目的:应用肌性斜颈兔动物模型,探讨针刀治疗肌性斜颈的作用机制及意义。方法:无水酒精注射制备动物肌性斜颈兔子模型后,选取模型兔子48只,随机分为正常组、模型组、针刀组、按摩组,每组12只。分别给予各组对应治疗,治疗2个月后各组分别...目的:应用肌性斜颈兔动物模型,探讨针刀治疗肌性斜颈的作用机制及意义。方法:无水酒精注射制备动物肌性斜颈兔子模型后,选取模型兔子48只,随机分为正常组、模型组、针刀组、按摩组,每组12只。分别给予各组对应治疗,治疗2个月后各组分别取部分胸锁乳突肌制作标本,免疫组化法观察钙蛋白酶1(calpain-1)、分化抗原决定簇45(CD45)、基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)、基质金属蛋白酶抑制因子2(tissue inhibitor of metalloproteinases-2,TIMP-2)在各组肌纤维中的表达情况。结果:与正常组比较,其余各组实验动物明显出现颈部活动受限、斜颈、头颈不对称症状,而治疗结束后,针刀组和按摩组实验动物颈部活动受限改善,无明显头颈不对称症状。calpain-1、CD45、TIMP-2的表达,与正常组比较,模型组、按摩组上调(P<0.01);与模型组比较,针刀组、按摩组下调(P<0.01);与针刀组比较,按摩组上调(P<0.01)。MMP-2表达,与正常组比较,模型组下调(P<0.01),与模型组比较,针刀组、按摩组上调(P<0.05)。结论:针刀治疗肌性斜颈效果显著,其作用机制可能与减少calpain-1、CD45、TIMP-2表达,增加MMP-2表达有关。展开更多
The interaction between myocytes and intramuscular adipocytes is a hot scientific topic. Using a co-culture system, this study aims to investigate the regulation of intramuscular fat deposition in chicken muscle tissu...The interaction between myocytes and intramuscular adipocytes is a hot scientific topic. Using a co-culture system, this study aims to investigate the regulation of intramuscular fat deposition in chicken muscle tissue through the interaction between myocyte and adipocyte and identify important intermediary regulatory factors. Our proteomics data showed that the protein expression of tissue inhibitor of metalloproteinases 2(TIMP2) increased significantly in the culture medium of the co-culture system, and the content of lipid droplets was more in the co-culture intramuscular adipocytes. In addition, TIMP2 was significantly upregulated(P<0.01) in muscle tissue of individuals with high intramuscular fat content.Weighted gene co-expression network analysis revealed that TIMP2 was mainly involved in the extracellular matrix receptor interaction signaling pathway and its expression was significantly correlated with triglyceride, intramuscular fat,C14:0, C14:1, C16:0, C16:1, and C18:1n9C levels. Additionally, TIMP2 was co-expressed with various representative genes related to lipid metabolism(such as ADIPOQ, SCD, ELOVL5, ELOVL7, and LPL), as well as certain genes involved in extracellular matrix receptor interaction(such as COL1A2, COL4A2, COL5A1, COL6A1, and COL6A3), which are also significantly upregulated(P<0.05 or P<0.01) in muscle tissue of individuals with high intramuscular fat content.Our findings reveal that TIMP2 promotes intramuscular fat deposition in muscle tissue through the extracellular matrix receptor interaction signaling pathway.展开更多
Objective:To investigate whether miR-483-5p regulates osteoclast generation by targeting Timp2.miR-483-5p can promote osteoclast differentiation and bone destruction.Methods:Target genes of miR-483-5p were predicted b...Objective:To investigate whether miR-483-5p regulates osteoclast generation by targeting Timp2.miR-483-5p can promote osteoclast differentiation and bone destruction.Methods:Target genes of miR-483-5p were predicted by miRNAs target gene prediction software TargetScan8.0,and wild type and mutant 3'UTR plasmids were constructed.Dual luciferase reporter genes were used to verify whether target genes had a targeted regulatory relationship with miR-483-5p.Western blotting was used to detect the corresponding changes in the expression level of target protein after adjusting the level of miR-483-5p in cells.Cells were transfected or infected with target gene siRNA or target protein lentivirus,and TRAP staining and q-PCR assays were performed.In addition,for osteoclast induction experiment,RAW264.7 cells were co-transfected with ago-miR-483-5p and target protein-overexpressed lentiviruses q-PCR and TRAP staining were performed respectively.Results:Bioinformatics software was used to predict the target gene of miR-483-5p,and the Timp2 gene was found to regulate osteoclasts,and the dual luciferase reporter detection system found that miR-483-5p could be associated with the 3-UTR of the predicted target gene Timp2 gene.There are complementary loci and targeted regulatory relationship between them.Subsequently,we upregulated miR-483-5p in RAW264.7 cells to reduce the expression of Timp2.Compared with the normal group,the number of osteoclasts and the expression of osteoclast-specific genes increased significantly after the induction of Timp2 in knockdown cells.After co-transfection of target gene and miR-483-5p into cells,the number of osteoclasts and the expression of specific genes decreased significantly compared with the normal group.Conclusion:Timp2 is a downstream target gene of miR-483-5p and is involved in and inhibits osteoclast generation.展开更多
