苞叶雪莲水提物对小鼠免疫系统辐射防护作用的研究
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摘要
研究目的
观察苞叶雪莲水提物对辐射损伤小鼠免疫功能的调节作用。
研究方法
免疫实验：通过游泳实验及胸腺、脾脏重量测定并指数计算,鸡红细胞作免疫原的溶血素测定实验、PHA刺激淋巴细胞转化实验和腹腔巨噬细胞功能测定实验来反映受照射小鼠的免疫功能及全身状态。实验小鼠共分为五组(每组8只)：正常对照(Control)、照射对照(IR)、药物低(IR+LD)、中(IR+MD)、高剂量组(IR+HD)。用药组予不同剂量苞叶雪莲水提物灌胃,对照组予等量生理盐水灌胃。用4MV-x ray照射造模,预防性给药(照射前)一周,治疗性给药(照射后)一周,共14天。后按既定方法对各组小鼠进行指标检测。微核实验：照射后24h处死小鼠,观察骨髓微核率。所有实验数据用spss11.5统计软件进行统计分析。
研究结果
1.照射对小鼠的免疫功能及全身状态均有明显的影响。小鼠体重增长减慢、游泳时间减短、各项免疫指标均有下降。在溶血素测定实验中IR组小鼠意外死亡两只,其余小鼠均存活。
2.在游泳实验中,Control组游泳时间明显长于其他各组,与IR组比较具有统计学差异,P<0.05,各个用药组游泳时间均长于IR组,但不具有统计学差异。
3.免疫器官称重,脾脏重量及脾脏指数检测结果显示,Control组高于其他各组,用药组高于IR组,但差异无统计学意义。胸腺重量,Control组高于其余各组,用药组高于IR组,用药组中以高剂量组胸腺重量最高。统计学结果显示,Control组与IR组、IR+LD组及IR+MD组比较,差异具有统计学意义,P<0.05,而Control组与IR+HD组比较则无统计学差异。胸腺指数,Control组高于其他各组,用药组高于IR组,IR+HD组最为明显。统计学结果显示,Control组与IR组比较,差异具有统计学意义,P<0.05；IR+HD组与IR组比较存在统计学差异,P<0.05。
4.溶血素测定(OD值),Control组明显高于其他组。统计分析结果显示,Control组与其他各组比较差异均具有统计学意义,P<0.05,其余各组之间无统计学差异。
5.PHA刺激淋巴细胞转化实验及腹腔巨噬细胞功能测定实验结果显示,受照射小鼠血液中淋巴细胞及腹腔巨噬细胞数量明显减少,未形成有效的统计学数值。Control组淋巴细胞转化率为28.7%。
6.微核实验,照射及用环磷酰胺都是小鼠骨髓微核产生明显增多,各个用药组微核产生率较模型组明显降低,中／高剂量组与模型组比较有统计学差异,P<0.05/P<0.01。
结论
1.苞叶雪莲水提物灌胃用药对小鼠免疫系统可能有一定的放射防护作用,对受照射小鼠全身状态有一定的调节作用,表现在：1)各个用药组小鼠游泳时间均长于照射对照组小鼠。2)用药组小鼠脾脏、胸腺重量及其指数,均高于照射对照组,尤其高剂量用药组。
2.微核实验结果表明,苞叶雪莲水提物对小鼠DNA有明显的辐射防护作用,且有一定的剂量依赖性。在药物制备、成分分析、用药方式及剂量、用药时间方面值得进一步探讨。
Objective
To investigate the radioprotective effects of Aqueous Extract of Sauseurea Obyallata (ESO) on immune system of mice from radiation injury.
Methods
Immune function tests:Four subexperiments including swimming test with weighting spleen, thymus and computing their indexs, hemolysin test, PHA-induced lymphocyte transformation test and assay for phagocytosis of peritoneal macrophage were used in the study. Mice with each subexperiment divided into 5 groups:Control group, Irradiation group (IR), Irradiation+high dose group (IR+HD), Irradiation+middle dose group(IR+MD), Irradiation+low dose group (IR+LD). ESO treated animals received intragastric administration of drugs for 14 days, including 7days before irradiation and 7days after irradiation. Other mice received the same amount of Normal Saline. All mice were irradiated with 6 Gy of x-ray except
Control group.
Micronucleus test:24h after irradiation, mice were sacrificed and measured micronucleus.
Data were analyzed by SPSS 11.5.
Results
1. Irradiation have obvious influence on immune function and systemic state of mice.Body weights gowth slowed down, shorten swimming time, each immune index decreased significantly. In hemolysin test, two mice were dead in accident in (?)R group. The remaining mice survived.
2. Swimming test showed that Control group had longer swimming time than other groups. And there was a significant difference between Control group and IR group statistically (P<0.05). ESO treated groups had longer swimming time than IR group, but there was no significant difference between them.
