目的研究ERK1/2信号通路在左归丸含药血清调控MC3T3-E1细胞基因表达中的作用。方法以倍美力为阳性对照药,对SD♀大鼠灌服高、中、低剂量的左归丸混悬液,d7腹主动脉取血分离含药血清。采用MTT法检测左归丸含药血清和PD98059对MC3T3-E1细...目的研究ERK1/2信号通路在左归丸含药血清调控MC3T3-E1细胞基因表达中的作用。方法以倍美力为阳性对照药,对SD♀大鼠灌服高、中、低剂量的左归丸混悬液,d7腹主动脉取血分离含药血清。采用MTT法检测左归丸含药血清和PD98059对MC3T3-E1细胞增殖作用的影响,采用Western blot法检测ERK1/2蛋白的磷酸化水平及TGF-β1、Smad4蛋白表达,采用Real Time RT-PCR法检测Cbfα1、COLⅠmRNA表达。结果左归丸含药血清和倍美力能促进MC3T3-E1细胞增殖,诱导ERK1/2蛋白的磷酸化,促进TGF-β1、Smad4蛋白分泌,上调其Cbfα1、COLⅠmRNA表达;加入PD98059后MC3T3-E1细胞增殖下降、ERK1/2蛋白的磷酸化水平降低、TGF-β1蛋白表达进一步升高、Smad4蛋白表达降低、Cbfα1、COLⅠmRNA表达下调。结论左归丸能有效促进成骨细胞增殖和分化;左归丸可能是通过发挥雌激素样作用调控了ERK/Smads信号通路,从而达到防治骨质疏松的目的。展开更多
目的研究补骨脂素对人皮肤成纤维细胞(ESF-1)衰老相关基因的影响。方法以雌二醇为阳性对照,采用MTT法检测补骨脂素高、中、低剂量组(100,10,1μmol·L-1)对细胞增殖的影响,用RT-PCR技术检测补骨脂素作用于ESF-1细胞后对衰老相关基...目的研究补骨脂素对人皮肤成纤维细胞(ESF-1)衰老相关基因的影响。方法以雌二醇为阳性对照,采用MTT法检测补骨脂素高、中、低剂量组(100,10,1μmol·L-1)对细胞增殖的影响,用RT-PCR技术检测补骨脂素作用于ESF-1细胞后对衰老相关基因Ⅰ型胶原蛋白(ColⅠ)、Ⅲ型胶原蛋白(ColⅢ)、基质金属蛋白酶1(MMP-1)、基质金属蛋白酶抑制剂1(TIMP-1)及基质金属蛋白酶抑制剂2(TIMP-2)mRNA表达水平的影响。结果补骨脂素中剂量组的作用与雌二醇相似,能使ESF-1细胞的增殖明显提高,同时使ColⅠ、ColⅢ、T I M P-1、TIMP-2 mRNA的表达水平显著增强,MMP-1 mRNA的表达水平显著降低。结论补骨脂素具有促E S F-1细胞增殖的作用,可促进胶原蛋白合成,提示其具有潜在的抗皮肤衰老作用。展开更多
Objective:To evaluate the effect of human umbilical cord mesenchymal stem cells(hUC-MSCs)on preventing rats from glucocorticoid-induced osteonecrosis of femoral head(GC-ONFH)in the early stage in vivo and to investiga...Objective:To evaluate the effect of human umbilical cord mesenchymal stem cells(hUC-MSCs)on preventing rats from glucocorticoid-induced osteonecrosis of femoral head(GC-ONFH)in the early stage in vivo and to investigate the possible mechanism of hUC-MSCs in regulating the balance of osteogenesis and adipogenesis.Methods:All rats were randomly divided into 3 groups:control group(C group),model group(M group),and intervention group(Ⅰ group).The model of GC-ONFH was developed by a sequential administration of lipopolysaccharide and methylprednisolone.The rats in the Ⅰ group were treated with caudal vein injection of hUC-MSCs.Six weeks later,the blood samples were obtained to measure the activity of alkaline phosphatase(ALP)and the content of triglyceride(TG)in serum,and the femoral heads were harvested and observed by hematoxylin-eosin staining,Micro-CT,Western blot and real-time quantitative polymerase chain reaction.Results:After intervention of hUC-MSCs,the necrosis rate of femoral head decreased from 83%(10/12)to 33%(4/12),the rate of empty bone lacuna was significantly decreased,the activity of ALP increased significantly,the content of TG decreased significantly,the bone density increased obviously,the expression of RUNX2 and Col Ⅰ increased significantly and the expression of PPARγ decreased significantly.Conclusion:These results revealed that caudal vein injection of hUC-MSCs can effectively reduce the incidence of GC-ONFH in rats by increasing ALP activity and reducing TG content in serum,increasing bone mineral density,promoting the expression of RUNX2 and Col I,and inhibiting the expression of PPARγ.展开更多
