肝细胞生长因子对小鼠触须毛囊体外培养的研究

近年研究已经发现许多细胞因子和生长因子对毛发生长有调控作用^[1,2].尤其是肝细胞生长因子(HGF)对毛囊体外培养有明显的促进作用.我们应用C₅₇BL/6J小鼠触须毛囊体外培养模型,对HGF促毛发生长作用进行研究,现将研究结果报道如下.     

小鼠触须毛囊体外培养的研究

目的 为了建立较理想的小鼠触须毛囊体外培养模型,对不鼠触须毛囊体外培养方法进行了研究。方法 通过小鼠触须毛囊体外无血清培养基的方法,对不同鼠龄及不同生长周期的毛囊进行了研究。结果 ①毛囊在Ｗｉｌｌｉａｍｓ－Ｅ加Ｌ－谷酰胺培养基上生长平均达８天;②通过对比不同阶段的生长期毛囊发现生长期－Ⅱ毛囊生长力明显优于其它各期毛囊;③对不同鼠龄毛囊体外增培养生长状况的研究发现出生３５天和６５天鼠优于出生７天乳鼠     

甲状旁腺激素对小鼠毛发周期和毛发生长的影响

以C57BL6小鼠作为动物模型,观察甲状旁腺激素(PTH)的多肽片段PTH(7～34)和PTH(1～34)对毛发周期和毛发生长的影响。用松香/蜡混合物拔毛法诱导C57BL6小鼠毛发由休止期进入生长期,拔毛同时腹腔注射PTH(7～34),持续19天,观察毛发周期的变化。结果,PTH(7～34)组小鼠在拔毛后第5天毛发进入生长期,比对照组提前24h左右;至第19天,对照组小鼠多数毛发进入退行期或休止期,而PTH(7～34)组仍有大部分毛发维持在生长期。C57BL6乳鼠触须毛囊体外培养结果,加PTH(7～34)10-6mol/L浓度组和10-7mol/L浓度组毛干生长速度显著比对照组加快(P<0.01);而PTH(1～34)10-9mol/L浓度组毛干生长明显慢于对照组(P<0.01)。提示PTH的不同片段对毛发生长有不同影响,PTH(7～34)可延长毛发生长期,促进小鼠毛发生长,而PTH(1～34)则对毛发生长有抑制作用。     

甲状腺素对小鼠毛发生长周期的影响

利用Ｃ５７ＢＬ６小鼠作动物模型,观察甲状腺素对毛发生长周期的影响。选择毛发处于休止期的Ｃ５７８ＢＬ６小鼠,用不同剂量的左旋 腺素钠连续灌胃２８ｄ,并以松香／蜡混合物对小鼠背部皮肤进行拔毛诱导生长期作对照。     

55种中药对小鼠触须毛囊体外培养生物学特性的研究

应用小鼠触须毛囊体外培养模型,通过培养毛囊的形态学和生长速度以及毛囊对3H-TdR掺入率的观察,对55种单味中药的水煎剂或其单体进行了实验研究。结果发现女贞子、白芷、白芨、荆芥、黄芪、潼蒺藜、甘草酸对体外培养的小鼠触须毛囊有明显的促生长作用,而补骨脂、白蔹、防风、大黄、丹参、白芍、槟榔、茜草等对体毛囊生长有明显抑制作用。对女贞子的主要成分齐敦果酸进行研究发现,其促毛囊生长作用呈浓度依赖性。该研究排除了体内实验中诸多影响因素,具有实验周期短,数据稳定可靠,为从中药中研制有效的促进毛发生长或脱毛制剂提供了重要依据。     

环孢素A对小鼠触须毛囊体外培养的研究

目的：临床发现使用环孢素Ａ的患者毛发多浓密或出现多毛,但其局部外用治疗脱发的疗效报告不一。本研究就环孢素Ａ对毛囊生物学特性的影响进行研究。方法：我们用小鼠触须毛囊体外培养的模型,通过对体外培养毛囊的生长速度、毛球部形态学改变及毛囊＾３Ｈ－ＴｄＲ掺入试验,对环孢素Ａ促毛发生长的作用进行研究。结果：本研究发现,与阴性对照组相比,０．０１μＭ环孢素Ａ使体外培养的毛囊生长速度加快（Ｐ〈０．０５）,并持续生长达１０天以上。行囊对＾３Ｈ－ＴｄＲ掺入率的试验与上述结果一致。毛球部形态学研究发现,培养第１０天,环孢素Ａ组毛球部仍然呈现长期样毛球部的形态学表现,而长压定组和阴性对照组则已表现为退行／休止期外观。结论：本研究证实了环孢素Ａ对毛发生长有明显的促进作用。该作用主要表现在两个方面：①使毛囊生长期延长;②同时对毛囊细胞的增     

