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210.6 Rnt/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0.4,0.08,0.016 g/L鸡血藤活性成分SS8,1次/d,连续给药21 d.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3 d,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=1.847 7,P=0.114 1;t=1.473 1,P=0.191 1;t=1.473 1,P=0.1911];给药后21 d,SS8低剂量组与对照组基本持平[(43.60±2.07),(47.00±3.92)个;t=1.534 0,P=0.175 9],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=16.889 6,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3 d,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7 d,各组均开始恢复;给药后21 d,SS8低剂量组与对照组基本相似[(44.50±1.29),(42.67±7.23)个;t=0.511 9,P=0.630 5],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=5.355 3,P＜0.05;t=12.822 3,P＜0.05].③各组爆式红系造血祖细胞集落生长情况:给药后3 d,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21 d,SS8低剂量组仍与对照组相似[(5.00±1.82),(3.00±0.82)个;t=2.003 8,P=0.091 9],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=9.311 9,P＜0.05;t=13.973 5,P＜0.01].④各组巨核系造血祖细胞集落生长情况:给药后3 d,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=4.941 2,P=0.007 8);给药后21 d,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=3.851 2,P=0.008 4;t=0.975 3.P=0.361 7;t=1.283 7,P=0.246 6].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.
揹景:機體造血調控的關鍵在于造血榦細胞和造血祖細胞,被抑製的造血榦/祖細胞的增殖、分化、成熟和釋放中的任何一箇環節的增彊,都可以促進機體造血功能的恢複.目的:分析鷄血籐活性成分SS8對骨髓抑製小鼠粒單繫造血祖細胞、紅繫造血祖細胞、巨覈繫造血祖細胞增殖的影響.設計:隨機對照實驗.單位:中國人民解放軍總醫院臨床藥理研究室.材料:實驗于2002-02/2002-06在解放軍總醫院藥理研究室完成.選取健康昆明種小鼠20隻,隨機分為正常組、對照組、SS8高劑量組、SS8中劑量組、SS8低劑量,4隻/組.方法:除正常組外,其餘各組小鼠均以60Coγ射線全身亞緻死量輻射(照射率210.6 Rnt/min,照射劑量400cGy/min,照射時間110.7s),于照射後噹天正常組和對照組腹腔註射無菌註射用水,SS8高、中、低劑量組分彆腹腔註射0.4,0.08,0.016 g/L鷄血籐活性成分SS8,1次/d,連續給藥21 d.給藥後第3,7,14,21天時分彆行頸椎脫位法處死小鼠,收集股骨骨髓造血祖細胞,採用甲基纖維素半固體培養法,對長期接受SS8治療後的骨髓抑製小鼠的造血祖細胞進行體外培養.主要觀察指標:各組粒單繫造血祖細胞、紅繫造血祖細胞、各組爆式紅繫造血祖細胞、各組爆式紅繫造血祖細胞的集落生長情況.結果:實驗納入20隻小鼠全部進入結果分析.①各組粒單繫造血祖細胞集落生長情況:給藥後3 d,正常組粒單繫造血祖細胞集落數為(180.67±5.03)箇,SS8高、中、低劑量組與對照組基本相似[(0.75±0.96),(1.75±0.50),(1.75±0.96),(1.75±0.96)箇;t=1.847 7,P=0.114 1;t=1.473 1,P=0.191 1;t=1.473 1,P=0.1911];給藥後21 d,SS8低劑量組與對照組基本持平[(43.60±2.07),(47.00±3.92)箇;t=1.534 0,P=0.175 9],SS8中、高劑量組均明顯高于對照組[(90.60±3.36),(93.00±3.16),(47.00±3.92)箇;t=16.889 6,P＜0.05;t=18.2718,P＜0.05].②各組紅繫造血祖細胞集落生長情況:給藥後3 d,對照組和SS8高、中、低劑量組與正常組比較,紅繫造血祖細胞數目明顯降低;至給藥後7 d,各組均開始恢複;給藥後21 d,SS8低劑量組與對照組基本相似[(44.50±1.29),(42.67±7.23)箇;t=0.511 9,P=0.630 5],SS8中、高劑量組均明顯高于對照組[(62.50±2.08),(93.25±3.10),(42.67±7.23)箇,t=5.355 3,P＜0.05;t=12.822 3,P＜0.05].③各組爆式紅繫造血祖細胞集落生長情況:給藥後3 d,對照組和SS8高、中、低劑量組爆式紅繫造血祖細胞均降至最低,分彆為(1.00±1.00),(7.00±1.00),(6.00±2.00),(6.00±2.00)箇,但SS8高、中、低劑量組仍高于對照組(P均＜0.05);給藥後21 d,SS8低劑量組仍與對照組相似[(5.00±1.82),(3.00±0.82)箇;t=2.003 8,P=0.091 9],SS8中、高劑量組均明顯高于對照組[(15.25±2.50),(16.25±1.71),(3.00±0.82)箇;t=9.311 9,P＜0.05;t=13.973 5,P＜0.01].④各組巨覈繫造血祖細胞集落生長情況:給藥後3 d,由于照射後小鼠骨髓造血祖細胞的增殖能力明顯受損,SS8高、中、低劑量組及對照組巨覈繫造血祖細胞數目較正常組顯著降低,但SS8高、中、低劑量組仍高于對照組[(2.67±0.58),(1.33±0.58),(1.33±0.58),(0.33±0.58)箇],且SS8高劑量組與對照組比較有顯著性差異(t=4.941 2,P=0.007 8);給藥後21 d,SS8高、中、低劑量組均明顯高于對照組[(2.50±0.58),(2.00±0.32),(1.50±0.58),(1.00±0.52)箇;t=3.851 2,P=0.008 4;t=0.975 3.P=0.361 7;t=1.283 7,P=0.246 6].結論:鷄血籐活性成分SS8低劑量對骨髓抑製小鼠造血祖細胞的增殖無顯著刺激作用,而中、高劑量均可顯著升高骨髓抑製後粒單繫造血祖細胞、紅繫造血祖細胞、巨覈繫造血祖細胞集落產率,且隨時間的延長刺激作用逐漸加彊,且高劑量的刺激作用明顯優于中劑量.提示造血祖細胞的增生是機體造血的重要環節,鷄血籐活性成分SS8對骨髓抑製小鼠造血祖細胞的增殖有明顯刺激作用.
