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目的探讨加味四君子汤对泼尼松干预的阿霉素肾病大鼠骨代谢的影响。方法制作大鼠阿霉素肾病模型,将50只SD大鼠随机分为5组,即模型组、激素组、中药组、中药加激素组及正常组。各组于造模后7、21、35天收集24 h尿标本,以双缩脲比色法测定大鼠24 h尿蛋白,以ELISA法测定各组血清骨保护素(osteoprotegerin,OPG)、核因子κB受体活化因子配体(receptor activator for NF-κB ligand,RANKL)、骨钙素(osteocalcin,BGP)以及抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRACP)水平,采用实时定量PCR和Western blot检测胫骨组织中OPG、RANKL的表达。结果 (1)与正常组比较,模型组第7、21、35天24 h尿蛋白均有不同程度增高(P<0.05,P<0.01);与模型组同期比较,激素组及中药加激素组24 h尿蛋白均有不同程度下降,差异有统计学意义(P<0.05,P<0.01);并随着治疗时间的延长,降低更明显(P<0.05,P<0.01);中药组治疗35天,与模型组比较,差异亦有统计学意义(P<0.05)。(2)与本组21天比较,激素组TRACP、RANKL均明显升高(P<0.05,P<0.01);与模型组比较,第21、35天,激素组TRACP、RANKL均升高(P<0.05,P<0.01),OPG、BGP均降低(P<0.05,P<0.01);与中药组同期比较,激素组、中药加激素组OPG均降低,RANKL均升高(P<0.05,P<0.01);且激素组TRACP升高,BGP降低(P<0.05,P<0.01);与激素组同期比较,中药加激素组OPG、BGP升高(P<0.05,P<0.01),RANKL降低(P<0.01);第35天,TRACP降低(P<0.01)。(3)与正常组比较,模型组第21天OPG、RANKL mRNA升高(P<0.05,P<0.01),第35天OPG、RANKL mRNA降低(P<0.01)。与中药组同期比较,激素组OPG mRNA降低(P<0.01),RANKL mRNA升高(P<0.05);中药加激素组OPG mRNA降低(P<0.05)。(4)与本组21天比较,激素组OPG降低(P<0.05),RANKL升高(P<0.05),中药加激素组RANKL降低(P<0.05)。与模型组同期比较,激素组OPG降低(P<0.01),RANKL升高(P<0.01)。与中药组同期比较,激素组、中药加激素组OPG均降低(P<0.01),RANKL均升高(P<0.01);与激素组同期比较,中药加激素组OPG均升高(P<0.01),RANKL均降低(P<0.01)。结论泼尼松能通过OPG/RANKL/RANK通路诱导骨质疏松。加味四君子汤在降蛋白尿的同时,可通过上述通路促进成骨细胞分化,抑制破骨细胞生成,从而减缓泼尼松诱导的骨质疏松的形成。
Objective To investigate the effect of Jiawei Sijunzi Decoction on bone metabolism in adriamycin-induced nephropathy rats treated with prednisone. Methods Adriamycin nephropathy model was made in rats. Fifty SD rats were randomly divided into five groups: model group, hormone group, TCM group, TCM plus hormone group and normal group. Twenty-four hours urine samples were collected at 7,21,35 days after modeling. Urine protein was measured by biuret colorimetric method. Serum levels of osteoprotegerin (OPG) and nuclear (RANKL), osteocalcin (BGP) and tartrate-resistant acid phosphatase (TRACP) were detected by real-time quantitative PCR Western blot was used to detect the expression of OPG and RANKL in tibia. Results (1) Compared with the normal group, urinary protein in 24 h of model group was increased to different extents (P <0.05, P <0.01) on the 7th, 21st and 35th day. Compared with the model group, (P <0.05, P <0.01). With the prolongation of treatment time, the decrease was more obvious (P <0.05, P <0.01) Day, compared with the model group, the difference was also statistically significant (P <0.05). (2) The levels of TRACP and RANKL in hormone group were significantly higher than those in the 21-day group (P <0.05, P <0.01). Compared with the model group, on the 21st and 35th day, (P <0.05, P <0.01, P <0.01, P <0.01, P <0.01, P <0.01). Compared with the same period of the traditional Chinese medicine group, the OPG and the RANKL of the hormone group and the traditional Chinese medicine plus hormone group were all decreased (P <0.05, P <0.01). Compared with the hormone group, the levels of OPG and BGP increased (P <0.05, P <0.01) and the levels of RANKL decreased (P <0.01). On the 35th day, TRACP decreased (P <0.01). (3) Compared with the normal group, OPG and RANKL mRNA increased (P <0.05, P <0.01) on the 21st day in model group and OPG and RANKL mRNA decreased on the 35th day (P <0.01). Compared with the same period of the traditional Chinese medicine group, OPG mRNA in the hormone group decreased (P <0.01) and RANKL mRNA increased (P <0.05). The OPG mRNA in the hormone group decreased (P <0.05). (4) Compared with the 21-day group, OPG decreased (P <0.05) and RANKL increased (P <0.05) in the hormone group and decreased RANKL (P <0.05). Compared with the model group, OPG in the hormone group decreased (P <0.01) and RANKL increased (P <0.01). Compared with the same period of the traditional Chinese medicine group, the OPG of the hormone group and the traditional Chinese medicine plus hormone group decreased (P <0.01) and the expression of RANKL increased (P <0.01) ), RANKL decreased (P <0.01). Conclusion Prednisone can induce osteoporosis through the OPG / RANKL / RANK pathway. Modified Sijunzi Decoction in reducing proteinuria at the same time, through the above pathway to promote osteoblast differentiation, inhibition of osteoclastogenesis, thereby reducing the prednisone-induced osteoporosis formation.