血管内皮细胞生长因子对小鼠触须毛囊体外培养的研究

ＶＥＧＦ（血管内皮细胞生长因子）是新近发现于毛囊的生长因子。我们应用新近建立的Ｃ５７Ｂ－Ｌ／６Ｊ小鼠触须嘴毛囊体外培养模型。对ＶＥＧＦ促毛囊生长的生物学特性进行研究。结果发现,ＶＥＧＦ有明显促毛发生长作用,在培养的第２天,ＶＥＧＦ组毛发生长长度就明显优于阴性对照组。ＶＥＧＦ组毛发生生长度就明显优于阴性对照组。ＶＥＧＦ毛囊生长作用随其浓度增高而增强,当浓度为１０ｎｇ／ｍｌ时促生长作用达高峰。ＶＥＧＦ     

不同中药色素对C57BL/6小鼠触须毛囊体外培养的研究

目的 研究不同中药色素对小鼠体外毛囊生长的影响和作用机制.方法 分离小鼠触须毛囊,采用不同中药色素分多个浓度分别培养毛囊.在倒置显微镜下观察毛囊组织的形态学变化并测定毛干的长度,2次/d拍照记录;噻唑蓝(MTT)比色法检查毛囊细胞活力.结果 ①与阴性对照组比较,3μg/mL及10μg/mL的姜黄素、30μg/mL的紫草...     

Clues that mitochondria are involved in the hair cycle clock: MPZL3 regulates entry into and progression of murine hair follicle cycling

The molecular nature of the hair cycle clock (HCC), the intrinsic oscillator system that drives hair follicle (HF) cycling, remains incompletely understood; therefore, all relevant key players need to be identified. Here, we present evidence that implicates myelin protein zero-like 3 (MPZL3), a multifunctional nuclear-encoded mitochondrial protein known to be involved in epidermal differentiation, in HCC regulation. By analysing global Mpzl3 knockout (−/−) mice, we show that in the absence of functional MPZL3, mice commence HF cycling with retarded first catagen-telogen transition after normal postnatal HF morphogenesis. However, Mpzl3 −/− mice subsequently display strikingly accelerated HF cycling, i.e. a precocious telogen-to-anagen transition during the second hair cycle, compared to controls, suggesting that MPZL3 inhibits anagen entry. We also show that intrafollicular MPZL3 protein expression fluctuates in a hair cycle-dependent manner. In telogen HFs, MPZL3 is localized to the secondary hair germ, an epicentre of hair cycle regulation, where it partially co-localizes with P-cadherin. In early anagen HF, MPZL3 is localized immediately distal to the proximal hair matrix. These findings introduce the novel concept that mitochondria are more actively involved in hair cycle control than previously recognized and that MPZL3 plays a central role in the HCC.     

Characterization of dermal type I collagen of C3H mouse at different stages of the hair cycle

Hair follicles develop or regress in accordance with the hair cycle. In this study, we partially characterized fibrillar type I collagen, the predominant component in the dermis, at two stages of the hair cycle: anagen and telogen. Skin samples were obtained from the backs of two groups of 11-week-old C3H mice: one at anagen stage induced by shaving and the other at telogen stage. The amount of neutral salt-soluble (newly synthesized) collagen obtained from anagen skin was about twofold that from telogen skin, while the level of acid-soluble collagen was not significantly different between the two groups. The degree of lysine hydroxylation of pepsinized type I collagen obtained from anagen skin was significantly higher than that in telogen (5.0% higher in alpha1 chain, and 15.6% higher in alpha2 chain). Proline hydroxylation at the anagen stage was also slightly higher than in the telogen stage. Two major collagen cross-links were found in both groups of skin; dehydro-hydroxylysinonorleucine and dehydro-histidinohydroxymerodesmosine. The concentration of the latter, a complex tetravalent cross-link, was significantly lower in anagen skin when compared with telogen skin (mean +/- SD 0.64 +/- 0. 07 vs. 0.78 +/- 0.06 mol/mol collagen). The former showed no significant difference between the two groups. In addition, a significant amount of lysyl-aldehyde (a cross-link precursor) was found in anagen (0.16 +/- 0.02 mol/mol collagen), while it was 0.12 mol/mol collagen in telogen. These results indicate that the remodelling of collagen is more active in anagen skin than in telogen, and that characteristic post-translational modifications of dermal collagen seen in anagen may play a part in facilitating an environment around hair follicles for their migration and growth.     

