健脾补肾方调控p53/p21细胞衰老通路抑制辐射小鼠造血系统损伤的机制研究OA
Mechanism of Jianpi Bushen Prescription in Inhibiting Hematopoietic System Damage via the p53/p21 Cell Senescence Pathway in Radiation-Exposed Mice
目的 观察健脾补肾方对辐射损伤模型小鼠p53/p21 细胞衰老信号通路的调控作用,探讨健脾补肾方抑制辐射造血系统损伤的机制.方法 选用 30 只SPF级昆明种小鼠,采用随机数表法选取6 只小鼠作为正常对照组,剩余 24 只小鼠接受 6.0戈瑞(Gy)X射线单次全身照射以建立动物模型,造模成功后将小鼠随机分为模型组、健脾补肾方小剂量组、健脾补肾方大剂量组、复方阿胶浆组.药物灌胃处理后第14 天,收集小鼠外周血,分离小鼠股骨,收集骨髓.血细胞计数仪检测小鼠外周血白细胞、血红蛋白、血小板;苏木精-伊红(HE)染色观察骨髓组织病理学改变;ELISA检测骨髓组织衰老相关β-半乳糖苷酶(SA-β-Gal)浓度;流式细胞术检测骨髓细胞周期;实时荧光定量PCR检测骨髓组织p53、p21、细胞周期蛋白E1(CyclinE1)、细胞周期蛋白依赖性激酶2(CDK2)mRNA表达;Western blot检测骨髓组织p53、磷酸化(p)-p53、p21、CyclinE1、CDK2 蛋白的表达.结果 与正常组比较,模型组小鼠外周血白细胞计数、血红蛋白、血小板计数均显著下降(P<0.01),小鼠骨髓组织结构破坏明显,骨髓细胞大量坏死,骨髓细胞数量显著减少,脂肪细胞显著增多,骨髓组织 SA-β-Gal 浓度显著升高(P<0.01),骨髓 G0/G1 期细胞比例显著升高(P<0.01),S期、G2/M期细胞比例显著下降(P<0.01),骨髓组织 p53、p21 mRNA 及蛋白表达水平均显著升高(P<0.01),CyclinE1、CDK2 mRNA及蛋白表达水平均显著下降(P<0.01).与模型组比较,各药物干预组小鼠上述指标均有明显改善(P<0.05).药物干预组中,健脾补肾方小剂量组疗效优于健脾补肾方大剂量组(P<0.05),与复方阿胶浆组疗效比较差异无统计学意义(P>0.05).结论 健脾补肾方可能通过靶向抑制p53/p21 细胞通路,改善造血细胞衰老状态,促进造血细胞增殖,升高外周血细胞,从而促进造血系统损伤的恢复.
Objective To observe the regulatory effect of Jianpi Bushen prescription on the p53/p21 cell senescence signaling pathway in a radiation-induced mouse model,and to explore its mechanism in inhibiting hematopoietic system damage.Methods Among 30 mice,6 were randomly selected as the normal control group,and the remaining 24 mice received a single whole-body irradiation of 6.0 Gy X-ray to establish the model.After modeling,the mice were randomly divided into model group,low-dose Jianpi Bushen prescription group,high-dose Jianpi Bushen prescription group,and compound E-jiao slurry group.On day 14 after intragastric administration,peripheral blood was collected,femurs were isolated,and bone marrow was harvested.White blood cell(WBC)count,hemoglobin(Hb),and platelet(PLT)levels were measured using a hematology analyzer.Bone marrow histopathological changes were observed by hematoxylin-eosin(HE)staining.Senescence-associated β-galactosidase(SA-β-Gal)Concentration in bone marrow was detected by ELISA.Bone marrow cell cycle was analyzed by flow cytometry.The mRNA expression levels of p53,p21,CyclinE1,and CDK2 in bone marrow tissue were detected by RT-qPCR.The protein expression levels of p53,p-p53,p21,CyclinE1,and CDK2 were detected by Western blot.Results Compared with the normal group,the model group showed significantly decreased WBC count,Hb,and PLT levels(P<0.01),marked destruction of bone marrow tissue structure,extensive necrosis of bone marrow cells,significantly reduced bone marrow cell count,significantly increased adipocytes,significantly elevated SA-β-Gal Concentration(P<0.01),significantly increased proportion of cells in G0/G1 phase(P<0.01),significantly decreased proportions of cells in S and G2/M phases(P<0.01),significantly upregulated mRNA and protein expression of p53 and p21(P<0.01),and significantly downregulated mRNA and protein expression of CyclinE1 and CDK2(P<0.01).Compared with the model group,all drug intervention groups significantly improved the above indicators(P<0.05).Among the drug intervention groups,the low-dose Jianpi Bushen prescription group showed better efficacy than the high-dose group(P<0.05),and no significant difference was observed compared with the compound E-jiao slurry group(P>0.05).Conclusion Jianpi Bushen prescription may promote the recovery of hematopoietic system damage by inhibiting the p53/p21 cell senescence pathway,thereby improving the senescence state of hematopoietic cells,promoting hematopoietic cell proliferation,and increasing peripheral blood cell counts.
肖静静;何东初;张梦云;张勇;丁晓娟;伍伟
中部战区总医院,武汉 430070中部战区总医院,武汉 430070中部战区总医院,武汉 430070中部战区总医院,武汉 430070中部战区总医院,武汉 430070中部战区总医院,武汉 430070
医药卫生
健脾补肾方p53/p21细胞衰老辐射损伤
Jianpi Bushen prescriptionp53/p21Cell senescenceRadiation injury
《中国中医基础医学杂志》 2026 (9)
1939-1946,8
国家自然科学基金面上项目(82574833,81873153,81273905)湖北省中医药管理局中医药重点项目(ZY2023Z010)湖北省自然科学基金青年项目(2024AFB538)
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