鸡血藤活性成分SS8对骨髓抑制小鼠造血祖细胞增殖的作用(英文)

Effect of SS8, an active component extracted from Spatholobus suberectus Dunn,on the proliferation of hemopoietic progenitor cells in marrow-depressed mice

背景:机体造血调控的关键在于造血干细胞和造血祖细胞,被抑制的造血干/祖细胞的增殖、分化、成熟和释放中的任何一个环节的增强,都可以促进机体造血功能的恢复。目的:分析鸡血藤活性成分SS8对骨髓抑制小鼠粒单系造血祖细胞、红系造血祖细胞、巨核系造血祖细胞增殖的影响。设计:随机对照实验。单位:中国人民解放军总医院临床药理研究室。材料:实验于2002-02/2002-06在解放军总医院药理研究室完成。选取健康昆明种小鼠20只,随机分为正常组、对照组、SS8高剂量组、SS8中剂量组、SS8低剂量,4只/组。方法:除正常组外,其余各组小鼠均以60Coγ射线全身亚致死量辐射(照射率210.6Rnt/min,照射剂量400cGy/min,照射时间110.7s),于照射后当天正常组和对照组腹腔注射无菌注射用水,SS8高、中、低剂量组分别腹腔注射0.4,0.08,0.016g/L鸡血藤活性成分SS8,1次/d,连续给药21d。给药后第3,7,14,21天时分别行颈椎脱位法处死小鼠,收集股骨骨髓造血祖细胞,采用甲基纤维素半固体培养法,对长期接受SS8治疗后的骨髓抑制小鼠的造血祖细胞进行体外培养。主要观察指标:各组粒单系造血祖细胞、红系造血祖细胞、各组爆式红系造血祖细胞、各组爆式红系造血祖细胞的集落生长情况。结果:实验纳入20只小鼠全部进入结果分析。①各组粒单系造血祖细胞集落生长情况:给药后3d,正常组粒单系造血祖细胞集落数为(180.67±5.03)个,SS8高、中、低剂量组与对照组基本相似[(0.75±0.96),(1.75±0.50),(1.75±0.96),(1.75±0.96)个;t=1.8477,P=0.1141;t=1.4731,P=0.1911;t=1.4731,P=0.1911];给药后21d,SS8低剂量组与对照组基本持平[(43.60±2.07),(47.00±3.92)个;t=1.5340,P=0.1759],SS8中、高剂量组均明显高于对照组[(90.60±3.36),(93.00±3.16),(47.00±3.92)个;t=16.8896,P<0.05;t=18.2718,P<0.05]。②各组红系造血祖细胞集落生长情况:给药后3d,对照组和SS8高、中、低剂量组与正常组比较,红系造血祖细胞数目明显降低;至给药后7d,各组均开始恢复;给药后21d,SS8低剂量组与对照组基本相似[(44.50±1.29),(42.67±7.23)个;t=0.5119,P=0.6305],SS8中、高剂量组均明显高于对照组[(62.50±2.08),(93.25±3.10),(42.67±7.23)个,t=5.3553,P<0.05;t=12.8223,P<0.05]。③各组爆式红系造血祖细胞集落生长情况:给药后3d,对照组和SS8高、中、低剂量组爆式红系造血祖细胞均降至最低,分别为(1.00±1.00),(7.00±1.00),(6.00±2.00),(6.00±2.00)个,但SS8高、中、低剂量组仍高于对照组(P均<0.05);给药后21d,SS8低剂量组仍与对照组相似[(5.00±1.82),(3.00±0.82)个;t=2.0038,P=0.0919],SS8中、高剂量组均明显高于对照组[(15.25±2.50),(16.25±1.71),(3.00±0.82)个;t=9.3119,P<0.05;t=13.9735,P<0.01]。④各组巨核系造血祖细胞集落生长情况:给药后3d,由于照射后小鼠骨髓造血祖细胞的增殖能力明显受损,SS8高、中、低剂量组及对照组巨核系造血祖细胞数目较正常组显著降低,但SS8高、中、低剂量组仍高于对照组[(2.67±0.58),(1.33±0.58),(1.33±0.58),(0.33±0.58)个],且SS8高剂量组与对照组比较有显著性差异(t=4.9412,P=0.0078);给药后21d,SS8高、中、低剂量组均明显高于对照组[(2.50±0.58),(2.00±0.32),(1.50±0.58),(1.00±0.52)个;t=3.8512,P=0.0084;t=0.9753,P=0.3617;t=1.2837,P=0.2466]。结论:鸡血藤活性成分SS8低剂量对骨髓抑制小鼠造血祖细胞的增殖无显著刺激作用,而中、高剂量均可显著升高骨髓抑制后粒单系造血祖细胞、红系造血祖细胞、巨核系造血祖细胞集落产率,且随时间的延长刺激作用逐渐加强,且高剂量的刺激作用明显优于中剂量。提示造血祖细胞的增生是机体造血的重要环节,鸡血藤活性成分SS8对骨髓抑制小鼠造血祖细胞的增殖有明显刺激作用。

