旨在探究酪氨酸相关蛋白酶1(tyrosinase related protein 1,TYRP1)对香猪原代表皮黑素细胞黑色素生成的影响。本研究选用具有“两头乌”毛色特征的3月龄健康香猪3头,每头猪采取头部黑色皮肤组织和背部白色皮肤组织。通过HE染色观察不同...旨在探究酪氨酸相关蛋白酶1(tyrosinase related protein 1,TYRP1)对香猪原代表皮黑素细胞黑色素生成的影响。本研究选用具有“两头乌”毛色特征的3月龄健康香猪3头,每头猪采取头部黑色皮肤组织和背部白色皮肤组织。通过HE染色观察不同颜色香猪皮肤的毛囊结构及黑素细胞的分布特征;体外培养香猪原代表皮黑素细胞并通过多巴(L-Dopa)染色、实时荧光定量PCR和Western Blot方法进行鉴定;根据TYRP1基因序列构建5条siRNA,采用实时荧光定量PCR筛选出干扰效率最高的siRNA进行转染。成功干扰黑素细胞内TYRP1的表达后,分别通过实时荧光定量PCR、Western Blot、黑色素含量检测方法对黑素细胞内TYR、TYRP1及TYRP2的mRNA与蛋白相对表达量、细胞内总黑色素含量进行检测。结果显示,在香猪黑色和白色皮肤中都观察到完整的毛囊结构,黑素细胞主要分布在香猪黑色被毛皮肤中的毛囊外根鞘部位以及表皮。在MelM培养基作用下,黑素细胞生长旺盛,细胞形态、生长曲线、多巴染色、实时荧光定量PCR及Western Blot结果均表明培养的黑素细胞维持正常的生物学特性。敲降TYRP1后下调香猪表皮黑素细胞中黑色素生成相关基因TYR、TYRP1和TYRP2的mRNA和蛋白的表达,同时对黑色素的形成具有抑制作用。TYRP1基因能够影响香猪表皮黑素细胞黑色素的生成,研究结果可为探索TYRP1基因对香猪黑色素沉积的分子机制提供试验参考和基础数据。展开更多
BACKGROUND In recent years,studies have found that the occurrence and development of diabetic cardiomyopathy(DCM)is closely related to an increase in polyadenosine diphosphate-ribose polymerase-1(PARP-1)activity.PARP-...BACKGROUND In recent years,studies have found that the occurrence and development of diabetic cardiomyopathy(DCM)is closely related to an increase in polyadenosine diphosphate-ribose polymerase-1(PARP-1)activity.PARP-1 activation could be involved in the pathophysiological process of DCM by promoting oxidative stress,the inflammatory response,apoptosis and myocardial fibrosis.AIM To investigate the mechanism of liraglutide in improving myocardial injury in type 2 diabetic rats,further clarified the protective effect of liraglutide on the heart,and provided a new option for the treatment of DCM.METHODS Forty healthy male SD rats aged 6 wk were randomly divided into two groups,a normal control group(n=10)and a model group(n=30),which were fed an ordinary diet and a high-sugar and high-fat diet,respectively.After successful modeling,the rats in the model group were fed a high-glucose and high-fat diet for 4 wk and randomly divided into a model group and an intervention group(further divided into a high-dose group and a low-dose group).The rats were fed a high-glucose and high-fat diet for 8 wk and then started drug intervention.Blood samples were collected from the abdominal aorta to detect fasting blood glucose and lipid profiles.Intact heart tissue was dissected,and its weight was used to calculate the heart weight index.Haematoxylin and eosin staining was used to observe the pathological changes in the myocardium and the expression of PARP-1 in the heart by immunohistochemistry.RESULTS The body weight and heart weight index of rats in the model group were significantly increased compared with those in the normal control group,and those in the intervention group were decreased compared with those in the model group,with a more obvious decrease observed in the high-dose group(P<0.05).In the model group,myocardial fibers were disordered,and inflammatory cells and interstitial fibrosis were observed.The cardiomyopathy of rats in the intervention group was improved to different degrees,the myocardial fibers were arranged neatly,and the myocardial cells were clearly striated;the improvement was more obvious in the high-dose group.Compared with the normal control group,the expression of PARP-1 in myocardial tissue of the model group was increased,and the difference was statistically significant(P<0.05).After liraglutide intervention,compared with the model group,the expression of PARP-1 in myocardial tissue was decreased,and the reduction was more obvious in the high-dose group(P<0.05)but still higher than that in the normal control group.CONCLUSION Liraglutide may improve myocardial injury in type 2 diabetic rats by inhibiting the expression of myocardial PARP-1 in a dose-dependent manner.展开更多
[Objectives]To investigate the spasticity of rat tail and the expression of dopamine receptor-1(DRD1)mRNA in the spinal cord after spinal cord injury(SCI)induced tail spasticity in rats.[Methods]Adult male Wistar rats...[Objectives]To investigate the spasticity of rat tail and the expression of dopamine receptor-1(DRD1)mRNA in the spinal cord after spinal cord injury(SCI)induced tail spasticity in rats.[Methods]Adult male Wistar rats were randomly divided into Sham group and SCI group.The second sacral spinal cord(S2)segment of SCI rats was completely transected.60 d after operation,the rat tail spasticity was scored,and then the spinal cord tissues below the level of S2 spinal cord transection were taken.The expression of DRD1 mRNA in the sacrococcygeal spinal cord was detected by qPCR.In addition,3 normal rats were used for DAR/neuronal nuclei(NeuN)and DRD1/choline acetyltransferase(ChAT)immunofluorescence staining to study the distribution of DRD1 in spinal cord and the properties of DRD1 positive cells.[Results]60 d after operation in SCI group,the tail spasticity of rats developed fully,and the symptoms of spasticity were typical.qPCR results showed that the expression of DRD1 mRNA in SCI group was significantly lower than that in Sham group(P<0.05).DRD1 was widely distributed in the dorsal horn,intermediate zone and ventral horn at the sacrococcygeal end of the rat spinal cord.[Conclusions]The decrease of DRD1 mRNA expression after SCI may be related to the occurrence and development of spasticity.展开更多
文摘旨在探究酪氨酸相关蛋白酶1(tyrosinase related protein 1,TYRP1)对香猪原代表皮黑素细胞黑色素生成的影响。本研究选用具有“两头乌”毛色特征的3月龄健康香猪3头,每头猪采取头部黑色皮肤组织和背部白色皮肤组织。通过HE染色观察不同颜色香猪皮肤的毛囊结构及黑素细胞的分布特征;体外培养香猪原代表皮黑素细胞并通过多巴(L-Dopa)染色、实时荧光定量PCR和Western Blot方法进行鉴定;根据TYRP1基因序列构建5条siRNA,采用实时荧光定量PCR筛选出干扰效率最高的siRNA进行转染。成功干扰黑素细胞内TYRP1的表达后,分别通过实时荧光定量PCR、Western Blot、黑色素含量检测方法对黑素细胞内TYR、TYRP1及TYRP2的mRNA与蛋白相对表达量、细胞内总黑色素含量进行检测。结果显示,在香猪黑色和白色皮肤中都观察到完整的毛囊结构,黑素细胞主要分布在香猪黑色被毛皮肤中的毛囊外根鞘部位以及表皮。在MelM培养基作用下,黑素细胞生长旺盛,细胞形态、生长曲线、多巴染色、实时荧光定量PCR及Western Blot结果均表明培养的黑素细胞维持正常的生物学特性。敲降TYRP1后下调香猪表皮黑素细胞中黑色素生成相关基因TYR、TYRP1和TYRP2的mRNA和蛋白的表达,同时对黑色素的形成具有抑制作用。TYRP1基因能够影响香猪表皮黑素细胞黑色素的生成,研究结果可为探索TYRP1基因对香猪黑色素沉积的分子机制提供试验参考和基础数据。
基金Supported by Shanxi Provincial Natural Science Foundation,No.201701D121159Shanxi Provincial Health and Family Planning Commission,No.2014016Health Commission of Shanxi Province,No.2019020.
