP and BNP at mRNA and protein levels in high glucose group was increased as compared with normal glucose group and resveratrol group, and the above-mentioned indexes significantly decreased in resveratrol group as compared with DMSO group. CONCLUSION: High glucose-induced cardiac hypertrophy increases AhR expression, which may be involved in the maintenance of glucose homeostasis in the cardiomyocytes. AhR translocation to the nucleus induced by high glucose results in the increases in CYP1A1 expression and ROS generation, which may be an important mechanism of high glucose-induced cardiomyocyte hypertrophy.     

Muscular hypertrophy and growth‐promoting effects in juvenile pejerrey (Odontesthes bonariensis) after oral administration of recombinant homologous growth hormone obtained by a highly efficient refolding process

Growth hormone (GH) can be orally administrated to fish in order to increase growth rates. Fish growth is characterized by the hyperplasia and hypertrophy of muscle fibre throughout adult life. In this respect, GH could affect directly and indirectly (by growth and metabolic factors) the development and growth of muscle fibres. Recombinant pejerrey GH (r‐pjGH) was expressed in Escherichia coli and refolded in a highly efficient batch dilution system, obtaining 0.1 g L^?1 of hormone without protein precipitation during the refolding procedure. Orally administered hormone to pejerrey produced a 30% increase in mean weight and stimulated liver insulin‐like growth factor type I (IGF‐I) mRNA expression after 1 month of treatment. Histological analyses showed that muscle growth was generated mainly by hypertrophy of the fibres. A higher r‐pjGH dose increased muscle fibre hypertrophy but somatic growth was negatively affected probably due to a reduced capacity of generating new fibres.