文摘目的研究ERK1/2信号通路在左归丸含药血清调控MC3T3-E1细胞基因表达中的作用。方法以倍美力为阳性对照药,对SD♀大鼠灌服高、中、低剂量的左归丸混悬液,d7腹主动脉取血分离含药血清。采用MTT法检测左归丸含药血清和PD98059对MC3T3-E1细胞增殖作用的影响,采用Western blot法检测ERK1/2蛋白的磷酸化水平及TGF-β1、Smad4蛋白表达,采用Real Time RT-PCR法检测Cbfα1、COLⅠmRNA表达。结果左归丸含药血清和倍美力能促进MC3T3-E1细胞增殖,诱导ERK1/2蛋白的磷酸化,促进TGF-β1、Smad4蛋白分泌,上调其Cbfα1、COLⅠmRNA表达;加入PD98059后MC3T3-E1细胞增殖下降、ERK1/2蛋白的磷酸化水平降低、TGF-β1蛋白表达进一步升高、Smad4蛋白表达降低、Cbfα1、COLⅠmRNA表达下调。结论左归丸能有效促进成骨细胞增殖和分化;左归丸可能是通过发挥雌激素样作用调控了ERK/Smads信号通路,从而达到防治骨质疏松的目的。
文摘目的研究补骨脂素对人皮肤成纤维细胞(ESF-1)衰老相关基因的影响。方法以雌二醇为阳性对照,采用MTT法检测补骨脂素高、中、低剂量组(100,10,1μmol·L-1)对细胞增殖的影响,用RT-PCR技术检测补骨脂素作用于ESF-1细胞后对衰老相关基因Ⅰ型胶原蛋白(ColⅠ)、Ⅲ型胶原蛋白(ColⅢ)、基质金属蛋白酶1(MMP-1)、基质金属蛋白酶抑制剂1(TIMP-1)及基质金属蛋白酶抑制剂2(TIMP-2)mRNA表达水平的影响。结果补骨脂素中剂量组的作用与雌二醇相似,能使ESF-1细胞的增殖明显提高,同时使ColⅠ、ColⅢ、T I M P-1、TIMP-2 mRNA的表达水平显著增强,MMP-1 mRNA的表达水平显著降低。结论补骨脂素具有促E S F-1细胞增殖的作用,可促进胶原蛋白合成,提示其具有潜在的抗皮肤衰老作用。
基金supported by the National Natural Science Foundation of China(No.81672154).
文摘Objective:To evaluate the effect of human umbilical cord mesenchymal stem cells(hUC-MSCs)on preventing rats from glucocorticoid-induced osteonecrosis of femoral head(GC-ONFH)in the early stage in vivo and to investigate the possible mechanism of hUC-MSCs in regulating the balance of osteogenesis and adipogenesis.Methods:All rats were randomly divided into 3 groups:control group(C group),model group(M group),and intervention group(Ⅰ group).The model of GC-ONFH was developed by a sequential administration of lipopolysaccharide and methylprednisolone.The rats in the Ⅰ group were treated with caudal vein injection of hUC-MSCs.Six weeks later,the blood samples were obtained to measure the activity of alkaline phosphatase(ALP)and the content of triglyceride(TG)in serum,and the femoral heads were harvested and observed by hematoxylin-eosin staining,Micro-CT,Western blot and real-time quantitative polymerase chain reaction.Results:After intervention of hUC-MSCs,the necrosis rate of femoral head decreased from 83%(10/12)to 33%(4/12),the rate of empty bone lacuna was significantly decreased,the activity of ALP increased significantly,the content of TG decreased significantly,the bone density increased obviously,the expression of RUNX2 and Col Ⅰ increased significantly and the expression of PPARγ decreased significantly.Conclusion:These results revealed that caudal vein injection of hUC-MSCs can effectively reduce the incidence of GC-ONFH in rats by increasing ALP activity and reducing TG content in serum,increasing bone mineral density,promoting the expression of RUNX2 and Col I,and inhibiting the expression of PPARγ.
