木豆叶提取物对股骨头坏死大鼠血液流变学及骨髓间充质干细胞分化的影响

Effects of folium cajani on hemorheology and BMSCs differentiation in rats with femoral head necrosis

目的观察木豆叶提取物对股骨头坏死大鼠血液流变学及骨髓间充质干细胞(BMSCs)分化的影响。方法2017年10月—2018年12月于湖北省十堰市太和医院生物医学研究所进行实验,将50只SPF级雄性SD大鼠按随机数字表法均分为对照组、模型组、低剂量组、中剂量组、高剂量组,各10只,后4组大鼠采用大肠杆菌脂多糖(40μg/kg)与甲基强的松龙(20 mg/kg)建立激素性股骨头坏死大鼠模型;而后低剂量组、中剂量组、高剂量组分别采用不同剂量木豆叶提取物(0.48 g/kg、0.96 g/kg、1.44 g/kg)连续干预4周。观察各组大鼠血液流变学、凝血功能及坏死股骨修复情况;分离各组股骨头坏死大鼠BMSCs,采用不同浓度含木豆叶血清处理BMSCs 1周,观察三酰甘油(TG)、碱性磷酸酶(ALP)、骨钙素(BGP)活性、钙化结节面积。结果与对照组比较,模型组大鼠全血低切黏度、全血中切黏度、全血高切黏度、血浆黏度均升高(P<0.01),凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)、凝血酶时间(TT)均降低(P<0.01);与模型组比较,低剂量组、中剂量组、高剂量组大鼠全血低切黏度、全血中切黏度、全血高切黏度、血浆黏度均降低,PT、APTT、TT均升高(P均<0.05)。对照组股骨头软骨细胞排列有序且骨小梁完整,模型组股骨头软骨细胞排列紊乱、骨小梁变窄、坏死细胞增多,木豆叶低剂量组、中剂量组、高剂量组股骨头软骨细胞损伤程度较模型组明显减轻。体外BMSCs实验结果显示,与对照组比较,低剂量亚组、中剂量亚组、高剂量亚组BMSCs中TG水平降低(F/P=58.108/0.000),ALP、BGP活性与钙化结节面积增加(F/P=20.140/0.000、18.914/0.000、19.659/0.000)。结论木豆叶提取物对激素性股骨头坏死大鼠血液流变学、股骨头坏死修复有一定的改善作用,可能与调节血液黏度、凝血功能及BMSCs向成骨分化能力有关。

Objective To observe the effects of folium cajani(FC)on hemorheology and BMSCs differentiation in rats with femoral head necrosis.Methods During the period from October 2017 to December 2018 at Biomedicine Institute of Taihe Hospital,Hubei Shiyan,50 SPF male SD rats were randomly averagely divided into control group,model group,low-dose group,middle-dose group and high-dose group.Except control group,scherichia coli lipopolysaccharide(40μg/kg)and methylprednisolone(20 mg/kg)were applied to establish rat models of hormonal femoral head necrosis.The low-dose,middle-dose and high-dose groups were continuously intervened with FC extract of different doses(0.48 g/kg,0.96 g/kg,1.44 g/kg)for 4 weeks.The hemorheology,coagulation function and necrotic femoral repair in each group were observed.The rat models of femoral head necrosis were prepared by the above method.Bone marrow mesenchymal stem cells(BMSCs)in rats with femoral head necrosis in each group were isolated.BMSCs were treated with serum of different concentrations which contained FC for 1 week.The triglyceride(TG),alkaline phosphatase(ALP),osteocalcin(BGP)activity and calcified nodule area were observed.Results Compared with control group,whole blood low-shear,middle-shear and high-shear viscosity,and plasma viscosity were increased in model group,low-dose group,middle-dose group and high-dose group(P<0.01),while prothrombin time(PT),activated partial thromboplastin time(APTT)and thrombin time(TT)were all decreased(P<0.01).Compared with model group,whole blood low-shear,middle-shear and high-shear viscosity,and plasma viscosity were decreased in low-dose group,middle-dose group and high-dose group,while PT,APTT and TT were all increased(P<0.05).The femoral head chondrocytes in control group were arranged in order,and trabecular bone was complete.The arrangement of femoral head chondrocytes in model group was disordered,trabecular bone was narrowed,necrotic cells was increased.The femoral head chondrocyte damage in low-dose,middle-dose and high-dose group was significantly milder than that in model group.The results of BMSCs in vitro test showed that compared with blank group,TG content of BMSCs was decreased in low-dose,middle-dose and high-dose subgroups(F/P=58.108/0.000),while ALP,BGP activity and calcified nodule area were increased(F/P=20.140/0.000,18.914/0.000,19.659/0.000).Conclusion There are certain improvement effects of FC on hemorheology and femoral head necrosis repair in rats with hormonal femoral head necrosis,which may be related to regulating blood viscosity,coagulation function and BMSCs osteogenic differentiation ability.