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目的:探讨哈蟆油(OR)对D-半乳糖所致雌性衰老大鼠子宫组织细胞增殖调控因子p21和cyclin D1基因表达的影响,进一步探讨OR延缓雌性大鼠生殖器官衰老机制。方法:SPF级SD雌性青年大鼠40只随机分为模型组(D-gal组)、维生素E(VE组)、哈蟆油高剂量组(OR-H组)、中剂量组(OR-M组)、低剂量组(OR-L组),每组8只,D-半乳糖颈背部皮下注射42 d,建立亚急性衰老模型。另取雌性青年大鼠8只,同样部位每日注射生理盐水,作为空白组。第15天开始灌胃给药,给药时间28 d。给药结束后,实时荧光定量PCR(q RT-PCR)法检测衰老大鼠子宫组织p21和cyclin D1基因的表达情况。结果:雌性衰老大鼠各组子宫组织q RT-PCR结果显示p21和cyclin D1基因表达相比均有显著的统计学意义(P<0.01)。D-gal组p21基因表达与Normal组比较升高,差异有显著性(P<0.01)。OR各剂量组p21基因表达与D-gal组比较下降,差异有显著性(均P<0.01),OR-L组作用较为明显。D-gal组cyclin D1基因表达与Normal组比较降低,差异有显著性(P<0.01);OR各剂量组cyclin D1基因表达与D-gal组比较均升高,差异有显著性(均P<0.01),表达随着OR剂量增加而升高。结论:OR可降低雌性衰老大鼠子宫组织细胞增殖负性调控因子p21基因表达,提高正性调控因子cyclin D1基因表达,促进衰老雌性大鼠子宫细胞增殖。OR延缓雌性生殖器官衰老作用可能通过调控子宫组织p21-cyclin D1信号通路发挥作用。
Objective: To investigate the effect of Rana oil on the gene expression of p21 and cyclin D1 in the uterus of aging female rats induced by D-galactose, and to further explore the mechanism of OR delaying the aging of reproductive organs in female rats. Methods: Forty SPF female SD rats were randomly divided into three groups: model group (D-gal group), vitamin E group (VE group), high dose group of Toad oil (OR-H group) Group), low dose group (OR-L group), 8 rats in each group. D-galactose was subcutaneously injected into the back of the neck for 42 days to establish a sub-acute aging model. Another 8 young female rats, the same site daily injection of saline, as a blank group. The first 15 days began gavage, dosing time 28 d. After the administration, real-time quantitative PCR (q RT-PCR) was used to detect the gene expression of p21 and cyclin D1 in the uterus of aging rats. Results: q RT-PCR results showed that the expression of p21 and cyclin D1 were significantly different (P <0.01). The expression of p21 gene in D-gal group was higher than that in Normal group, the difference was significant (P <0.01). Compared with D-gal group, the expression of p21 gene in each dose group decreased significantly (all P <0.01), and the effect of OR-L group was more obvious. The cyclin D1 gene expression in D-gal group was significantly lower than that in Normal group (P <0.01), and the cyclin D1 gene expression in D-gal group was significantly higher than that in D-gal group (all P < 0.01). The expression increased with the increase of OR dose. Conclusion: OR can reduce the expression of p21 gene in the uterus of uterus in aging rats, increase the expression of cyclin D1, and promote the proliferation of uterine cells in aging female rats. The role of OR in delaying female reproductive organ aging may play a role in the regulation of p21-cyclin D1 signaling in uterine tissue.