自体浓缩生长因子联合毫火针激活 Wnt/β-catenin信号促进小鼠毛发生长

目的 探讨自体浓缩生长因子(concentrated growth factor, CGF)联合毫火针对雄激素性脱发(androgenetic alopecia, AGA)小鼠毛发生长的作用及机制。方法 将36只C57BL/6雄性小鼠随机分成6组(n=6),分别为CGF联合毫火针组、毫火针组、CGF组、米诺地尔阳性对照组、空白组与模型组，除空白组外，其余各组均给与睾酮涂抹建立雄激素性脱发小鼠模型，然后各组分别给予相应的治疗。肉眼评估小鼠背部皮肤毛发生长情况，HE染色后观察毛囊的形态变化及数量，Western blot及免疫组织化学检测Wnt10b、GSK-3β、β-catenin蛋白表达情况。结果 肉眼观察CGF联合毫火针可以促进小鼠毛发生长；HE染色显示毛囊数量增多，以终毛毛囊增多为著；免疫组织化学、Western blot证实Wnt10b、GSK-3β、β-catenin蛋白在上述所有组别毛囊中均有表达，且CGF联合毫火针组中Wnt10b、GSK-3β、β-catenin蛋白的表达与模型组、毫火针组、CGF组差异有统计学意义(P<0.05)。结论 CGF联合毫火针可通过激活β...     

雄激素源性脱发的治疗进展

雄激素源性脱发(AGA)是常见的毛发脱失性疾病,好发于中年人,呈慢性进行性发展.表现为头顶部毛囊变小,生长期缩短,生长期/休止期毛囊比值下降,毛发直径变小,毳毛代替终毛,出现脱发.该病发生与遗传因素、雄激素代谢失常等因素有关.在治疗上有药物、物理治疗、手术等方法.     

The progress in molecular mechanism of androgenic alopecia

雄激素性秃发(AGA)的发病涉及多数雄激素信号转导通路.AGA 患者秃发区和胡须的毛乳头细胞Ⅱ型5α-还原酶的表达高于其他部位.雄激素受体(AR) 在AGA 患者毛乳头细胞的表达高于非脱发的患者.此外,AR 共激活因子Hic-5/ARA55 在雄激素敏感部位如AGA 的秃发区和胡须的毛乳头细胞高表达.Ⅱ型5α-还原酶、AR 和Hic-5/ARA55 的表达增强可上调AGA 秃发区毛乳头细胞对雄激素的敏感性.雄激素诱导毛乳头细胞分泌转化生长因子(TGF)-β1抑制角质形成细胞生长,提示TGF-β1作为生长抑制因子是AGA 发病的决定因素之一.TGF-β2 和DKK-1 亦是雄激素诱导的毛囊上皮细胞生长抑制剂.预计更多的发病递质将被发现,进一步揭示AGA 的发病机制并寻找更好的治疗靶点.     

转化生长因子-β受体在男性雄激素性脱发患者头皮毛囊中的表达意义

目的 探讨转化生长因子-β受体(transforming growth factor-β receptor,TGF-βR)Ⅰ和Ⅱ在男性雄激素性脱发(androgenetic alopecia,AGA)患者头皮生长期毛囊中表达及其意义.方法 采用免疫组化方法对15例男性AGA患者不同头皮区域(枕部非脱发区、脱发过渡区和脱...     

Expression of BMP type ⅠA receptor in the scalp of AGA patients and its significance

