Hydroxysafflor yellow A (HSYA) is an effective chemical component isolated from Chinese herb Carthamus tinctorius L. In present study, we aimed to evaluate the effects of HSYA on D-galactose- (D-gal-) induced aging in mice, and to elucidate the underlying mechanism. Male C57BL/6 mice were intraperitoneal injection of D-gal and HSYA for 8 weeks. The body weight gain, spleen and thymus coefficients were determined. Levels of super dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) in serum and liver were measured using commercial kits. Pathological changes and the SA-β-Gal activity in liver tissues were detected by hematoxylin and eosin and SA-β-Gal staining. The expression levels of p16, CDK4, CDK6 and phosphorylation levels of Retinoblastoma (Rb) were detected by immunohistochemistry and western blot analysis. mRNA levels of genes regulated by p16-Rb pathway were determined by quantitative real-time PCR. In vivo, HSYA improved the aging changes including body weight, organ index and antioxidant status such as activities of SOD, CAT, GSH-Px and MDA in D-gal treated aging mice. HSYA also dramatically attenuated pathologic changes of aging liver tissues induced by D-gal. Furthermore, HSYA significantly decreased the mRNA and protein level of cyclin-dependent kinase inhibitor p16, followed by increasing CDK4/6 protein expression and decreasing the phosphorylation of Retinoblastoma (pRb) which up-regulated the expression of downstream genes CCNE1, CCNA2, P107 and MCM4. Collectively, these data indicated that HSYA could ameliorate aging, especially hepatic replicative senescence resulting from D-gal, the mechanism could be associated with the suppression of p16-Rb pathway.

羟基红花黄A（HSYA）是从中草药红花（Carthamus tinctorius）中分离出来的有效化学成分。L.在本研究中，我们旨在评估HSYA对D-半乳糖（D-gal-）诱导的小鼠衰老的影响，并阐明其潜在机制。雄性C57BL / 6小鼠腹膜内注射D-gal和HSYA达8周。确定体重增加，脾脏和胸腺系数。使用商业试剂盒测量了血清和肝脏中超歧化酶（SOD），过氧化氢酶（CAT），谷胱甘肽过氧化物酶（GSH-Px）和丙二醛（MDA）的水平。用苏木精和曙红和SA-β-Gal染色检测肝组织的病理变化和SA-β-Gal活性。通过免疫组织化学和蛋白质印迹分析检测p16，CDK4，CDK6的表达水平和视网膜母细胞瘤（Rb）的磷酸化水平。通过定量实时PCR确定由p16-Rb途径调控的基因的mRNA水平。在在体内，HSYA改善了D-gal治疗的衰老小鼠的衰老变化，包括体重，器官指数和抗氧化剂状态，例如SOD，CAT，GSH-Px和MDA的活性。HSYA还显着减轻了D-gal引起的衰老肝组织的病理变化。此外，HSYA显着降低了细胞周期蛋白依赖性激酶抑制剂p16的mRNA和蛋白水平，随后增加了CDK4 / 6蛋白的表达并降低了视网膜母细胞瘤（pRb）的磷酸化，从而上调了下游基因CCNE1，CCNA2，P107和MCM4。总的来说，这些数据表明HSYA可以改善衰老，尤其是D-gal引起的肝复制性衰老，其机制可能与抑制p16-Rb途径有关。