文摘BACKGROUND In recent years,studies have found that the occurrence and development of diabetic cardiomyopathy(DCM)is closely related to an increase in polyadenosine diphosphate-ribose polymerase-1(PARP-1)activity.PARP-1 activation could be involved in the pathophysiological process of DCM by promoting oxidative stress,the inflammatory response,apoptosis and myocardial fibrosis.AIM To investigate the mechanism of liraglutide in improving myocardial injury in type 2 diabetic rats,further clarified the protective effect of liraglutide on the heart,and provided a new option for the treatment of DCM.METHODS Forty healthy male SD rats aged 6 wk were randomly divided into two groups,a normal control group(n=10)and a model group(n=30),which were fed an ordinary diet and a high-sugar and high-fat diet,respectively.After successful modeling,the rats in the model group were fed a high-glucose and high-fat diet for 4 wk and randomly divided into a model group and an intervention group(further divided into a high-dose group and a low-dose group).The rats were fed a high-glucose and high-fat diet for 8 wk and then started drug intervention.Blood samples were collected from the abdominal aorta to detect fasting blood glucose and lipid profiles.Intact heart tissue was dissected,and its weight was used to calculate the heart weight index.Haematoxylin and eosin staining was used to observe the pathological changes in the myocardium and the expression of PARP-1 in the heart by immunohistochemistry.RESULTS The body weight and heart weight index of rats in the model group were significantly increased compared with those in the normal control group,and those in the intervention group were decreased compared with those in the model group,with a more obvious decrease observed in the high-dose group(P<0.05).In the model group,myocardial fibers were disordered,and inflammatory cells and interstitial fibrosis were observed.The cardiomyopathy of rats in the intervention group was improved to different degrees,the myocardial fibers were arranged neatly,and the myocardial cells were clearly striated;the improvement was more obvious in the high-dose group.Compared with the normal control group,the expression of PARP-1 in myocardial tissue of the model group was increased,and the difference was statistically significant(P<0.05).After liraglutide intervention,compared with the model group,the expression of PARP-1 in myocardial tissue was decreased,and the reduction was more obvious in the high-dose group(P<0.05)but still higher than that in the normal control group.CONCLUSION Liraglutide may improve myocardial injury in type 2 diabetic rats by inhibiting the expression of myocardial PARP-1 in a dose-dependent manner.
基金Supported by Youth Project of National Natural Science Foundation of China(81501080)Funding Project for Introducing Overseas Students in Hebei Provincial Department of Human Resources and Social Security(C20190174)Construction of Chengde Biomedical Industry Research Institute(202205B086)。
文摘[Objectives]To investigate the spasticity of rat tail and the expression of dopamine receptor-1(DRD1)mRNA in the spinal cord after spinal cord injury(SCI)induced tail spasticity in rats.[Methods]Adult male Wistar rats were randomly divided into Sham group and SCI group.The second sacral spinal cord(S2)segment of SCI rats was completely transected.60 d after operation,the rat tail spasticity was scored,and then the spinal cord tissues below the level of S2 spinal cord transection were taken.The expression of DRD1 mRNA in the sacrococcygeal spinal cord was detected by qPCR.In addition,3 normal rats were used for DAR/neuronal nuclei(NeuN)and DRD1/choline acetyltransferase(ChAT)immunofluorescence staining to study the distribution of DRD1 in spinal cord and the properties of DRD1 positive cells.[Results]60 d after operation in SCI group,the tail spasticity of rats developed fully,and the symptoms of spasticity were typical.qPCR results showed that the expression of DRD1 mRNA in SCI group was significantly lower than that in Sham group(P<0.05).DRD1 was widely distributed in the dorsal horn,intermediate zone and ventral horn at the sacrococcygeal end of the rat spinal cord.[Conclusions]The decrease of DRD1 mRNA expression after SCI may be related to the occurrence and development of spasticity.