目的:探讨骨形成蛋白Ⅰ型受体(BMPR-Ⅰ)A在正常人及雄激素源性脱发(androgenetic alopecia,AGA)患者头皮毛囊中的表达及其意义.方法:采用Western blot、免疫组化及RT-PCR分析8例AGA患者额顶部脱发区与枕部非脱发区毛囊及11例正常人头皮毛囊中BMPR-ⅠA蛋白及基因水平表达差异;毛囊分组培养,观察双氢睾酮(dihydrotestosterone,DHT)10-9mol/L对BMPR-ⅠA表达及毛囊生长的影响.结果:各组头皮毛囊均可见BMPR-ⅠA阳性染色.与正常毛囊相比,AGA患者脱发区毛囊阳性染色A值降低(P ＜ 0.05).Western blot结果显示,AGA患者头顶部毛囊BMPR-ⅠA含量较正常人下调,目的蛋白与内参灰度比由1.733±0.058下调为0.396±0.036(P ＜ 0.05);AGA枕部毛囊目的蛋白与内参灰度比值为1.574±0.039,与正常人组差异无统计学意义.毛囊培养DHT组与生理盐水对照组相比,目的蛋白与内参灰度比由3.441±0.172下降为0.367±0.026(P ＜ 0.05).RT-PCR结果示,AGA脱发区毛囊BMPR-ⅠA mRNA下调为正常组的53%(P ＜ 0.05),AGA枕部无明显下调;毛囊培养DHT组下调为对照组的13%(P ＜ 0.05).结论:AGA脱发区毛囊与DHT组毛囊BMPR-ⅠA表达均降低,提示BMPR-ⅠA表达下降部分是由DHT升高引起;DHT可引起的毛囊生长减慢及退行性变;BMPR-ⅠA表达下降可能是DHT升高的直接作用或者是AGA生长期与退行期毛囊比例下降的代偿机制;其参与AGA发病可能与毛干及内根鞘形成障碍引起毛发脱落有关.     

银屑病患者皮损中转化生长因子-β1及受体基因表达的研究

目的 研究银屑病患者皮损及非皮损处转化生长因子-β1(TGF-β1)、转化生长因子β受体(TGF-βRⅡ)及CD105的基因(mRNA)表达,并初步探讨其临床意义。方法 采用异硫氰酸胍法抽提皮肤组织中的RNA;采用逆转录聚合酶链反应(RT-PCR)检测皮肤组织中mRNA的表达。结果 银屑病患者皮损组织TGF-β1及TGF-βRⅡmRNA的表达均低于非皮损组织及正常人(P<0.05);非皮损组织及正常人皮肤组织中mRNA的表达量差异无显著性(P>0.05)。而银屑病皮损组织CD105的mRNA表达量高于非皮损组及正常人(P<0.05);非皮损组织和正常人皮肤组织比较P>0.05。结论 银屑病皮损中TGF-β1及TGF-βRⅡ表达降低和CD105的表达上调可能与银屑病的表皮过度增殖及真皮炎症细胞浸润有关。     

黄芪甲苷对光老化小鼠皮肤中转化生长因子-β信号通路的影响北大核心CSCD

目的探讨黄芪甲苷对皮肤光老化的保护机制。方法BALB／c小鼠分为模型组、模型＋基质组、模型＋黄芪甲苷组、正常对照组。RT—PCR测定转化生长因子β受体Ⅱ（TGF—βRⅡ）、Smad7的mRNA表达水平,免疫组化观察TGF-βRⅡ、Smad7在小鼠皮肤组织中的蛋白表达情况。结果正常对照组中,TGF-βRⅡ、Smad7的灰度值比值分别为0．5688±0．0439、0．5900±0．0585,阳性表达率分别为（53．00±4．72）％、（47．50±3．81）％;模型组中,TGF-βRⅡ、Smad7的灰度值比值分别为0．2588±0．0283、0．8637±0．0514,阳性表达率分别为（28．20±5．24）％、（82．06±2．18）％;模型＋基质组中,TGF-βRⅡ、Smad7的灰度值比值分别为0．2653±0．0456、0．8553±0．0575,阳性表达率分别为（28．74±2．28）％、（82．62±4．02）％;模型＋黄芪甲苷组中TGF-βRⅡ、Smad7的灰度值比值分别为O．3767±0．0374、0．7131±0．0410,阳性表达率分别为：（41．64±2．59）％、（64．36±2．62）％。皮肤组织中TGF-βRⅡ和Smad灰度值比值4组小鼠间比较,F值分别为80．98和736．80,TGF-βRⅡ和Smad7的阳性表达率4组小鼠间比较,F值分别为45．36和132．25,P值均〈0．01。与正常对照组相比,模型组TGF-βRⅡmRNA和蛋白表达明显降低,Smad7mRNA和蛋白表达显著升高（P值均〈0．01）。与模型组和模型＋基质组比较,模型＋黄芪甲苷组TGF-βRⅡmRNA和蛋白表达明显升高,Smad7mRNA和蛋白表达显著下调（P值均〈0．01）。模型＋基质组TGF-βRⅡ、Smad7的mRNA和蛋白表达与模型组相比差异无统计学意义（P值均〉0．01）。结论黄芪甲苷可以通过上调TGF-βRⅡ表达和下调Smad7表达而改变TGF—β通路的信号转导参与抗光老化。     