BACKGROUND： Hematopoietic stem cells （HSC） and hematopoietic progenitor cells （HPC） are the key to hematopoietic regulation. The enhanced effect on any link in proliferation, differentiation, maturation and release of the depressed HSC/HPC can accelerate the recovery of hematopoiesis. OBJECTIVE： To investigate the effect of SS8, an active component extracted from Spatholobus subereetus Dunn, on the proliferation of granulocyte-macrophage colony-forming unit （CFU-GM）, erythroeyte colonyforming unit （CFU-E）, and megakaryocyte colony-forming unit （CFU-Meg） in bone marrow-depressed mice. DESIGN： Randomized controlled trial. SETTING： Department of Clinical Pharmacology, General Hospital of Chinese PLA. MATERIALS： The experiment was conducted in the Department of Clinical Pharmacology, General Hospital of Chinese PLA, from February to June 2002. Totally 20 Kunming healthy mice were recruited and randomized into normal group, control group, SS8 high-dose group, SS8 medium-dose group, and SS8 low-dose group with 4 mice in each group. INTERVENTIONS： All the mice except the mice in normal group had been given total body suhlethal dose of irradiation by ^60Co γ-ray （210.6 rontgen/min, 400cGy/min dose rate, irradiation time of 110.7 s. Normal saline was injected intraperitoneally into the mice in normal group and control group on the day of irradiation. Stored SS8 solution 0.4, 0.08, and 0.016 g/L was intraperitoneally injected into the mice in SS8 high-, medium- and low-dose groups, respectively, once a day, immediately after irradiation for 21 consecutive days. By the end of the third, seventh, fourteenth and twenty-first day after injection, the mice were killed by dislocating cervical vertebra, and femoral marrow cells of them were collected for cultivation with methylcellulose demisolid method. HPC of depressed bone marrow mice was cultured in vitro after treatment of SS8. MAIN OUTCOME MEASURES： CFU-GM, CFU-E, and CFU-Meg in each group. RESULTS： Totally 20 mice included in the experimental group all entered the result analysis. ①Growth of CFU-GM in each group： it was （180.67±5.03）in normal group 3 days after administration, and it was basically the same in SS8 high-, medium-and low-dose groups [（0.75±0.96）, （1.75±0.50）, （1.75±0.96）,（1.75±0.96; t=1.8477, = 0.114 1; t=1.473 1, P=0.191 1; t=1.473 1, P=0.191 1] as in control group. The number of SS8 low-dose group had the same level as that in control group 21 days after administration [（43.60±2.07）,（47.00±3.92） ;t=1.534 0,P=0.175 9], and it was obviously higher in SS8 medium-and high-dose groups than that in control group [ （90.60±3.36）, （93.00±3.16 ）, （47.00±3.92） ; t = 16.889 6, P 〈 0.05;t=18.271 8,P 〈 0.05]. ②Growth of CFU-E in each group： The number of CFU-E was obviously decreased when the number was compared between control group and different SS8 dose groups and normal group 3 days after administration. The number in each group began to recover 7 days after administration; the number in SS8 low-dose group was similar to that in control group 21 days after administration [（44.50±1.29）, （42.67±7.23） ;t=0.511 9,P=0.630 5], it was obviously higher in SS8 medium-and high-dose groups than in control group[（62.50±2.08）, （93.25±3.10）, （42.67±7.23）,t=5.355 3,P〈0.05;t =12.822 3,/9 〈 0.05].③The number of BFU-E in control group and SS8 high-, medium-, and low-dose groups was decreased to the lowest level 3 days after administration [（1.00±1.00）, （7.00±1.00）, （6.00±2.00）, （6.00±2.00）, respectively], but-the number in the administration group was still higher than that in control group （all P 〈 0.05）. The number in SS8 low-dose group was similar to that in control group [（5.00±1.82）, （3.00±0.82） ;t=2.003 8,P=0.091 9], and that in SS8 medium-and highdose groups was obviously than that in control group [（15.25±2.50）, （ 16.25±1.71 ）, （3.00±0.82） ; t=9.3119 ,P〈 0.05 ;t=13.973 5 ,P 〈 0.01]. ④Tbe growth of CFU-Meg in each group： Because the proliferation capability of bone marrow HPC of mice was obviously injured, the number of CFU-Meg in SS8 high-, medium-and low-dose groups and control group was significantly decreased compared to that in control group, but it was greater in the three SS8 dose groups than in control group [（2.67±0.58）, （1.33±0.58）, （1.33±0.58）, （0.33±0.58）]. There was significant difference between SS8 high-dose group and control group （t =4.941 2,P=0.007 8）.The number of CFU-Meg in SS8 low-and medium-dose groups was decreased to the level of control group 7 days after administration, （0.75±0.36）in control group, （1.50±0.18）in medium-dose group, and （1.25±0.96）in low-dose group 14 days after administration. It was （1.00±0.52）in control group, （2.00±0.32）in medium-dose group, and （1.50±0.58）in low-dose group 21 days after administration, which was significantly higher than that in control group （t =3.726 8,P=0.009 8;t=0.975 3,P=0.361 7;t=3.275 6,P=0.016 9; t= 1.283 7,P=0.246 6）. CONCLUSION： SS8, an active component from Spatholobus suberectus Dunn, at low dose, has no obvious stimulating effect on the proliferation of HPC in depressed bone marrow mice. After high-dose and mediumdose SS8 solution was intraperitoneally injected into the mice, there was a significant augmenting effect on the proliferation of CFU-GM, CFU-E, BFU-E and CFU-Meg, and the augmenting effect of SS8 enhanced as time prolonged and the effect of high-dose SS8 solution was superior to that of medium-dose SS8 solution, suggesting that the proliferation of HPC is the key link of hematopoietic regulation. Therefore, SS8 has obvious stimulating effect on the proliferation of HPC in depressed bone