Patatin-like phospholipase domain containing 5 （PNPLA5） is a neotype neutral lipase with dual activity of anabolism and catabolism in vitro and in vivo, which has a low mRNA expression level in humans and mice. PNPL...Patatin-like phospholipase domain containing 5 （PNPLA5） is a neotype neutral lipase with dual activity of anabolism and catabolism in vitro and in vivo, which has a low mRNA expression level in humans and mice. PNPLA5, which is localized to lipid droplets and required for efficient autophagy by optimal initiation, has been speculated to possess triglyceride hydro- lase activity, and has been associated with low density lipoprotein cholesterol （LDL-C）. Above all, PNPLA5 is a relatively new gene, which is reported less about its biological function research, especially the function research in the rats is still blank. In this study, we examined the spatiotemporal expression profile of PNPLA5 and found that it was expressed at low levels in most organs of Sprague Dawley （SD） rats, but was present at very high levels in the skin and testes. To furth.er determine the biological function of PNPLA5 in mammals, we generated PNPLA5-knockout SD rats using the clustered regularly-interspaced short palindromic repeats （CRISPR）/Cas9 system. PNPLA5-null rats were viable, but showed a variety of phenotypic abnormalities, such as abnormal bleeding, and varied hematobiochemical parameters including increased serum total cholesterol （TC）, tdglycerides and high density lipoprotein cholesterol （HDL-C） level, and reduced LDL-C level, compared with wild-type control rats. These data are consistent with an important role for PNPLA5 in lipid metabolism, provJdJng a new target gene and animal model for treatment of cardiovascular diseases in the future.展开更多
Background:Inflammation is a complex physiological and pathological process.Although many types of inflammation are well characterized,their physiological func-tions are largely unknown.tRNA aspartic acid methyltransf...Background:Inflammation is a complex physiological and pathological process.Although many types of inflammation are well characterized,their physiological func-tions are largely unknown.tRNA aspartic acid methyltransferase 1(TRDMT1)has been implicated as a stress-related protein,but its intrinsic biological role is unclear.Methods:We constructed a Trdmt1 knockout rat and adopted the LPS-induced sepsis model.Survival curve,histopathological examination,expression of inflammatory fac-tors,and protein level of TLR4 pathway were analyzed.Results:Trdmt1 deletion had no obvious impact on development and growth.Trdmt1 de-letion slightly increased the mortality during aging.Our data showed that Trdmt1 strongly responded in LPS-treated rats,and Trdmt1 knockout rats were vulnerable to LPS treat-ment with declined survival rate.We also observed more aggravated tissue damage and more cumulative functional cell degeneration in LPS-treated knockout rats compared with control rats.Further studies showed upregulated TNF-αlevel in liver,spleen,lung,and serum tissues,which may be explained by enhanced p65 and p38 phosphorylation.Conclusions:Our data demonstrated that Trdmt1 plays a protective role in inflamma-tion by regulating the TLR4-NF-κB/MAPK-TNF-αpathway.This work provides useful information to understand the TRDMT1 function in inflammation.展开更多