脂肪间充质干细胞对硬皮病小鼠的疗效及其对Wnt/β-catenin信号通路的影响

目的:探讨脂肪间充质干细胞(ADSCs)对硬皮病(SD)小鼠的治疗效果及其对Wnt/β-catenin信号通路的影响。方法:将40只C57BL/6J雌性小鼠随机平均分为空白组、PBS组、模型组及治疗组。模型组与治疗组通过背部皮下注射博来霉素(BLM)制备SD小鼠模型,空白组不予任何处理,PBS组注射同体积PBS溶液。造模成功后采用100μL ADSCs细胞悬液(2×1010个/L)皮下注射至治疗组小鼠背部局部皮下,每日1次,连续4周。观察各组小鼠皮肤组织病理改变,并检测其皮肤组织中羟脯氨酸含量、脾细胞中辅助性T细胞(Th)17和调节性T细胞(Treg)比例以及血清炎性因子水平。采用western blot检测皮肤组织中Wnt/β-catenin信号通路相关蛋白表达水平。结果:与空白组和PBS组相比,模型组Th17和Treg比例、血清白细胞介素(IL)-6、IL-17、Wnt2、Wnt3a及β-catenin蛋白水平均显著升高(P0.05),但IL-10水平显著降低(P0.05)。经ADSCs治疗后SD小鼠皮损组织硬化和炎症均得到明显改善,且羟脯氨酸含量显著降低(P0.05)。与模型组相比,治疗组Th17比例、血清IL-6及IL-17水平、Wnt2、Wnt3a及β-catenin蛋白表达均降低(P0.05),而Treg比例及血清IL-10水平均显著升高(P0.05)。结论:ADSCs能够通过调节T淋巴细胞和相关炎性因子分泌,改善SD小鼠机体炎症反应和皮肤组织纤维化,且该过程可能与Wnt/β-catenin信号通路有关。     

血管内皮生长因子165基因转染小鼠硬皮病皮损的实验研究

目的:探讨硬皮病基因治疗的新方法。方法:建立小鼠硬皮病动物模型后,应用电穿孔技术将血管内皮生长因子(VEGF)165真核表达质粒导入硬皮病小鼠硬化的皮下;再利用免疫组化、原位杂交法检测小鼠硬皮病皮损VEGF165的表达及观察组织病理改变。结果:①转基因后硬皮病鼠毛发生长明显增多,而未转基因硬皮病鼠毛发未见生长。②组织病理检查示转基因鼠毛囊明显增生,毛囊数每一低倍视野平均为(12.0±1.6)个,显著高于未转基因硬皮病鼠组(P<0.05)。真皮厚度增加,新生血管增加。③免疫组化、原位杂交法检测示转基因鼠表皮和真皮细胞及间质VEGF165表达明显增加。结论:电穿孔技术可将外源性基因转入小鼠硬化的皮肤,使其高表达。外源性VEGF165基因转染后,可明显促进小鼠硬化的皮肤毛发生长,已萎缩的真皮组织增生。     