Effect of Plantago asiatica L. extract on the anagen phase in human hair follicle dermal papilla cells

Background

The hair growth cycle consists of the anagen, catagen, and telogen phases, and hair follicle dermal papilla (HDP) cells of human hair play a role in the initiation and maintenance of the anagen phase. Reduction in HDP cells contributes to hair loss; however, the limited treatment options are associated with negative side effects. Therefore, a naturally derived substance with hair loss-preventing properties is needed.

Aim

We investigated the hair growth-stimulating activities of Plantago asiatica L. extract (PAE) and its molecular mechanism in HDP cells.

Methods

Cell proliferation was determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide solution. Relative mRNA and protein expression levels of hair growth factors were determined using quantitative real-time polymerase chain reaction and western blotting, respectively. Additionally, a tube formation assay was performed in human umbilical vein endothelial cells (HUVEC).

Results

Plantago asiatica L. extract significantly increased the cell proliferation and expression of hair growth factors, including keratinocyte growth factor (KGF), vascular endothelial growth factor (VEGF), fibroblast growth factor 2 (FGF2) and MYC, in HDP cells. Moreover, PAE led to the accumulation of β-catenin by promoting the phosphorylation of glycogen synthase kinase-3 beta (GSK-3β) at Ser9 and cAMP response element-binding protein (CREB) at Ser133 via phosphorylation of extracellular signal-regulated kinase (ERK) (Thr202/Tyr204). PAE also increased tube formation in HUVECs, which promoted angiogenesis for the anagen phase.

Conclusions

Plantago asiatica L. extract amplified tube formation and production of growth factors (KGF, VEGF) via the activation of GSK-3β/β-catenin and mitogen-activated protein kinase (MAPK)/CREB signaling pathways, demonstrating its potential to safely promote hair growth by inducing the anagen phase.     

活血补肾合剂对C57BL/6小鼠毛发生长的影响

目的从微观结构方面探讨活血补肾合剂促进小鼠毛发生长的机理.方法用C57BL/6小鼠建立动物模型,对120只小鼠以松香/蜡混合物(1:1)拔毛,诱导其毛发由休止期进入生长期,并随机分为4个组,即实验组、对照组、模型组和空白组.从造模后第一天开始灌药,每天观察各组皮肤色泽变化,并分别于造模后第4、11、17天每组处死10只小鼠,观察拔毛部位的毛囊和血管情况.结果活血补肾合剂第4天时显示出促进血管新生(P＜0.05),在第11天时表现出促进毛囊新生(P＜0.05),第17天表现出促进毛囊成熟的作用(P＜0.05).结论活血补肾合剂可能是通过促进血管新生,改善局部血液循环达到促进毛发生长的目的.     

雷帕霉素对HaCaT细胞IL-6、CXCL8及VEGF表达的影响及其与PI3K/Akt/mTOR信号通路的关联

目的:通过研究Rapmycin与LY294002对人永生化角质形成细胞(HaCaT细胞)白细胞介素(IL)-6、趋化因子蛋白(CXCL)8及血管内皮生长因子(VEGF)表达及其PI3K/Akt/mTOR信号通路的影响,探寻银屑病潜在治疗药物。方法:收集我院皮肤科就诊并确诊为银屑病的患者62例;并招募健康志愿者34人,分别取血检测血清中IL-6、CXCL8及VEGF水平。分别对人表皮永生化角质细胞(HaCaT)给与Rapmycin、LY294002及肿瘤坏死因子(TNF)-α培养12 h,后用试剂盒检测细胞上清中的IL-6、CXCL8及VEGF水平,用Q-PCR检测细胞中IL-6、CXCL8及VEGF的基因表达水平,用western检测Akt、S6K及4EBP1总蛋白和磷酸化蛋白的表达水平。结果:与健康志愿者相比,银屑病患者体内的IL-6、CXCL8及VEGF水平均显著上升(P<0.05);肿瘤坏死因子(TNF)-α显著提高了细胞上清中IL-6、CXCL8及VEGF水平(P<0.05)、细胞中IL-6、CXCL8及VEGF基因水平(P<0.05),并促进了细胞中Akt、...