Coronary atherosclerotic disease is a serious disease in humans,but no suitable animal model is available currently for further studies.We used apolipoprotein E gene knockout（ApoE KO） rats to induce hypercholesterol...Coronary atherosclerotic disease is a serious disease in humans,but no suitable animal model is available currently for further studies.We used apolipoprotein E gene knockout（ApoE KO） rats to induce hypercholesterolemia through a special high cholesterol/bile salt diet（Paigen diet）,then analyzed aortic and coronaiy atherosclerosis lesions and the myocardial injury in order to establish a novel small animal model of coronary atherosclerosis.Plasma cholesterol of ApoE KO rats increased 7.6-fold compared with wild-type rats after 8 weeks on the Paigen diet.After 10 to 12 weeks of subsisting on the Paigen diet,ApoE KO rats developed mild aortic atherosclerosis with severe coronary atherosclerosis.Hematoxilyn and eosin staining showed that 11 out of 12 ApoE KO male rats had right coronary artery atherosclerosis,7 of them were〉70%occluded.Oil Red O（Lipid Stain）,Mac2 immuno-staining and Masson＇s tnchrome staining demonstrated substantial amounts of lipid,macrophages and collagen fibers in coronary atherosclerosis plaques.In addition,ApoE KO male rats had severe myocardial focal lesions with cholesterol ester as the main component in the lesions.In conclusion,ApoE KO rats developed severe hypercholesterolemia,coronary atherosclerosis and myocardial cholesterol ester deposition after subsisting on the Paigen diet and can be used as a novel animal model for studies on cholesterol metabolism and coronary atherosclerotic disease.展开更多
使用TALEN技术制作p53基因敲除大鼠模型,建立种群并分析大鼠表型。设计特异性识别p53基因外显子2的TALEN蛋白并构建相应载体,将体外转录的TALEN m RNA注射SD大鼠受精卵,出生后从仔鼠中通过测序筛选靶位点正确剪切的阳性小鼠。结果显示,...使用TALEN技术制作p53基因敲除大鼠模型,建立种群并分析大鼠表型。设计特异性识别p53基因外显子2的TALEN蛋白并构建相应载体,将体外转录的TALEN m RNA注射SD大鼠受精卵,出生后从仔鼠中通过测序筛选靶位点正确剪切的阳性小鼠。结果显示,注射得到11只新生仔鼠,通过测序发现其中有10只靶位点发生剪切。选取其中4只作为首建鼠进行繁殖。p53基因敲除纯合子表现出肿瘤易发性,主要自发性肿瘤类型为恶性纤维组织肉瘤。此外,p53基因敲除纯合子大鼠表现出眼睛发育异常,视网膜变性及晶状体纤维排列紊乱。通过TALEN技术高效地获得了p53基因敲除大鼠模型,纯合子敲除大鼠除了易发肿瘤并伴有眼发育异常,因而该敲除大鼠模型除了可用于研究肿瘤发生机制外,也可用于研究p53基因在眼发育过程中的功能。展开更多
目的观察搜风祛痰中药复方稳斑汤对ApoE基因敲除小鼠动脉粥样硬化(AS)不稳定斑块动物模型的影响。方法 6～8周龄ApoE基因敲除小鼠制备AS不稳定斑块模型,分别给予阿托伐他汀以及不同剂量稳斑汤干预1个月,正常组采用生理盐水灌胃1个月。...目的观察搜风祛痰中药复方稳斑汤对ApoE基因敲除小鼠动脉粥样硬化(AS)不稳定斑块动物模型的影响。方法 6～8周龄ApoE基因敲除小鼠制备AS不稳定斑块模型,分别给予阿托伐他汀以及不同剂量稳斑汤干预1个月,正常组采用生理盐水灌胃1个月。取小鼠主动脉组织HE染色进行病理学观察,并采用免疫组化和RT-PCR技术检测HO-1和TLR4的表达水平。结果各组小鼠腹主动脉组织中HO-1的表达水平治疗组明显高于对照组(P<0.01),各组小鼠腹主动脉组织中TLR4的表达水平治疗组明显低于对照组(P<0.01)。结论 Apo E基因敲除小鼠AS不稳定斑块的形成与炎症因子密切相关,搜风祛痰中药复方稳斑汤可能通过调节HO-1和TLR4的表达而干预AS斑块形成及破裂。展开更多
基金funded by the National Natural Science Foundation of China(31572378)the Major National Scientific Research Projects,China(2015CB943101)the Agricultural Science and Technology Innovation Program,China(ASTIP-IAS05)
文摘Patatin-like phospholipase domain containing 5 （PNPLA5） is a neotype neutral lipase with dual activity of anabolism and catabolism in vitro and in vivo, which has a low mRNA expression level in humans and mice. PNPLA5, which is localized to lipid droplets and required for efficient autophagy by optimal initiation, has been speculated to possess triglyceride hydro- lase activity, and has been associated with low density lipoprotein cholesterol （LDL-C）. Above all, PNPLA5 is a relatively new gene, which is reported less about its biological function research, especially the function research in the rats is still blank. In this study, we examined the spatiotemporal expression profile of PNPLA5 and found that it was expressed at low levels in most organs of Sprague Dawley （SD） rats, but was present at very high levels in the skin and testes. To furth.er determine the biological function of PNPLA5 in mammals, we generated PNPLA5-knockout SD rats using the clustered regularly-interspaced short palindromic repeats （CRISPR）/Cas9 system. PNPLA5-null rats were viable, but showed a variety of phenotypic abnormalities, such as abnormal bleeding, and varied hematobiochemical parameters including increased serum total cholesterol （TC）, tdglycerides and high density lipoprotein cholesterol （HDL-C） level, and reduced LDL-C level, compared with wild-type control rats. These data are consistent with an important role for PNPLA5 in lipid metabolism, provJdJng a new target gene and animal model for treatment of cardiovascular diseases in the future.
