左归丸对骨髓抑制小鼠造血调控的影响

Effect of Zuogui pill on hematopoietic regulation of myelosuppressed mice

目的观察左归丸对骨髓抑制小鼠外周血、造血干细胞增殖能力、骨髓细胞周期和凋亡的影响,并探讨其可能的造血调控机制。方法将48只大鼠随机分为6组,即正常对照组、模型组、阳性对照组和左归丸低、中、高剂量组,各8只,除正常对照组外,其余各组采用60Coγ射线和环磷酰胺联合制作骨髓抑制大鼠模型,于造模结束后第2d开始给药,模型组、正常对照组予0.9%氯化钠注射液,左归丸低、中、高剂量组予相应浓度左归丸,阳性对照组予重组人粒细胞集落刺激因子注射液。各组均给药7d。用全自动血细胞分析仪、白细胞计数法、造血祖细胞培养和流式细胞术分别检测左归丸对骨髓抑制小鼠外周血、骨髓有核细胞数、体外培养各系造血祖细胞的集落数以及骨髓细胞周期和调亡的影响。结果左归丸中、高剂量均能明显升高外周血红细胞、血红蛋白、骨髓有核细胞(P<0.05,P<0.01),左归丸中剂量对血小板和左归丸高剂量对白细胞有明显升高作用(P<0.05)。左归丸中剂量组和阳性对照组能提高体外培养各系造血祖细胞的集落数(P<0.05,P<0.01),左归丸高剂量组粒单系细胞集落生成单位、红系细胞集落生成单位和爆增型红细胞集落生成单位数提高显著(P<0.05,P<0.01)。左归丸各剂量组均能促进骨髓G0/G1期细胞向S期细胞以及S期细胞向G2/M期细胞的转化,从而导致G2/M期细胞比例和增殖指数(PI)明显升高(P<0.05,P<0.01)。左归丸各剂量组的骨髓细胞凋亡比例与模型组比较均显著下降(P<0.01),与阳性对照组比较差异无统计学意义(P>0.05)。结论左归丸可能通过促进G0期造血干细胞进入细胞周期,进行增殖;加速骨髓细胞修复受损的DNA,通过G1/S和S期监测点;抑制造血细胞的凋亡等机制,从而促进损伤骨髓造血功能恢复。

Objective To examine the effect of Zuogui pill on peripheral blood cells, survival and proliferation of hematopoietie stem cells （HSCs） as well as cell cycle of BMCs in myelosuppressed mice, and to explore its mechanism in hematopoietic regulation. Methods 48 mice were randomly divided into six groups, normal control group, model group, positive control group, low - dose, middle - dose and high - dose Zuogui pill group. Except normal control group, myelosuppressed mice were established by 60Co γ ray and eyclophosphamide in other five groups. Mice in normal control group received 0.9% sodium chloride, mice in low - dose, middle - dose and high - dose Zuogui pill group received low - dose, middle - dose and high - dose Zuogui pill, respectively. Mice in positive control group received recombinant human granulocyte colony - stimulating factor injection. The course was 7 day in each group. The effect of Zuogui pill on peripheral blood cells, BMCs , survival and proliferation of HSCs and cell cycle of BMCs in myelosuppressed mice was detected by automatic blood cell analysator, white blood count, hemopoi- eric progenitor cell culture and flow cytometry （FCM） respectively. Results The results showed that medium and high- dose Zuogui pill obviously elevated red blood cells,hemoglobin and BMCs;medium- dose Zuogui pill had ob- viously effect on blood platelet. While high -dose Zuogui pill significantly elevated white blood cells; Medium and high - dose Zuogui pill could obviously pro- mote the Colony forming unit - erythroid （ CFU - E）, Colony forming unit- granulocytemacrophage （ CFU - GM ）, Colony forming unit - megakaryo- cyte （CFU- Meg）. Three- dose Zuogui pill all effectively promoted the transition of BMCs from GO / G1 to S phase and S to G2 / M phase, and obviously elevated the proliferation index;Additionally Zuogui pill can reduce the ratio of apoptosis of BMCs as significantly as positive control group. Conclusion These data suggest that Zuogui pill may promote the recovery of hematopoietic function of the myelosuppressed mice by activating the en- try of HSCs from GO phase into cell cycle, promoting the repairment of the damaged DNA, accelerating GO/G - S phase and S - G2/M phase transition, and inhibiting apoptosis of BMCs.