This study aimed to explore the effect of glucose on the proliferation, migratory capacity, and programmed cell apoptosis of in vitro-cultured secondary dermal papilla cells (DPCs) derived from Yanshan cashmere goats. Skin samples were collected from healthy male Yanshan cashmere goats aged about 6 months old. Secondary hair follicles were then harvested from this tissue, and the derived DPCs were subjected to a process of extraction and purification. A growth curve was established after cell counting. The purified DPCs population was then subjected to a random distribution across 5 groups, each containing 6 replicate cultures. Glucose solutions of 0 (control group), 25, 50, 100 and 200 μg·mL-1 were added to the culture medium, respectively. The proliferation ability of DPCs was evaluated by the CCK-8 method at 2, 12, 24 and 48 h of culture. A cell scratch assay was conducted to evaluate the migratory capacity of DPCs, while the extent of apoptosis was quantified via flow cytometry following Annexin V-FITC/PI dual staining. The transcriptional responses to glucose exposure were assessed by real-time qPCR, measuring the mRNA abundance of key proliferation markers (PCNA, CCND1) and apoptosis regulators (Bcl-2, Bax, Caspase-3, FGF21, FGF5, P53). Furthermore, Western blot analysis was performed to determine the effect of glucose on the expression of corresponding functional proteins, including the proliferation- and anti-apoptosis-related factors (PCNA, CCND1, Bcl-2), as well as the pro-apoptotic executors (Bax, Caspase-3). The results demonstrated that at the 12-hour time point, both 100 and 200 μg·mL-1 glucose treatments significantly enhanced the proliferation and migration rates of DPCs (P<0.05), while concurrently suppressing apoptosis (P<0.05). Further investigation revealed these concentrations markedly upregulated the mRNA expression of proliferation- and anti-apoptosis-related genes (PCNA, CCND1, Bcl-2) (P<0.05), and downregulated transcripts of key apoptosis-associated genes (Bax, Caspase-3, FGF21, FGF5, P53) (P<0.05). At the protein level, the 100 μg·mL-1 treatment significantly elevated the expression of proliferation and anti-apoptotic markers (PCNA, CCND1, Bcl-2) and reduced pro-apoptotic Bax protein (P<0.05). Conversely, the 200 μg·mL-1 glucose group specifically induced a pronounced downregulation of the critical apoptosis executor Caspase-3 (P<0.05). In conclusion, the pro-proliferative and anti-apoptotic effects observed in cashmere goat DPCs following treatment with 100 and 200 μg·mL-1 glucose are mediated through a dual regulatory mechanism. This involves the coordinated transcriptional and translational upregulation of key proliferative markers (PCNA, CCND1) and the anti-apoptotic protein Bcl-2, concurrent with the suppression of specific apoptosis-related transcripts (FGF21, FGF5, P53) and both the gene and protein expression of pro-apoptotic mediators Bax and Caspase-3.
| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |