The previous study of Smad3 gene knockout mice ( Smad3 ex8/ex8 )shows that the Smad3 ex8/ex8 mice develop progressive leukocytosis, periodontitis, gastritis, colitis and chronic infection with abscess formation adjace...The previous study of Smad3 gene knockout mice ( Smad3 ex8/ex8 )shows that the Smad3 ex8/ex8 mice develop progressive leukocytosis, periodontitis, gastritis, colitis and chronic infection with abscess formation adjacent to mucosal surfaces .symptomatic mutant mice exhibit thymic involution, enlarged lymph nodes and T cells with activated phenotype. Further study suggests that the thymic cells and peripheral T ceels of Smad3 ex8/ex8 mice have lost the response to TGF β.Furthermore, nwe found that the homologous Smad3 ex8/ex mice developed degenerative joint disease resembling human osteoarthritis, osteoporosis and wound healing up quicker. So the mice can serve as an ideal animal model for immune dysregulation, osteoarthritis and so on. To futher study the important role of Smad3 during the vertebrate development ,we charactered the genotype and reproduce the Smad3 knockout mice .Compare to the general experimental mice, the mice of genetic modifications is different in the breeding and reproduction. Besider the reproduction of inbred lines, we also must keep the mice inherit the mutant gene and consider the influence of mutant gene to the mouse’s reproduction. Using PCR and Southern blot to characterize the genotype of the offspring can solve the question .Just like the homozygote of Smad3 ex8/ex8 mice lacking fertility, the homozygous mice cann’t be used for propagating. We use the heterozygous mice for propagating, while the homozygous and wild type mice were used for phenotype analysis. The Smad3 ex8/ex8 heterzyous mice were used for breeding. With genotype being characterized, we found that the ratio of 3 genotype of the Smad3 ex8/ex8 heterzyous mice’ offsprings fits the Mendel’s laws. Their gestation is the sames as others mice,and embryo interval and litter are not different from others, We kept means of cryopreservation by vitrification of embryos and got normal conditions .Using the Smad3 ex8/ex8 heterzyous mice for propagating, we can be used for keeping breed and propagating.展开更多
Previous studies have shown that up-regulation of transforming growth factor β1 results in neuroprotective effects. However, the role of the transforming growth factor β1 downstream molecule, SMAD2/3, following isch...Previous studies have shown that up-regulation of transforming growth factor β1 results in neuroprotective effects. However, the role of the transforming growth factor β1 downstream molecule, SMAD2/3, following ischemia/reperfusion remains unclear. Here, we investigated the neuroprotective effects of SMAD2/3 by analyzing the relationships between SMAD2/3 expression and cell apoptosis and inflammation in the brain of a rat model of cerebral ischemia/reperfusion. Levels of SMAD2/3 m RNA were up-regulated in the ischemic penumbra 6 hours after cerebral ischemia/reperfusion, reached a peak after 72 hours and were then decreased at 7 days. Phosphorylated SMAD2/3 protein levels at the aforementioned time points were consistent with the m RNA levels. Over-expression of SMAD3 in the brains of the ischemia/reperfusion model rats via delivery of an adeno-associated virus containing the SMAD3 gene could reduce tumor necrosis factor-α and interleukin-1β m RNA levels, down-regulate expression of the pro-apoptotic gene, capase-3, and up-regulate expression of the anti-apoptotic protein, Bcl-2. The SMAD3 protein level was negatively correlated with cell apoptosis. These findings indicate that SMAD3 exhibits neuroprotective effects on the brain after ischemia/reperfusion through anti-inflammatory and anti-apoptotic pathways.展开更多
文摘The previous study of Smad3 gene knockout mice ( Smad3 ex8/ex8 )shows that the Smad3 ex8/ex8 mice develop progressive leukocytosis, periodontitis, gastritis, colitis and chronic infection with abscess formation adjacent to mucosal surfaces .symptomatic mutant mice exhibit thymic involution, enlarged lymph nodes and T cells with activated phenotype. Further study suggests that the thymic cells and peripheral T ceels of Smad3 ex8/ex8 mice have lost the response to TGF β.Furthermore, nwe found that the homologous Smad3 ex8/ex mice developed degenerative joint disease resembling human osteoarthritis, osteoporosis and wound healing up quicker. So the mice can serve as an ideal animal model for immune dysregulation, osteoarthritis and so on. To futher study the important role of Smad3 during the vertebrate development ,we charactered the genotype and reproduce the Smad3 knockout mice .Compare to the general experimental mice, the mice of genetic modifications is different in the breeding and reproduction. Besider the reproduction of inbred lines, we also must keep the mice inherit the mutant gene and consider the influence of mutant gene to the mouse’s reproduction. Using PCR and Southern blot to characterize the genotype of the offspring can solve the question .Just like the homozygote of Smad3 ex8/ex8 mice lacking fertility, the homozygous mice cann’t be used for propagating. We use the heterozygous mice for propagating, while the homozygous and wild type mice were used for phenotype analysis. The Smad3 ex8/ex8 heterzyous mice were used for breeding. With genotype being characterized, we found that the ratio of 3 genotype of the Smad3 ex8/ex8 heterzyous mice’ offsprings fits the Mendel’s laws. Their gestation is the sames as others mice,and embryo interval and litter are not different from others, We kept means of cryopreservation by vitrification of embryos and got normal conditions .Using the Smad3 ex8/ex8 heterzyous mice for propagating, we can be used for keeping breed and propagating.
基金supported by the National Natural Science Foundation of China,No.81460193
文摘Previous studies have shown that up-regulation of transforming growth factor β1 results in neuroprotective effects. However, the role of the transforming growth factor β1 downstream molecule, SMAD2/3, following ischemia/reperfusion remains unclear. Here, we investigated the neuroprotective effects of SMAD2/3 by analyzing the relationships between SMAD2/3 expression and cell apoptosis and inflammation in the brain of a rat model of cerebral ischemia/reperfusion. Levels of SMAD2/3 m RNA were up-regulated in the ischemic penumbra 6 hours after cerebral ischemia/reperfusion, reached a peak after 72 hours and were then decreased at 7 days. Phosphorylated SMAD2/3 protein levels at the aforementioned time points were consistent with the m RNA levels. Over-expression of SMAD3 in the brains of the ischemia/reperfusion model rats via delivery of an adeno-associated virus containing the SMAD3 gene could reduce tumor necrosis factor-α and interleukin-1β m RNA levels, down-regulate expression of the pro-apoptotic gene, capase-3, and up-regulate expression of the anti-apoptotic protein, Bcl-2. The SMAD3 protein level was negatively correlated with cell apoptosis. These findings indicate that SMAD3 exhibits neuroprotective effects on the brain after ischemia/reperfusion through anti-inflammatory and anti-apoptotic pathways.
文摘【目的】探讨补肾活血化痰法改善自发性高血压大鼠(SHR)左心室纤维化机制。【方法】选用20只自发性高血压大鼠,随机分为模型组和中药组,10只WKY大鼠(Wistar-Kyoto rat)作为正常对照组。干预12周后,应用Masson染色法检测大鼠左心室纤维化程度,逆转录—聚合酶链反应(RT-PCR)法检测大鼠左心室Smad3 m RNA表达水平,Western blot法检测大鼠左心室Smad3表达水平。【结果】中药组大鼠心肌纤维化水平、Smad3蛋白表达水平、Smad3基因表达水平显著低于模型组(均P<0.05)。【结论】补肾活血化痰法可能通过抑制Smad3表达改善自发性高血压大鼠左心室纤维化进程。
