目的探讨人参皂苷Rg1对D.半乳糖致衰老大鼠脾组织结构与功能的影响及其机制。方法sD大鼠随机分为正常对照组、衰老模型组(D-半乳糖120mg/kg,qd×42d)、Rgl干预组(建模,第15天起Rg120mg/kg,qd×28d)、Rgl对照组(0.9%氯化钠注射液+Rgl)。取脾脏测定脾指数,石蜡切片观察脾脏显微形态,B-半乳糖苷酶(SA—β—Gal)染色检测脾细胞衰老,CCK-8检测脾细胞对刀豆蛋白A刺激的增殖能力,ELISA检测IL-2、IL-6和晚期糖基化终产物(AGEs)含量,流式细胞术检测细胞活性氧(ROS),酶法检测丙二醛(MDA)及超氧化物歧化酶(SOD)含量,Western blot检测细胞衰老相关蛋白P53、P21及Rb的表达。结果与衰老模型组比较,Rgl干预组大鼠脾指数、脾脏白髓面积比及脾细胞增殖能力提高(P〈0.05);脾细胞分泌IL-2、IL-6水平和SOD活性明显增强(P〈0.01);脾细胞的SA-B-Gal阳性率、ROS和MDA含量下降(P〈0.01);AGEs下降(P〈0.05);P53、P21及RB蛋白表达显著下调(P〈0.01)。结论人参皂苷Rgl能缓解D-半乳糖致衰大鼠脾损伤,其机制可能与抑制氧化损伤和下调P53-P21-RB信号通路有关。
Objective To investigate the effect of ginsenoside Rgl on the spleen structure and function of aging rats and its relative mechanism. Methods Forty SD rats were randomly divided into normal control group, aging model group ( D-galactose 120 mg/kg, qd× 42 d) , Rgl intervention group ( D-galaetose 120 mg/kg, qd × 42 d and Rgl 20 mg/kg, from day 15th,qd×28 d) and Rgl control group. After finishing injections the spleen index was meas- ured, paraffin sections were then made to observe spleen microscopic structure. Senescence-associated β-Galactosi- dase(SA-β-Gal) stain was used to detect aging splenocytes. The proliferative capacity of splenocytes stimulated with Concanavalin A (ConA) was measured by CCK-8. The content of IL-2, IL-6 and advanced glycosylation end products(AGEs) was detected by ELISA. The level of ROS was analyzed by flow cytometry(FCM). Malondialde- hyde ( MDA), superoxide dismutase (SOD) were detected by enzymatic assay. The expression of seneseence-assoei-ated protein P53 ,P21 and RB were detected by Western blot analysis. Results Comparing the Rgl intervention group with the aging model group, spleen index, splenic white pulp area proportion, the proliferative capacity of splenocytes were significantly increased (P 〈0. 05) ;The secretory capability of IL-2 and IL-6, the active content of SOD were obviously increased(P 〈0. 01 ) ;The percentage of SA-β-Gal positive splenocytes, the productions of ROS and MDA were significantly decreased (P 〈0.01 ) ;The production of AGEs was decreased (P 〈0. 05 ) ;The expressions of P53 ,P21 and Rb were also significantly down-regulated (P 〈0. 01 ). Conclusions Ginsenoside Rgl relieves injure of the spleen in aging rats induced by D-galactose. It is suggested that the mechanism may be Rgl in- hibiting oxidative stress and down-regulating P53-P21-RB signaling pathway.