Method and composition for treatment of hair loss
A composition and application technology, applied in chemical instruments and methods, biochemical equipment and methods, drug combinations, etc., can solve problems such as little understanding of effects
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Embodiment 1
[0042] Example 1: Blockade of Notch signaling in hair follicle progeny leads to delayed return to anagenre-entry
[0043] Previous studies on the role of Notch signaling in the hair follicle cycle using knockout of the Rbpj gene via Krt14-Cre resulted in death of neonatal mice due to severe defects in epidermal barrier function. In order to avoid the lethality of neonatal mice caused by the removal of Notch signaling in the basal epidermis, this example uses Tgfb3-Cre to knock out Pofut1 in epidermal epithelial and hair follicle cell progeny (Lin HY et al., PLoS One 6:e15842, 2011 ).
[0044]Generation of Tgfb3-Cre (Tgfb3-Cre+ / wt), floxed Pofut1 (Pofut1fx / fx) and floxed Hes1 (Hes1fx / fx) mice has been previously described (Shi S, Stanley P., Proc Natl Acad Sci US A100:5234 -5239, 2003; Yang LT, Li WY, Kaartinen V. 2008. Genesis 46:112-118. ImayoshiI et al, Development 135:2531-2541, 2008). Crossing Pofut1fx / fx mice with Tgfb3-Cre mice produced heterozygous Tgfb3-Cre+ / wt;Pofut...
Embodiment 2
[0050] Example 2: Notch signaling regulates proper proliferation and cell death of hair follicle stem / progenitor cells during the hair follicle cycle
[0051] Next, this example investigates the underlying mechanisms responsible for the smaller bulges / hair embryos observed in Pofut1 / Tgfb3-Cre mice by analyzing the state of cell proliferation and apoptosis in the hair cycle.
[0052] The control group and Pofut1 / Tgfb3-Cre mice were marked with BrdU-marked at different hair cycle stages (from senescence to late anagen), and BrdU immunostaining was performed. The results of BrdU staining and quantitative analysis showed that in the hair follicles of mutant mice, the cell proliferation in the hair germ at P24 (early anagen) and in the hair follicle bulb at P30 (late anagen) was significantly higher than that in the control group The sample is greatly reduced ( figure 2 A). Notably, BrdU staining was observed in ORS of mutant follicles regressing (at P20 extinction), and in the ...
Embodiment 3
[0055] Example 3: Blockade of Notch signaling in hair follicle progeny leads to DNA damage response and lack of DNA repair mechanisms
[0056] Delayed reentry into anagen during hair cycle transition and dysregulation of cell proliferation / apoptosis observed in Pofut1 / Tgfb3-Cre mice prompted in this example to examine whether loss of Notch signaling affects mutant hair follicles Some stress responses, such as the DNA damage response or cellular senescence, are caused in
[0057] For senescence-associated β-galactosidase assays, cryosections were fixed in 0.2% paraformaldehyde for 20 minutes on ice and β-galactosidase activity was visualized using X-gal staining following standard procedures. After X-gal staining, sections were counterstained with eosin. Aging-associated β-galactosidase assays did not reveal senescence in hair follicle cell progeny. However, more γ-H2AX staining signal was observed in Pofut1 / Tgfb3-Cre hair follicle cells (compared to control hair follicle cel...
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