文摘目的:应用肌性斜颈兔动物模型,探讨针刀治疗肌性斜颈的作用机制及意义。方法:无水酒精注射制备动物肌性斜颈兔子模型后,选取模型兔子48只,随机分为正常组、模型组、针刀组、按摩组,每组12只。分别给予各组对应治疗,治疗2个月后各组分别取部分胸锁乳突肌制作标本,免疫组化法观察钙蛋白酶1(calpain-1)、分化抗原决定簇45(CD45)、基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)、基质金属蛋白酶抑制因子2(tissue inhibitor of metalloproteinases-2,TIMP-2)在各组肌纤维中的表达情况。结果:与正常组比较,其余各组实验动物明显出现颈部活动受限、斜颈、头颈不对称症状,而治疗结束后,针刀组和按摩组实验动物颈部活动受限改善,无明显头颈不对称症状。calpain-1、CD45、TIMP-2的表达,与正常组比较,模型组、按摩组上调(P<0.01);与模型组比较,针刀组、按摩组下调(P<0.01);与针刀组比较,按摩组上调(P<0.01)。MMP-2表达,与正常组比较,模型组下调(P<0.01),与模型组比较,针刀组、按摩组上调(P<0.05)。结论:针刀治疗肌性斜颈效果显著,其作用机制可能与减少calpain-1、CD45、TIMP-2表达,增加MMP-2表达有关。
基金funded by the grants from the National Natural Science Foundation of China(31872340)the State Key Laboratory of Animal Nutrition(2004DA125184G2109)+1 种基金the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences(ASTIP-IAS04)the earmarked fund for China Agriculture Research System(CARS-41)。
文摘The interaction between myocytes and intramuscular adipocytes is a hot scientific topic. Using a co-culture system, this study aims to investigate the regulation of intramuscular fat deposition in chicken muscle tissue through the interaction between myocyte and adipocyte and identify important intermediary regulatory factors. Our proteomics data showed that the protein expression of tissue inhibitor of metalloproteinases 2(TIMP2) increased significantly in the culture medium of the co-culture system, and the content of lipid droplets was more in the co-culture intramuscular adipocytes. In addition, TIMP2 was significantly upregulated(P<0.01) in muscle tissue of individuals with high intramuscular fat content.Weighted gene co-expression network analysis revealed that TIMP2 was mainly involved in the extracellular matrix receptor interaction signaling pathway and its expression was significantly correlated with triglyceride, intramuscular fat,C14:0, C14:1, C16:0, C16:1, and C18:1n9C levels. Additionally, TIMP2 was co-expressed with various representative genes related to lipid metabolism(such as ADIPOQ, SCD, ELOVL5, ELOVL7, and LPL), as well as certain genes involved in extracellular matrix receptor interaction(such as COL1A2, COL4A2, COL5A1, COL6A1, and COL6A3), which are also significantly upregulated(P<0.05 or P<0.01) in muscle tissue of individuals with high intramuscular fat content.Our findings reveal that TIMP2 promotes intramuscular fat deposition in muscle tissue through the extracellular matrix receptor interaction signaling pathway.
基金National Natural Science Foundation of China(No.81860645)Hainan Medical University Introduced Talents Research Start-Up Funds(No.2015)。
文摘Objective:To investigate whether miR-483-5p regulates osteoclast generation by targeting Timp2.miR-483-5p can promote osteoclast differentiation and bone destruction.Methods:Target genes of miR-483-5p were predicted by miRNAs target gene prediction software TargetScan8.0,and wild type and mutant 3'UTR plasmids were constructed.Dual luciferase reporter genes were used to verify whether target genes had a targeted regulatory relationship with miR-483-5p.Western blotting was used to detect the corresponding changes in the expression level of target protein after adjusting the level of miR-483-5p in cells.Cells were transfected or infected with target gene siRNA or target protein lentivirus,and TRAP staining and q-PCR assays were performed.In addition,for osteoclast induction experiment,RAW264.7 cells were co-transfected with ago-miR-483-5p and target protein-overexpressed lentiviruses q-PCR and TRAP staining were performed respectively.Results:Bioinformatics software was used to predict the target gene of miR-483-5p,and the Timp2 gene was found to regulate osteoclasts,and the dual luciferase reporter detection system found that miR-483-5p could be associated with the 3-UTR of the predicted target gene Timp2 gene.There are complementary loci and targeted regulatory relationship between them.Subsequently,we upregulated miR-483-5p in RAW264.7 cells to reduce the expression of Timp2.Compared with the normal group,the number of osteoclasts and the expression of osteoclast-specific genes increased significantly after the induction of Timp2 in knockdown cells.After co-transfection of target gene and miR-483-5p into cells,the number of osteoclasts and the expression of specific genes decreased significantly compared with the normal group.Conclusion:Timp2 is a downstream target gene of miR-483-5p and is involved in and inhibits osteoclast generation.