배경:궤체조혈조공적관건재우조혈간세포화조혈조세포,피억제적조혈간/조세포적증식、분화、성숙화석방중적임하일개배절적증강,도가이촉진궤체조혈공능적회복.목적:분석계혈등활성성분SS8대골수억제소서립단계조혈조세포、홍계조혈조세포、거핵계조혈조세포증식적영향.설계:수궤대조실험.단위:중국인민해방군총의원림상약리연구실.재료:실험우2002-02/2002-06재해방군총의원약리연구실완성.선취건강곤명충소서20지,수궤분위정상조、대조조、SS8고제량조、SS8중제량조、SS8저제량,4지/조.방법:제정상조외,기여각조소서균이60Coγ사선전신아치사량복사(조사솔210.6 Rnt/min,조사제량400cGy/min,조사시간110.7s),우조사후당천정상조화대조조복강주사무균주사용수,SS8고、중、저제량조분별복강주사0.4,0.08,0.016 g/L계혈등활성성분SS8,1차/d,련속급약21 d.급약후제3,7,14,21천시분별행경추탈위법처사소서,수집고골골수조혈조세포,채용갑기섬유소반고체배양법,대장기접수SS8치료후적골수억제소서적조혈조세포진행체외배양.주요관찰지표:각조립단계조혈조세포、홍계조혈조세포、각조폭식홍계조혈조세포、각조폭식홍계조혈조세포적집락생장정황.결과:실험납입20지소서전부진입결과분석.①각조립단계조혈조세포집락생장정황:급약후3 d,정상조립단계조혈조세포집락수위(180.67±5.03)개,SS8고、중、저제량조여대조조기본상사[(0.75±0.96),(1.75±0.50),(1.75±0.96),(1.75±0.96)개;t=1.847 7,P=0.114 1;t=1.473 1,P=0.191 1;t=1.473 1,P=0.1911];급약후21 d,SS8저제량조여대조조기본지평[(43.60±2.07),(47.00±3.92)개;t=1.534 0,P=0.175 9],SS8중、고제량조균명현고우대조조[(90.60±3.36),(93.00±3.16),(47.00±3.92)개;t=16.889 6,P＜0.05;t=18.2718,P＜0.05].②각조홍계조혈조세포집락생장정황:급약후3 d,대조조화SS8고、중、저제량조여정상조비교,홍계조혈조세포수목명현강저;지급약후7 d,각조균개시회복;급약후21 d,SS8저제량조여대조조기본상사[(44.50±1.29),(42.67±7.23)개;t=0.511 9,P=0.630 5],SS8중、고제량조균명현고우대조조[(62.50±2.08),(93.25±3.10),(42.67±7.23)개,t=5.355 3,P＜0.05;t=12.822 3,P＜0.05].③각조폭식홍계조혈조세포집락생장정황:급약후3 d,대조조화SS8고、중、저제량조폭식홍계조혈조세포균강지최저,분별위(1.00±1.00),(7.00±1.00),(6.00±2.00),(6.00±2.00)개,단SS8고、중、저제량조잉고우대조조(P균＜0.05);급약후21 d,SS8저제량조잉여대조조상사[(5.00±1.82),(3.00±0.82)개;t=2.003 8,P=0.091 9],SS8중、고제량조균명현고우대조조[(15.25±2.50),(16.25±1.71),(3.00±0.82)개;t=9.311 9,P＜0.05;t=13.973 5,P＜0.01].④각조거핵계조혈조세포집락생장정황:급약후3 d,유우조사후소서골수조혈조세포적증식능력명현수손,SS8고、중、저제량조급대조조거핵계조혈조세포수목교정상조현저강저,단SS8고、중、저제량조잉고우대조조[(2.67±0.58),(1.33±0.58),(1.33±0.58),(0.33±0.58)개],차SS8고제량조여대조조비교유현저성차이(t=4.941 2,P=0.007 8);급약후21 d,SS8고、중、저제량조균명현고우대조조[(2.50±0.58),(2.00±0.32),(1.50±0.58),(1.00±0.52)개;t=3.851 2,P=0.008 4;t=0.975 3.P=0.361 7;t=1.283 7,P=0.246 6].결론:계혈등활성성분SS8저제량대골수억제소서조혈조세포적증식무현저자격작용,이중、고제량균가현저승고골수억제후립단계조혈조세포、홍계조혈조세포、거핵계조혈조세포집락산솔,차수시간적연장자격작용축점가강,차고제량적자격작용명현우우중제량.제시조혈조세포적증생시궤체조혈적중요배절,계혈등활성성분SS8대골수억제소서조혈조세포적증식유명현자격작용.
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 suberectus Dunn, on the proliferation of granulocyte-macrophage colony-forming unit (CFU-GM), erythrocyte 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, SS8medium-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 sublethal dose of irradiation by 60Co γ -ray (210.6rontgen/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 (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 SS8low-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,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 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].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 marrow mice.