丹参酮治疗雄激素性秃发大鼠模型的研究

目的：研究丹参酮对二氢睾酮（DHT）诱导的雄激素性秃发（AGA）大鼠模型的治疗作用。方法：32只Wistar雄性大鼠随机分为空白对照组、DHT造模组、丹参酮组及非那雄胺组,每组8只;空白对照组大鼠不予处理,余24只大鼠应用皮下注射5 mg/kg·d DHT的方法建立AGA大鼠模型。DHT造模组大鼠不予干预;丹参酮组大鼠予0.24 g/kg·d丹参酮进行灌胃干预,灌胃从造模的第二天开始,连续给药60 d;非那雄胺组大鼠予0.12 mg/kg·d非那雄胺进行灌胃干预,时间与丹参酮组一致。观察大鼠毛发的形态学改变及皮肤组织病理学改变,检测大鼠血清及组织的雌二醇（E2）、睾酮（T）、DHT、酸性成纤维细胞生长因子（αFGF）、胰岛素样生长因子（IGF-1）、肿瘤坏死因子（TNF-α）、转化生长因子β1（TGF-β1）水平。结果：DHT造模组大鼠背部毛发与空白对照组大鼠相比明显稀疏,部分毛发脱落,血清TNF-α、TGF-β1浓度较空白对照组明显升高（P<0.05）;丹参酮组大鼠与DHT造模组大鼠相比,背部毛发分布整齐,毛发较为浓密;丹参酮组大鼠的血清雌二醇浓度较二氢睾酮造模组显著升高（P<0.05）,血清DHT浓度较DHT造模组明显降低（P<0.05）;丹参酮组大鼠组织αFGF的浓度显著高于DHT造模组大鼠（P<0.05）。结论：丹参酮治疗AGA模型大鼠有效,其治疗AGA的机制与调节组织和循环的激素代谢水平及细胞因子作用相关。     

瘦素对雄激素性脱发小鼠模型毛囊的影响

目的：确定瘦素对雄激素性脱发小鼠毛囊的影响。方法：40只雄性C57小鼠随机分组,空白组10只不予处理,余30只背部皮下注射丙酸睾酮5 mg/kg·d进行造模。造模成功后分为3组,继续给药4周：模型组（皮下注射丙酸睾酮）、米诺地尔组（注射丙酸睾酮+外用5%米诺地尔酊）、瘦素组（注射丙酸睾酮+重组小鼠瘦素）,比较生长期与休止期毛囊数量比值（A/T）,毛囊细胞增殖与凋亡情况及雄激素受体(AR)蛋白含量。结果：(1)米诺地尔组及瘦素组较模型组脱发面积缩小;(2)米诺地尔组与瘦素组较模型组的生长期/休止期毛囊数比值、Ki67表达均明显增加（均P＜0.05）,两者在米诺地尔组与瘦素组之间比较均无明显统计学差异（均P＞0.05）;(3)毛囊凋亡数量及AR水平在米诺地尔组与瘦素组间比较均无明显统计学差异（均P＞0.05）。结论：皮下注射瘦素对雄激素性脱发模型小鼠有明确治疗作用,且效果与外用5%米诺地尔溶液作用相当。     

Occipital involvement in female pattern hair loss: histopathological evidences

Background Female pattern hair loss (FPHL) is characterized by diffuse thinning of hair in the frontal and parietal areas of the scalp, and preservation of the frontal hairline is the norm. Hair on the occipital scalp is thought to be preserved. Objective To investigate whether the occipital area is involved in FPHL or whether there is a diffuse type of FPHL. Methods Forty female patients who had complained about hair loss for more than a year and were diagnosed with FPHL according to the Ludwig classification were included. Two punch biopsies from both the midscalp and the occiput were taken. Histological sections were prepared horizontally and stained with haematoxylin and eosin. Terminal follicles, vellus like follicles, anagen, telogen, catagen follicles, hair bulbs and telogen germinal units were counted in two sections of the upper dermis and the dermal‐subcutaneous junction. If the terminal/vellus ratio was lower than 4:1, the diagnosis of androgenetic alopecia (AGA) was made. When the ratio was between four and seven to one, AGA was suspected. Results While 29 of 40 patients (72.5%) had findings consistent with AGA on the midscalp, 11 of 40 (27.5%) displayed signs of suspected AGA. Ten of 40 patients (25%) had AGA involving the occiput. Conclusion The involvement of the occipital scalp is significant in FPHL. In some patients, this situation may be so apparent that clinically visible alopecia is seen. However, in other patients, it may also present only as thinning.     

