双氢睾酮体外培养人毛囊中差异表达的miR-133b对人毛乳头细胞的增殖及诱导能力的影响

邓文佳; 邓长健; 韩乐; 刘本; 唐鑫; 万苗坚

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双氢睾酮体外培养人毛囊中差异表达的miR-133b对人毛乳头细胞的增殖及诱导能力的影响

基础研究 | 更新时间：2023-11-01

- 双氢睾酮体外培养人毛囊中差异表达的miR-133b对人毛乳头细胞的增殖及诱导能力的影响
- Differential Expression of miR-133b in Human Hair Follicles Treated with DHT in Vitro and Its Functional Analysis in Human Dermal Papille Cells
- 中山大学学报（医学科学版） 2021年42卷第2期页码：202-208
- 作者机构：
  
  1.中山大学附属第三医院皮肤性病科，广东广州 510630
  2.龙岗区人民医院耳鼻咽喉科，广东深圳 518172
- 作者简介：
  
  邓文佳，硕士生，研究方向：雄激素源性脱发的发病机制及治疗，E-mail:443608344@qq.com
- 基金信息：
  
  国家自然科学基金(81872542;81271769)
- DOI：
  中图分类号： R758.71
- 纸质出版日期：2021-03-20，
  
  收稿日期：2019-11-03，
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邓文佳,邓长健,韩乐等.双氢睾酮体外培养人毛囊中差异表达的miR-133b对人毛乳头细胞的增殖及诱导能力的影响[J].中山大学学报（医学科学版）,2021,42(02):202-208.

DENG Wen-jia,DENG Chang-jian,HAN Le,et al.Differential Expression of miR-133b in Human Hair Follicles Treated with DHT in Vitro and Its Functional Analysis in Human Dermal Papille Cells[J].Journal of Sun Yat-sen University(Medical Sciences),2021,42(02):202-208.
邓文佳,邓长健,韩乐等.双氢睾酮体外培养人毛囊中差异表达的miR-133b对人毛乳头细胞的增殖及诱导能力的影响[J].中山大学学报（医学科学版）,2021,42(02):202-208. DOI：

DENG Wen-jia,DENG Chang-jian,HAN Le,et al.Differential Expression of miR-133b in Human Hair Follicles Treated with DHT in Vitro and Its Functional Analysis in Human Dermal Papille Cells[J].Journal of Sun Yat-sen University(Medical Sciences),2021,42(02):202-208. DOI：

摘要

目的

探究不同浓度双氢睾酮（DHT）对毛囊生长和毛囊细胞增殖的影响及其与miR-133b表达的关系，然后进一步探究miR-133b对毛乳头细胞的增殖和诱导能力的影响。

方法

体外显微分离人毛囊，选取生长期的毛囊进行培养，显微镜下每日测量并拍摄不同浓度的DHT处理组（10

-8

、10

-7

、10

-6

和10

-5

mol/L）和对照组各组毛囊的生长长度，免疫荧光检测各组毛母质细胞Ki-67表达量评估毛囊增殖能力，qRT-PCR检测各组毛囊中miR-133b的表达量。通过Lipofectamine 2000转染miR-133b mimics及miR-133b NC至人毛乳头细胞中，CCK-8检测各组人毛乳头细胞增殖能力，qRT-PCR及Western Blot检测毛乳头细胞诱导能力指标Versican、ALP、β-catenin的mRNA及蛋白水平的相对表达量。

结果

与对照组相比，DHT 10

-5

mol/L实验组对毛囊的生长的抑制作用有统计学意义（

0.05），DHT 10

-5

mol/L实验组进入退行期的时间提前；其他组生长长度与对照组相比无统计学意义。免疫荧光及qRT-PCR显示，与对照组相比，DHT 10

-5

mol/L实验组毛母质细胞中Ki-67阳性细胞较少且毛囊中miR-133b的相对表达量明显升高，相当于对照组的（3.17±0.26）倍（

0.01）。CCK-8显示，miR-133b mimics组OD450低于miR-133b NC组（

0.05）；qRT-PCR显示，在mRNA水平，miR-133b mimics组Versican、ALP、β-catenin的相对表达量均低于miR-133b NC组（

0.001、

0.01和

0.01）。Western blot显示，在蛋白水平，miR-133b mimics组Versican、ALP、β-catenin的相对表达量同样均低于miR-133b NC组（

0.01、

0.001和

0.01）。

结论

高浓度DHT可能通过调控miR-133b的表达抑制人毛乳头细胞的增殖活性及诱导能力，最终抑制人毛囊的生长及增殖。

Abstract

Objective

To explore the effects of different concentrations of 5α-dihydrotestosterone （DHT） on growth and proliferation of human hair follicles （HFs） and their relationship with miR-133b expression， then further explore the role of miR-133b in the proliferation and inducibility of human dermal papille cells.

Methods

HFs were isolated by microdissection， then the anagen isolated HFs were cultured and divided into different concentrations of DHT treatment groups （10

-8

mol/L， 10

-7

mol/L， 10

-6

mol/L， 10

-5

mol/L） and the blank control group. The HF growth and morphology were measured and evaluated. The expression of Ki-67 in hair matrix cells and miR-133b were detected by immunofluorescence assay and qRT-PCR respectively. Lipofectamine 2000 was used to transfect miR-133b mimics and miR-133b NC into human dermal papilla cells. CCK-8 was used to assess the proliferation ability of human dermal papilla cells. qRT-PCR and Western Blot were performed to evaluate respectively the mRNA and protein levels of the markers associated with inductive ability of dermal papilla cells， such as Versican， ALP and β-catenin.

Results

Compared with the control group， no other treatment groups but the DHT 10

-5

mol/L group showed statistically significant inhibitory effect on HF growth （

0.05）. The catagen in the DHT 10

-5

mol/L group appeared earlier than that in the control group and there was no statistically significant difference in HF growth between other treatment groups and the control group. The DHT 10

-5

mol/L group showed lower percentage of Ki-67-positive cells in hair matrix cells and significantly increased relative expression of miR-133b in HFs， 3.17±0.26 times more than that in the control group （

0.01）. CCK-8 assay revealed that the OD450 value in the miR-133b mimics group was lower than that in the miR-133b NC group （

0.05）. qRT-PCR revealed that the mRNA expression levels of Versican， ALP， and β-catenin in the miR-133b mimics group were all lower than those in the miR-133b NC group （

0.001，

0.01，

0.01）. Western Blot revealed that the protein expression levels of Versican， ALP， and β-catenin in the miR-133b mimics group were also lower than those in the miR-133b NC group （

0.01，

0.001，

0.01）.

Conclusions

High concentrations of DHT may inhibit the growth and proliferation of HFs via regulating the expression of miR-133b， thus affect the proliferation and inducibility of human dermal papilla cells.

关键词

miR-133b双氢睾酮毛囊毛乳头细胞

Keywords

miR-133b5α-dihydrotestosteronehair folliclesdermal papille cells

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