复方参鹿颗粒含药血清对较低危骨髓增生异常综合征p38MAPK磷酸化的抑制作用  

Inhibiting Effects of Compound Shenlu Granules Containing Serum on p38MAPK Phosphorylation of Lower-risk MDS

作　　者：张文君[1] 赵琳[2] 赵艺涵 徐培[2] 王子元[3] ZHANG Wen-jun;ZHAO Lin;ZHAO Yi-han;XU Pei;WANG Zi-yuan(Department of Hematology,Tongji Hospital of Tongji University,Shanghai (200092);Department of Hematology,Shuguang Hospital Affiliated to Shanghai University of TCM,Shanghai (200021);Department of Pathology,Shuguang Hospital Affiliated to Shanghai University of TCM,Shanghai (200021))

机构地区：[1]同济大学附属同济医院血液科,上海200092 [2]上海中医药大学附属曙光医院血液科,上海200021 [3]上海中医药大学附属曙光医院病理科,上海200021

出　　处：《中国中西医结合杂志》2019年第1期82-87,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金青年科学基金项目(No.81403233)

摘　　要：目的观察复方参鹿颗粒(CSG)含药血清对较低危骨髓增生异常综合征(lower-risk MDS)骨髓单个核细胞(BMMNC)p38MAPK磷酸化、CD34^+细胞凋亡和正常造血干/祖细胞集落(CFU-HSPC)形成的影响。方法 16只清洁级Wistar大鼠,随机分为空白血清组、CSG组,每组8只,制备空白血清和CSG含药血清。体外骨髓细胞培养分为空白血清组、CSG组、SB203580组和SB202474组。分离lower-risk MDS患者BMMNC,与10%各组血清或p38抑制剂共培养,Annexin V/PI双染流式细胞术检测CD34^+细胞凋亡率, Western Blot检测p38、p-p38蛋白表达。MACS分离骨髓CD34^+细胞,用半固体甲基纤维素与10%各组血清或p38抑制剂共培养,观察CFU-HSPC形成,FISH检测集落中异常克隆细胞群体的比例。结果与空白血清和SB202474组比较,CSG、SB203580组的凋亡率、p-p38蛋白表达减少(P<0.05);CFU-HSPC形成数增加(P<0.01);形成的集落中异常克隆细胞比例降低(P<0.05)。CSG和SB203580组的p-p38蛋白表达和异常克隆细胞比例呈正相关(P<0.01)。结论 CSG含药血清能改善lower-risk MDS骨髓无效造血,其机制可能与抑制p38蛋白磷酸化,减少骨髓CD34^+细胞凋亡,并恢复骨髓正常造血细胞的优势生长有关。Objective To explore the effects of Compound Shenlu Granules(CSG) containing serum on the phosphorylation of p38MAPK, the apoptosis of CD34+ cells of lower-risk myelodysplastic syndromes(MDS) bone marrow and the normal colony forming unit of hematopoietic stem/progenitor cell(CFU-HSPC) formation. Methods Sixteen clean grade Wistar mice were randomly divided into blank serum group and CSG group, 8 in each group. The blank serum and CSG containing serum were prepared. Bone marrow cells cultured in vitro were divided into 4 groups: blank serum group, CSG group, SB203580 group, SB202474 group. Bone marrow mononuclear cells were isolated from patients with lower-risk MDS, then cultured with 10% serums of different groups or p38 inhibitor in vitro. The apoptosis rates of bone marrow CD34+ cells were detected by annexin V/PI double staining flow cytometry. The expressions of p38 and p-p38 protein were tested using Western Blot. Bone marrow CD34+ cells were isolated by MACS and cultured with 10% serums of different groups or p38 inhibitors. Formation of CFU-HSPC was observed. The percentages of aberrant clones in CFU-HSPC were detected by FISH. Results Compared with the blank serum group and SB202474 group, the apoptosis rates, the expressions of p-p38 protein of CSG and SB203580 group were lower(P<0.05), the numbers of CFU-HSPC in CSG group were higher(P<0.01), the percentages of aberrant clones in CFU-HSPC were lower(P<0.05). P-p38 protein expressions and the proportions of abnormal clone cells of CSG and SB203580 group were positively correlated(P<0.01). Conclusions CSG-containing serum could improve the ineffective hematopoiesis of lower-risk MDS patients. The mechanism may be related to the inhibition of p38 protein phosphorylation, the reduction of apoptosis in bone marrow CD34+ cells and the restoration of normal hematopoietic in bone marrow.

关 键 词：复方参鹿颗粒 含药血清 骨髓增生异常综合征 P38丝裂原活化蛋白激酶 

分 类 号：R551.3[医药卫生—血液循环系统疾病]