Effect of caffeine and testosterone on the proliferation of human hair follicles in vitro

BACKGROUND: Androgenetic alopecia (AGA) is a common problem in men of all ages, affecting approximately 50% at 50 years of age. The underlying cause is an androgen-dependent miniaturization of genetically predetermined hair follicles. Here, the hair organ culture model was used to investigate the effects of testosterone and caffeine; the latter being a promising candidate for hair growth stimulation. METHODS: Hair follicles from 14 biopsies, taken from the vertex areas from male AGA patients, were cultivated for 120-192 h in vitro with normal William's E medium (control) or William's E medium containing different concentrations of testosterone and/or caffeine. Hair shaft elongation was measured daily and at the end of cultivation, cryosections of follicles were stained with Ki-67 to evaluate the degree and localization of keratinocyte proliferation. RESULTS: Significant growth suppression was found in hair follicles treated with 5 microg/ml testosterone. This was counteracted by caffeine in concentrations of 0.001% and 0.005%. Moreover, caffeine alone led to a significant stimulation of hair follicle growth. These results were confirmed immunohistochemically by Ki-67 staining. CONCLUSIONS: Androgen-dependent growth inhibition of ex vivo hair follicles from patients suffering from AGA was present in the human hair organ culture model, a constellation which may serve for future studies to screen new substances against androgen-dependent hair loss. Caffeine was identified as a stimulator of human hair growth in vitro; a fact which may have important clinical impact in the management of AGA.     

Treatment of androgenetic alopecia by exosomes secreted from hair papilla cells and the intervention effect of LTF

Background

Androgenetic alopecia (AGA) is the most common cause of chronic progressive hair loss in men, and AGA has a severe negative impact on the quality of life and physical and mental health of patients.

Methods

Four female C57BL/6 mice were isolated from DP cells in culture (≤4 generations) after stimulation of DPC proliferation by herbal concentrations obtained by the CCK-8 method, and exosomes were isolated by differential centrifugation at low temperature. Testosterone propionate and topical hair removal treatments were used together to establish the C57BL/6 mouse AGA model, which was treated with LTF, 5% minoxidil, and LTF-DPC-EXO, respectively. ELISA was used to detect serum hormone levels, in vivo tracing was used to observe dynamic changes in exosomes, H&E staining showed changes in mouse hair follicle tissue, and (q) RT-PCR and WB were used to detect dorsal skin VEGF, AKT1, and CASP3 expression in dorsal skin tissues.

Results

Hair regeneration was significant in the LTF group, minoxidil group, and LTF-DPC-EXO group mice, and the hair growth was only seen in the local skin in the model group. The hormone T in all treatment groups was lower than that in the model group, and e2 was higher than that in the model group. (q) RT-PCR and western blot showed that VEGF and AKT1 expressions were upregulated and Caspase3 expression was downregulated in the skin sections of mice in the treatment groups.

Conclusion

DPC-EXO obtained through LTF may activate AKT1 and VEGF in the PI3K/AKT signaling pathway to inhibit CASP3, thereby protecting DPC to restore the hair growth.     

Kenogen in female androgenetic alopecia. A longitudinal study

BACKGROUND: Kenogen indicates the physiological interval of the hair cycle in which the hair follicle remains empty after the telogen hair has been extruded and before a new anagen hair emerges. Kenogen frequency and duration are greater in men and women with androgenetic alopecia (AGA). OBJECTIVE: To study the relationship of kenogen with female AGA. METHODS: A woman with AGA, studied 14 years before, was re-examined for 2 years by the phototrichogram technique. Kenogen was identified when telogen lasted 3 months and shedding left the follicle empty. RESULTS: Kenogen lasted 1-5 months, the longest duration affecting cycles of vellus hairs, and involved more hair follicles (16 vs. 8%) than in the first observation. Cycles of vellus hairs increased in number from 4 to 13%, and cycles with a normal succession of phases decreased from 60 to 32%. CONCLUSIONS: The number of kenogen phases increased in parallel with vellus hairs and the diminished number of normal hair cycles, features that mark AGA aggravation. Kenogen seems to be related also to the progression of female AGA.     

Transplants from balding and hairy androgenetic alopecia scalp regrow hair comparably well on immunodeficient mice

Human hair follicles were grafted onto 2 strains of immunodeficient mice to compare the regeneration potential of vellus (miniaturized, balding) and terminal (hairy, nonbalding) follicles from males and a female exhibiting pattern baldness. Each mouse had transplants of both types of follicles from a single donor for direct comparison. Grafted follicles from 2 male donors resulted in nonsignificant differences in mean length (52 mm vs 54 mm) and mean diameter (99 microm vs 93 microm) at 22 weeks for hairs originating from balding and hairy scalp, respectively, corresponding to 400% versus 62% of the mean pretransplantation diameters. Follicles from the female donor transplanted to several mice also resulted in nonsignificant differences in length (43 mm vs 37 mm) for hairs from balding and hairy scalp, respectively, during a period of 22 weeks. The mean diameter of the originally vellus hairs increased 3-fold, whereas the terminal hairs plateaued at approximately 50% of pretransplantation diameter, resulting in a final balding hair volume double that of the nonbalding hairs. This report shows that miniaturized hair follicles of pattern alopecia can quickly regenerate once removed from the human scalp and can grow as well as or better than terminal follicles from the same individual.     