3. Weight of immune organs showed increased splenic weight and splenic index of Control group compared with other groups, but there were no significant difference between them. The thymic weight of Control group was heavier than other groups, and that of ESO treated groups were heavier than IR group. IR+HD group have the heaviest thymus among ESO treated groups. There was a significant difference between Control group and IR group, IR+LD group, IR+MD group, respectively. And no significant difference between Control group and IR+HD group was observed. The thymus index of Control group was higher than other groups. There was a significant difference between Control group and IR group (p<0.05). Thymus indexes of ESO treated groups were higher than IR group, especially HD group had a significantly increased thymus index compared with IR group (P<0.05).
4. Hemolysin test revealed that Control group had a higher OD value than other groups, and there were significant difference between them(p<0.05). There was no obvious difference between ESO treated groups and IR group.
5. PHA-induced lymphocyte transformation test and assay for phagocytosis of peritoneal macrophage revealed that there were not enough Lymphocytes and peritoneal macrophages in mice which received radiation for statistical analysis. The rate of lymphocyte transformation of Control group was 28.7%.
6. Micronucleus test:radiation and CTX caused a significant increase of micronucleus. ESO treated groups have lower Frequency of Micronucleus compared with IR group, especially IR+HD group/IR+MD group (p<0.01/ p<0.05).
Conclusions
1. Aqueous Extract of Sauseurea Obyallata may have radioprotective effects on immune system of mice by intragastric administration, and may have some effects in regulating the body state. The related performances are listed as follow:1) ESO treated groups have longer swimming time IR group.2) ESO treated groups have heavier spleen and thymus weight and higher index than IR group, especially HD group.
2. The result of micronucleus test reflects that ESO has significant radio protective effects on DNA of mice, and in dose-dependent manner. Further research should be focused on compound preparing, chemical composition analysis, using dose, mode and treatment duration of the compound.
观察苞叶雪莲水提物对辐射损伤小鼠免疫功能的调节作用。
研究方法
免疫实验：通过游泳实验及胸腺、脾脏重量测定并指数计算,鸡红细胞作免疫原的溶血素测定实验、PHA刺激淋巴细胞转化实验和腹腔巨噬细胞功能测定实验来反映受照射小鼠的免疫功能及全身状态。实验小鼠共分为五组(每组8只)：正常对照(Control)、照射对照(IR)、药物低(IR+LD)、中(IR+MD)、高剂量组(IR+HD)。用药组予不同剂量苞叶雪莲水提物灌胃,对照组予等量生理盐水灌胃。用4MV-x ray照射造模,预防性给药(照射前)一周,治疗性给药(照射后)一周,共14天。后按既定方法对各组小鼠进行指标检测。微核实验：照射后24h处死小鼠,观察骨髓微核率。所有实验数据用spss11.5统计软件进行统计分析。
研究结果
1.照射对小鼠的免疫功能及全身状态均有明显的影响。小鼠体重增长减慢、游泳时间减短、各项免疫指标均有下降。在溶血素测定实验中IR组小鼠意外死亡两只,其余小鼠均存活。
2.在游泳实验中,Control组游泳时间明显长于其他各组,与IR组比较具有统计学差异,P<0.05,各个用药组游泳时间均长于IR组,但不具有统计学差异。
3.免疫器官称重,脾脏重量及脾脏指数检测结果显示,Control组高于其他各组,用药组高于IR组,但差异无统计学意义。胸腺重量,Control组高于其余各组,用药组高于IR组,用药组中以高剂量组胸腺重量最高。统计学结果显示,Control组与IR组、IR+LD组及IR+MD组比较,差异具有统计学意义,P<0.05,而Control组与IR+HD组比较则无统计学差异。胸腺指数,Control组高于其他各组,用药组高于IR组,IR+HD组最为明显。统计学结果显示,Control组与IR组比较,差异具有统计学意义,P<0.05；IR+HD组与IR组比较存在统计学差异,P<0.05。
4.溶血素测定(OD值),Control组明显高于其他组。统计分析结果显示,Control组与其他各组比较差异均具有统计学意义,P<0.05,其余各组之间无统计学差异。
5.PHA刺激淋巴细胞转化实验及腹腔巨噬细胞功能测定实验结果显示,受照射小鼠血液中淋巴细胞及腹腔巨噬细胞数量明显减少,未形成有效的统计学数值。Control组淋巴细胞转化率为28.7%。
6.微核实验,照射及用环磷酰胺都是小鼠骨髓微核产生明显增多,各个用药组微核产生率较模型组明显降低,中／高剂量组与模型组比较有统计学差异,P<0.05/P<0.01。
结论
1.苞叶雪莲水提物灌胃用药对小鼠免疫系统可能有一定的放射防护作用,对受照射小鼠全身状态有一定的调节作用,表现在：1)各个用药组小鼠游泳时间均长于照射对照组小鼠。2)用药组小鼠脾脏、胸腺重量及其指数,均高于照射对照组,尤其高剂量用药组。
2.微核实验结果表明,苞叶雪莲水提物对小鼠DNA有明显的辐射防护作用,且有一定的剂量依赖性。在药物制备、成分分析、用药方式及剂量、用药时间方面值得进一步探讨。
Objective
To investigate the radioprotective effects of Aqueous Extract of Sauseurea Obyallata (ESO) on immune system of mice from radiation injury.
