Eur Rev Med Pharmacol Sci:研究发现miR-374b可以通过降解PTEN促进MSCs的成骨分化

2018-06-22 MedSci MedSci原创

本研究旨在探究miR-374b是否通过介导PTEN促进MSCs分化为成骨细胞,从而促进骨折愈合。 提取小鼠间充质干细胞进行实验。流式细胞术确定MSCs表面抗原的表达以鉴定其纯度。使用茜素红染色来检测MSCs是否可以分化为成骨细胞。QRT-PCR检测骨折小鼠模型骨髓MSCs中成骨标志物基因的表达以及miR-374b和PTEN的表达。ALP活性检测试剂盒用于检测细胞中的ALP活性。 Western印迹

本研究旨在探究miR-374b是否通过介导PTEN促进MSCs分化为成骨细胞,从而促进骨折愈合。

提取小鼠间充质干细胞进行实验。流式细胞术确定MSCs表面抗原的表达以鉴定其纯度。使用茜素红染色来检测MSCs是否可以分化为成骨细胞。QRT-PCR检测骨折小鼠模型骨髓MSCs中成骨标志物基因的表达以及miR-374b和PTEN的表达。ALP活性检测试剂盒用于检测细胞中的ALP活性。 Western印迹评估细胞中成骨蛋白的变化。使用生物信息学方法来预测miR-374与靶基因之间的结合位点。使用萤光素酶报告基因测定法来确认miR-374是否可以与靶基因结合。

结果显示,在正常培养下,第4天MSCs长成长梭形。在成骨诱导培养基中诱导7天后,出现钙化结节。MSCs表面抗原标志物检测结果显示CD90为99.12%,CD45为0.23%。茜素红染色显示MSCs具有分化成成骨的能力。MiR-374b和PTEN的表达水平在小鼠骨折早期明显增加,但在后期没有观察到明显差异。过表达miR-374b后,细胞ALP活性、成骨相关基因和成骨相关蛋白的表达显著增加,而敲除miR-374b后,观察到相反的结果。萤光素酶报告测定的结果显示miR-374b可结合PTEN。如上所述,miR-374b的过表达可显著增加成骨相关基因和蛋白质的表达,而PTEN和miR-374b的过度表达可部分逆转结果。

综上所述,该研究结果表明,miR-374b可以通过降解PTEN促进MSCs的成骨分化,从而促进骨折愈合。

原始出处:


Ge JB, Lin JT, et al., MiR-374b promotes osteogenic differentiation of MSCs by degrading PTEN and promoting fracture healing
. Eur Rev Med Pharmacol Sci. 2018 Jun;22(11):3303-3310. doi: 10.26355/eurrev_201806_15149.