A bald statement — Current approaches to manipulate miniaturisation focus only on promoting hair growth

Hair plays a large part in communication and society with its role changing through time and across cultures. Most people do not leave the house before combing their hair or shaving their beard and for many hair loss or irregular hair growth can have a significant impact on their psychological health. Somewhat unsurprisingly, according to GMR Data, today’s global hair care industry is worth an estimated $87 Billion, with hair loss estimated at $2.8 Billion. Considering that no current hair loss‐related products can completely reverse hair loss, it is reasonable to believe this market could expand significantly with the discovery of a comprehensive therapy. As such, a great deal of research focuses on overcoming hair loss, and in particular, a common form of hair loss known as androgenetic alopecia (AGA) or male pattern baldness. In AGA, hair follicles miniaturise in a large step change from a terminal to a vellus state. Within this viewpoint article, we discuss how influx and efflux of cells into and out from the dermal papilla (DP) can modulate DP size during the hair cycle. As DP size is positively correlated with the size of the hair fibre produced by a follicle, we argue here that therapies for treating AGA should be developed which can alter DP size, rather than just promote hair growth. We also discuss current therapeutics for AGA and emphasise the importance of using the right model systems to analyse miniaturisation.     

Quantification of hair follicle parameters using computer image analysis: A comparison of androgenetic alopecia with normal scalp biopsies

Computer image analysis enables large numbers of hairs to be measured in an automated fashion. In this study, we examined horizontal scalp biopsies from 10 patients with a histological diagnosis of androgenetic alopecia and 10 normal control subjects. The density of hair follicles and the ratio of terminal to vellus hairs were determined. Hair shaft, hair canal and hair follicle diameter, inner root sheath width and outer root sheath area were measured using the Chromatic Colour Image Analysis program. This study showed a statistically significant progressive decrease in size of hair canal diameters from normal terminal hairs (85.93 ± 10.07 μm) through to androgenetic alopecia terminal (68.83 ± 13.60 μm) and vellus hairs (28.67 ± 5.60 μm). This pattern is also seen with hair follicle diameters; normal terminal (268.41 ± 24.88 μm), androgenetic alopecia terminal (236.34 ± 17.23 μm), and vellus hairs (130.88 ± 19.96 μm). Outer root sheath areas, hair shaft diameters and ratio of terminal to vellus hairs were significantly larger in normal (18500 ± 4222 μm²; 82.71 ± 13.79 μm; 36:1; respectively) compared with androgenetic alopecia scalp biopsies (8403 ± 3322 μm²; 61.11 ± 14.42 μm; 3:1; respectively), whereas inner root sheath width and density did not vary significantly. Computer image analysis can be adapted for use in clinical trials where large numbers and objectivity are critical in determining the efficacy of hair growth promoters.     

The diagnosis of androgenetic alopecia in children: Considerations of pathophysiological plausibility

Androgenetic alopecia (AGA), one of the most common causes of hair loss in men and women, is an infrequent cause of alopecia in children. In AGA, patients generally start noticing hair thinning after the onset of puberty due to progressive miniaturisation of the hair follicle which leads to vellus transformation of terminal hair. However, the occurrence of prepubertal AGA has rarely been reported in the literature. The pathophysiology of AGA is tightly linked to androgen hormones; prepubertal children do not usually produce significant amounts of adrenal or gonadal androgens. When it does occur, an underlying abnormality should be suspected. Secondary causes of AGA must be excluded when evaluating a patient before the appearance of puberty. Premature puberty, polycystic ovarian syndrome and other causes of hyperandrogenism can present with hair loss in an androgenetic pattern. This article reviews the normal physiology of androgen hormones and their role in the pathophysiology of childhood AGA.