基金CAMS Innovation Fund for Medical Sciences(CIFMS)(2021-I 2M-1-024 and 2021-I 2M-1-034)and Beijing Municipal Natural Science Foundation(M21004)+1 种基金National Natural Science Foundation of China(31970508)111 Project of the Ministry of Education(B20095)。
文摘Background:Inflammation is a complex physiological and pathological process.Although many types of inflammation are well characterized,their physiological func-tions are largely unknown.tRNA aspartic acid methyltransferase 1(TRDMT1)has been implicated as a stress-related protein,but its intrinsic biological role is unclear.Methods:We constructed a Trdmt1 knockout rat and adopted the LPS-induced sepsis model.Survival curve,histopathological examination,expression of inflammatory fac-tors,and protein level of TLR4 pathway were analyzed.Results:Trdmt1 deletion had no obvious impact on development and growth.Trdmt1 de-letion slightly increased the mortality during aging.Our data showed that Trdmt1 strongly responded in LPS-treated rats,and Trdmt1 knockout rats were vulnerable to LPS treat-ment with declined survival rate.We also observed more aggravated tissue damage and more cumulative functional cell degeneration in LPS-treated knockout rats compared with control rats.Further studies showed upregulated TNF-αlevel in liver,spleen,lung,and serum tissues,which may be explained by enhanced p65 and p38 phosphorylation.Conclusions:Our data demonstrated that Trdmt1 plays a protective role in inflamma-tion by regulating the TLR4-NF-κB/MAPK-TNF-αpathway.This work provides useful information to understand the TRDMT1 function in inflammation.
文摘Coronary atherosclerotic disease is a serious disease in humans,but no suitable animal model is available currently for further studies.We used apolipoprotein E gene knockout（ApoE KO） rats to induce hypercholesterolemia through a special high cholesterol/bile salt diet（Paigen diet）,then analyzed aortic and coronaiy atherosclerosis lesions and the myocardial injury in order to establish a novel small animal model of coronary atherosclerosis.Plasma cholesterol of ApoE KO rats increased 7.6-fold compared with wild-type rats after 8 weeks on the Paigen diet.After 10 to 12 weeks of subsisting on the Paigen diet,ApoE KO rats developed mild aortic atherosclerosis with severe coronary atherosclerosis.Hematoxilyn and eosin staining showed that 11 out of 12 ApoE KO male rats had right coronary artery atherosclerosis,7 of them were〉70%occluded.Oil Red O（Lipid Stain）,Mac2 immuno-staining and Masson＇s tnchrome staining demonstrated substantial amounts of lipid,macrophages and collagen fibers in coronary atherosclerosis plaques.In addition,ApoE KO male rats had severe myocardial focal lesions with cholesterol ester as the main component in the lesions.In conclusion,ApoE KO rats developed severe hypercholesterolemia,coronary atherosclerosis and myocardial cholesterol ester deposition after subsisting on the Paigen diet and can be used as a novel animal model for studies on cholesterol metabolism and coronary atherosclerotic disease.
文摘使用TALEN技术制作p53基因敲除大鼠模型,建立种群并分析大鼠表型。设计特异性识别p53基因外显子2的TALEN蛋白并构建相应载体,将体外转录的TALEN m RNA注射SD大鼠受精卵,出生后从仔鼠中通过测序筛选靶位点正确剪切的阳性小鼠。结果显示,注射得到11只新生仔鼠,通过测序发现其中有10只靶位点发生剪切。选取其中4只作为首建鼠进行繁殖。p53基因敲除纯合子表现出肿瘤易发性,主要自发性肿瘤类型为恶性纤维组织肉瘤。此外,p53基因敲除纯合子大鼠表现出眼睛发育异常,视网膜变性及晶状体纤维排列紊乱。通过TALEN技术高效地获得了p53基因敲除大鼠模型,纯合子敲除大鼠除了易发肿瘤并伴有眼发育异常,因而该敲除大鼠模型除了可用于研究肿瘤发生机制外,也可用于研究p53基因在眼发育过程中的功能。
文摘目的观察搜风祛痰中药复方稳斑汤对ApoE基因敲除小鼠动脉粥样硬化(AS)不稳定斑块动物模型的影响。方法 6～8周龄ApoE基因敲除小鼠制备AS不稳定斑块模型,分别给予阿托伐他汀以及不同剂量稳斑汤干预1个月,正常组采用生理盐水灌胃1个月。取小鼠主动脉组织HE染色进行病理学观察,并采用免疫组化和RT-PCR技术检测HO-1和TLR4的表达水平。结果各组小鼠腹主动脉组织中HO-1的表达水平治疗组明显高于对照组(P<0.01),各组小鼠腹主动脉组织中TLR4的表达水平治疗组明显低于对照组(P<0.01)。结论 Apo E基因敲除小鼠AS不稳定斑块的形成与炎症因子密切相关,搜风祛痰中药复方稳斑汤可能通过调节HO-1和TLR4的表达而干预AS斑块形成及破裂。