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    2019-06-05 HinsMax
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    2019-05-11 yb6560
  4. [GetPortalCommentsPageByObjectIdResponse(id=1821314, encodeId=6159182131454, content=<a href='/topic/show?id=428a101612e7' target=_blank style='color:#2F92EE;'>#骨分化#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=0, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=101612, encryptionId=428a101612e7, topicName=骨分化)], attachment=null, authenticateStatus=null, createdAvatar=http://thirdwx.qlogo.cn/mmopen/vi_32/qXbLjJB4FSzhQS3C5qQFMJkqicRIJFL6zHFwwhv0I95nzA6xicevQcEica0gk4A58UkbMMriadAgTY2b7N7Yd0ZsQg/132, createdBy=910c2500152, createdName=HinsMax, createdTime=Wed Jun 05 13:37:00 CST 2019, time=2019-06-05, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1788317, encodeId=d8821e883173b, content=<a href='/topic/show?id=38e014162e2' target=_blank style='color:#2F92EE;'>#Pharmacol#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=30, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14162, encryptionId=38e014162e2, topicName=Pharmacol)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=930e243, createdName=dingxiaobo, createdTime=Wed Oct 31 12:37:00 CST 2018, time=2018-10-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1996178, encodeId=b1d119961e88b, content=<a href='/topic/show?id=ed881120ec8' target=_blank style='color:#2F92EE;'>#MAC#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=22, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=11207, encryptionId=ed881120ec8, topicName=MAC)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4b65161, createdName=yb6560, createdTime=Sat May 11 08:37:00 CST 2019, time=2019-05-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1930627, encodeId=744c193062e68, content=<a href='/topic/show?id=22f61180211' target=_blank style='color:#2F92EE;'>#miR#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=21, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=11802, encryptionId=22f61180211, topicName=miR)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=0a27288, createdName=smallant2002, createdTime=Thu Apr 18 10:37:00 CST 2019, time=2019-04-18, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1908437, encodeId=bd45190843e07, content=<a href='/topic/show?id=a8d0141605e' target=_blank style='color:#2F92EE;'>#pharma#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=30, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14160, encryptionId=a8d0141605e, topicName=pharma)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=f634250, createdName=chentianping, createdTime=Sat Aug 18 04:37:00 CST 2018, time=2018-08-18, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1753411, encodeId=e4ee1e5341116, content=<a href='/topic/show?id=6a57e33772b' target=_blank style='color:#2F92EE;'>#研究发现#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=16, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=73377, encryptionId=6a57e33772b, topicName=研究发现)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=7e1736602208, createdName=hanhaisha2020, createdTime=Mon Apr 08 01:37:00 CST 2019, time=2019-04-08, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1787489, encodeId=bb6c1e8748956, content=<a href='/topic/show?id=fa03141591e' target=_blank style='color:#2F92EE;'>#Pharm#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14159, encryptionId=fa03141591e, topicName=Pharm)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=8942242, createdName=jj000001, createdTime=Sun Apr 21 10:37:00 CST 2019, time=2019-04-21, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1315924, encodeId=5a31131592478, content=<a href='/topic/show?id=134f14834f2' target=_blank style='color:#2F92EE;'>#PTEN#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=31, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14834, encryptionId=134f14834f2, topicName=PTEN)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=3f67438, createdName=xiaogang317, createdTime=Sun Jun 24 07:37:00 CST 2018, time=2018-06-24, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1337459, encodeId=c589133e4599b, content=<a href='/topic/show?id=863f5409610' target=_blank style='color:#2F92EE;'>#成骨分化#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=54096, encryptionId=863f5409610, topicName=成骨分化)], attachment=null, authenticateStatus=null, createdAvatar=http://thirdwx.qlogo.cn/mmopen/vi_32/DYAIOgq83eqySEUYKeYDjXJMYMrtscBNkpAnLfQZgDcdkEratcJPqS7ARJOXqllZQhCSFicErlBFbIywuoiaP0LQ/132, createdBy=012d2500132, createdName=ms5880526374914242, createdTime=Sun Jun 24 07:37:00 CST 2018, time=2018-06-24, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1351736, encodeId=a3671351e3698, content=<a href='/topic/show?id=e95954093cd' target=_blank style='color:#2F92EE;'>#成骨#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=54093, encryptionId=e95954093cd, topicName=成骨)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=zhaojie88, createdTime=Sun Jun 24 07:37:00 CST 2018, time=2018-06-24, status=1, ipAttribution=)]
    2019-04-18 smallant2002
  5. 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encodeId=e4ee1e5341116, content=<a href='/topic/show?id=6a57e33772b' target=_blank style='color:#2F92EE;'>#研究发现#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=16, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=73377, encryptionId=6a57e33772b, topicName=研究发现)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=7e1736602208, createdName=hanhaisha2020, createdTime=Mon Apr 08 01:37:00 CST 2019, time=2019-04-08, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1787489, encodeId=bb6c1e8748956, content=<a href='/topic/show?id=fa03141591e' target=_blank style='color:#2F92EE;'>#Pharm#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14159, encryptionId=fa03141591e, topicName=Pharm)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=8942242, createdName=jj000001, createdTime=Sun Apr 21 10:37:00 CST 2019, time=2019-04-21, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1315924, encodeId=5a31131592478, content=<a href='/topic/show?id=134f14834f2' target=_blank style='color:#2F92EE;'>#PTEN#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=31, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14834, encryptionId=134f14834f2, topicName=PTEN)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=3f67438, createdName=xiaogang317, createdTime=Sun