Methods
Immune function tests:Four subexperiments including swimming test with weighting spleen, thymus and computing their indexs, hemolysin test, PHA-induced lymphocyte transformation test and assay for phagocytosis of peritoneal macrophage were used in the study. Mice with each subexperiment divided into 5 groups:Control group, Irradiation group (IR), Irradiation+high dose group (IR+HD), Irradiation+middle dose group(IR+MD), Irradiation+low dose group (IR+LD). ESO treated animals received intragastric administration of drugs for 14 days, including 7days before irradiation and 7days after irradiation. Other mice received the same amount of Normal Saline. All mice were irradiated with 6 Gy of x-ray except
Control group.
Micronucleus test:24h after irradiation, mice were sacrificed and measured micronucleus.
Data were analyzed by SPSS 11.5.
Results
1. Irradiation have obvious influence on immune function and systemic state of mice.Body weights gowth slowed down, shorten swimming time, each immune index decreased significantly. In hemolysin test, two mice were dead in accident in (?)R group. The remaining mice survived.
2. Swimming test showed that Control group had longer swimming time than other groups. And there was a significant difference between Control group and IR group statistically (P<0.05). ESO treated groups had longer swimming time than IR group, but there was no significant difference between them.
3. Weight of immune organs showed increased splenic weight and splenic index of Control group compared with other groups, but there were no significant difference between them. The thymic weight of Control group was heavier than other groups, and that of ESO treated groups were heavier than IR group. IR+HD group have the heaviest thymus among ESO treated groups. There was a significant difference between Control group and IR group, IR+LD group, IR+MD group, respectively. And no significant difference between Control group and IR+HD group was observed. The thymus index of Control group was higher than other groups. There was a significant difference between Control group and IR group (p<0.05). Thymus indexes of ESO treated groups were higher than IR group, especially HD group had a significantly increased thymus index compared with IR group (P<0.05).
4. Hemolysin test revealed that Control group had a higher OD value than other groups, and there were significant difference between them(p<0.05). There was no obvious difference between ESO treated groups and IR group.
5. PHA-induced lymphocyte transformation test and assay for phagocytosis of peritoneal macrophage revealed that there were not enough Lymphocytes and peritoneal macrophages in mice which received radiation for statistical analysis. The rate of lymphocyte transformation of Control group was 28.7%.
6. Micronucleus test:radiation and CTX caused a significant increase of micronucleus. ESO treated groups have lower Frequency of Micronucleus compared with IR group, especially IR+HD group/IR+MD group (p<0.01/ p<0.05).
Conclusions
1. Aqueous Extract of Sauseurea Obyallata may have radioprotective effects on immune system of mice by intragastric administration, and may have some effects in regulating the body state. The related performances are listed as follow:1) ESO treated groups have longer swimming time IR group.2) ESO treated groups have heavier spleen and thymus weight and higher index than IR group, especially HD group.
2. The result of micronucleus test reflects that ESO has significant radio protective effects on DNA of mice, and in dose-dependent manner. Further research should be focused on compound preparing, chemical composition analysis, using dose, mode and treatment duration of the compound.
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[1]卢开柏,傅源,常欣欣.雪莲的毒理试验研究[J].新疆中医药,1996,2：药26-27.
[2]陈发菊,杨映根,赵德修,等.我国雪莲植物的种类、生境分布及化学成分研究进展[J].植物学通报,1999,16(5)：561-566.
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[4]金越,吕勇,韩国柱,等.槲皮素及异槲皮素、芦丁抗自由基活性的比较研究[J].中草药,2007,38(3)：408-412.
[5]吴征镒.全国中草药汇编[M].北京：人民卫生出版社,1997,755-761.
[6]贾景明,吴春福.天山雪莲培养物对小鼠免疫功能的影响[J].中华中医药杂志(原中国医药学报),2007,22(4)：238-240.
[7]张振明,葛斌,许爱霞,等.雪莲花水提物的抗氧化活性及其机制[J].第二军医大学学报,2005,26(5)：568-570.
[8]高博,粱中琴,顾振纶.天山雪莲水提物的抗辐射损伤机制[J].江苏医药,2003,29(1)：17.
[9]杨若明,蓝叶芬,蓝翁驰等.两种藏药雪莲花的元素测定[J].中央民族大学学报(自然科学版),2005,14(2)：120-123.
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[14]王如阳,刘晓芳,刘满红,等.雪莲果乙醇提取液清除羟自由基的抗氧化性能研究[J].微量元素与健康研究,2008,25(5)：53-54.
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[22]徐进彦,李文辉,刘建波,等.苞叶雪莲水提物对小鼠辐射防护作用研究[J].中国辐射卫生,2008,18(1)：22-23.
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