Jun 24 07:37:00 CST 2018, time=2018-06-24, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1337459, encodeId=c589133e4599b, content=<a href='/topic/show?id=863f5409610' target=_blank style='color:#2F92EE;'>#成骨分化#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=54096, encryptionId=863f5409610, topicName=成骨分化)], attachment=null, authenticateStatus=null, createdAvatar=http://thirdwx.qlogo.cn/mmopen/vi_32/DYAIOgq83eqySEUYKeYDjXJMYMrtscBNkpAnLfQZgDcdkEratcJPqS7ARJOXqllZQhCSFicErlBFbIywuoiaP0LQ/132, createdBy=012d2500132, createdName=ms5880526374914242, createdTime=Sun Jun 24 07:37:00 CST 2018, time=2018-06-24, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1351736, encodeId=a3671351e3698, content=<a href='/topic/show?id=e95954093cd' target=_blank style='color:#2F92EE;'>#成骨#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=54093, encryptionId=e95954093cd, topicName=成骨)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=zhaojie88, createdTime=Sun Jun 24 07:37:00 CST 2018, time=2018-06-24, status=1, ipAttribution=)]
  6. [GetPortalCommentsPageByObjectIdResponse(id=1821314, encodeId=6159182131454, content=<a href='/topic/show?id=428a101612e7' target=_blank style='color:#2F92EE;'>#骨分化#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=0, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=101612, encryptionId=428a101612e7, topicName=骨分化)], attachment=null, authenticateStatus=null, createdAvatar=http://thirdwx.qlogo.cn/mmopen/vi_32/qXbLjJB4FSzhQS3C5qQFMJkqicRIJFL6zHFwwhv0I95nzA6xicevQcEica0gk4A58UkbMMriadAgTY2b7N7Yd0ZsQg/132, createdBy=910c2500152, createdName=HinsMax, createdTime=Wed Jun 05 13:37:00 CST 2019, time=2019-06-05, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1788317, encodeId=d8821e883173b, content=<a href='/topic/show?id=38e014162e2' target=_blank style='color:#2F92EE;'>#Pharmacol#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=30, replyNumber=0, topicName=null, topicId=null, 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  7. [GetPortalCommentsPageByObjectIdResponse(id=1821314, encodeId=6159182131454, content=<a href='/topic/show?id=428a101612e7' target=_blank style='color:#2F92EE;'>#骨分化#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=0, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=101612, encryptionId=428a101612e7, topicName=骨分化)], attachment=null, authenticateStatus=null, createdAvatar=http://thirdwx.qlogo.cn/mmopen/vi_32/qXbLjJB4FSzhQS3C5qQFMJkqicRIJFL6zHFwwhv0I95nzA6xicevQcEica0gk4A58UkbMMriadAgTY2b7N7Yd0ZsQg/132, createdBy=910c2500152, createdName=HinsMax, createdTime=Wed Jun 05 13:37:00 CST 2019, time=2019-06-05, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1788317, encodeId=d8821e883173b, content=<a href='/topic/show?id=38e014162e2' target=_blank style='color:#2F92EE;'>#Pharmacol#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=30, replyNumber=0, topicName=null, topicId=null, 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21 10:37:00 CST 2019, time=2019-04-21, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1315924, encodeId=5a31131592478, content=<a href='/topic/show?id=134f14834f2' target=_blank style='color:#2F92EE;'>#PTEN#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=31, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14834, encryptionId=134f14834f2, topicName=PTEN)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=3f67438, createdName=xiaogang317, createdTime=Sun Jun 24 07:37:00 CST 2018, time=2018-06-24, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1337459, encodeId=c589133e4599b, content=<a href='/topic/show?id=863f5409610' target=_blank style='color:#2F92EE;'>#成骨分化#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=54096, encryptionId=863f5409610, topicName=成骨分化)], attachment=null, authenticateStatus=null, createdAvatar=http://thirdwx.qlogo.cn/mmopen/vi_32/DYAIOgq83eqySEUYKeYDjXJMYMrtscBNkpAnLfQZgDcdkEratcJPqS7ARJOXqllZQhCSFicErlBFbIywuoiaP0LQ/132, createdBy=012d2500132, createdName=ms5880526374914242, createdTime=Sun Jun 24 07:37:00 CST 2018, time=2018-06-24, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1351736, encodeId=a3671351e3698, content=<a href='/topic/show?id=e95954093cd' target=_blank style='color:#2F92EE;'>#成骨#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=54093, encryptionId=e95954093cd, topicName=成骨)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=zhaojie88, createdTime=Sun Jun 24 07:37:00 CST 2018, time=2018-06-24, status=1, ipAttribution=)]
    2019-04-21 jj000001
  8. 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21 10:37:00 CST 2019, time=2019-04-21, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1315924, encodeId=5a31131592478, content=<a href='/topic/show?id=134f14834f2' target=_blank style='color:#2F92EE;'>#PTEN#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=31, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14834, encryptionId=134f14834f2, topicName=PTEN)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=3f67438, createdName=xiaogang317, createdTime=Sun Jun 24 07:37:00 CST 2018, time=2018-06-24, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1337459, encodeId=c589133e4599b, content=<a href='/topic/show?id=863f5409610' target=_blank style='color:#2F92EE;'>#成骨分化#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=54096, encryptionId=863f5409610, topicName=成骨分化)], attachment=null, authenticateStatus=null, createdAvatar=http://thirdwx.qlogo.cn/mmopen/vi_32/DYAIOgq83eqySEUYKeYDjXJMYMrtscBNkpAnLfQZgDcdkEratcJPqS7ARJOXqllZQhCSFicErlBFbIywuoiaP0LQ/132, createdBy=012d2500132, createdName=ms5880526374914242, createdTime=Sun Jun 24 07:37:00 CST 2018, time=2018-06-24, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1351736, encodeId=a3671351e3698, content=<a href='/topic/show?id=e95954093cd' target=_blank style='color:#2F92EE;'>#成骨#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=54093, encryptionId=e95954093cd, topicName=成骨)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=zhaojie88, createdTime=Sun Jun 24 07:37:00 CST 2018, time=2018-06-24, status=1, ipAttribution=)]
    2018-06-24 xiaogang317
  9. 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  10. 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    2018-06-24 zhaojie88