CN106074562A - 桦木酸作为机体镉清除剂的应用 - Google Patents
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Abstract
本发明涉及桦木酸的新用途,尤其涉及桦木酸在制备清除机体内镉的药物、保健品及食品中的应用。本发明通过体外及体内实验证实桦木酸对正常细胞及小鼠没有明显的毒副作用,能减少小鼠肝、肾和睾丸中的镉残留量,能促进尿液中镉的排出;能减轻镉造成的小鼠肝肾和睾丸组织损伤。桦木酸具备治疗机体镉积累的作用,安全有效,优于传统的治疗药物,从而扩宽了其应用范围,也为治疗重金属镉中毒提供了新途径。
Description
技术领域
本发明涉及桦木酸的新用途,具体涉及桦木酸在制备清除机体内镉的药物中的应用。
背景技术
镉是银白色有光泽的金属,镉不是人体的必需元素。人体内的镉是出生后从外界环境中吸取的﹐普通人镉接触主要有四个途径:空气,水,食物和烟草。80%的食物镉来自于谷类和蔬菜,这些食物源于富集镉的土壤;另外由于烟草能够选择性地从土壤中吸收镉,吸烟者血液中血镉的浓度明显大于不吸烟者。镉进入体内以后,由于其10-30年的生物半衰期以及极低的体外排出率,因此对人体产生很大的毒性。体内的镉约50%富集在肝和肾里,对肝和肾的损害极大,表现为尿中含大量低分子量蛋白,由于肾小管功能受损,造成钙、磷和维生素D代谢障碍,进而造成骨质软化和疏松,严重者极易发生病理性骨折;其余分布于肺﹑胰﹑甲状腺﹑睾丸﹑毛发等处。相关的流行病学研究,提示慢性镉中毒患者可能出现神经系统、免疫系统、生殖系统损害等。镉污染的现实危害,在于目前还没有特效的解毒药物可供使用。
目前报道过的作用于镉中毒的药物主要包括:1)传统的驱镉药物:氨基羧酸类化合物EDTA,巯基类化合物和DDTC类化合物,此类药物药物选择性不高,长时间使用有较强的脑、肾毒副作用。2)维生素B和维生素C和E,通过抗氧化起一定的保护作用,不能促镉排出。3)锌、铁、钙和硒等营养素防治,与特定蛋白结合来拮抗镉的毒性,长期摄入钙会有精神副作用,锌过量会导致免疫能力伤害,铁作用于转运蛋白,受蛋白量的限制;此方式拮抗镉的效率有限,并且并不能减少组织内镉的残留量。
桦木酸(betulinic acid,BA)是一种萃取自白桦树的五环三萜类化合物,生物效应特别广泛,目前研究的热点主要集中在桦木酸的抗肿瘤、抗艾滋、抗病毒、抗炎症、抗疟原虫,特别是在抗肿瘤方向。实验已经证实桦木酸对肿瘤细胞有很强的杀伤作用而对正常细胞几乎没有副作用。
目前,尚未见关于桦木酸用于治疗重金属镉中毒的报道。
发明内容
为了解决现有技术中存在的问题,本发明的第一个目的是提供桦木酸在制备清除机体内镉的药物中的应用。
本发明的第二个目的是提供桦木酸在制备清除机体内镉的保健品中的应用。
本发明的第三个目的是提供桦木酸在制备清除机体内镉的食品中的应用。
以上目的通过如下实验加以验证:
1)体外实验:用桦木酸和氯化镉联合处理三种正常的肝、肾和睾丸细胞,通过CCK-8细胞增殖实验和流式细胞仪检测细胞凋亡情况,确定最佳给药时间和给药浓度;同时观察到桦木酸能在一定程度上拮抗氯化镉引起的细胞凋亡。
2)体内实验:
(1)昆明小鼠分成四组,一组空白对照组,其他三组分别为3mg/kg,10mg/kg和30mg/kg三个剂量桦木酸组,灌胃。观测体重变化,10天后处死小鼠,分析数据,检测HE染色,确定桦木酸在ABC三个剂量对小鼠的毒副作用。
(2)昆明小鼠分成五组,空白对照组,氯化镉模型组和三组不同剂量桦木酸预处理组,氯化镉采取1 mg/kg浓度腹腔注射。每天提前1小时进行灌胃处理再腹腔注射,观测体重变化,连续10天,然后饥饿24小时,处死小鼠。HE染色,石墨原子吸收光谱,分析数据,确定桦木酸预处理对小鼠镉中毒的保护效果。
结论:桦木酸在3mg/kg,10mg/kg和30mg/kg三个剂量对小鼠没有明显的组织毒副作用,仅轻微减轻小鼠体重;桦木酸能减少小鼠肝、肾和睾丸中的镉残留量,能促进尿液中镉的排出;能减轻镉造成的小鼠肝肾和睾丸组织损伤。
本发明的积极进步效果在于:首次发现了桦木酸对机体镉积累所致的组织损伤的保护作用;在治疗机体镉积累时,无组织毒副作用,能减少组织的镉残留,减少镉所致的组织损伤,安全有效,优于传统的治疗药物,从而扩宽了其应用范围,也为治疗重金属镉中毒提供了新途径。
附图说明
图1不同剂量氯化镉对TM3、293T和LO-2细胞增殖的影响图;
图2不同剂量桦木酸对TM3、293T和LO-2细胞增殖的影响图;
图3桦木酸、氯化镉及桦木酸与氯化镉同时作用对细胞凋亡的影响图;
图4不同剂量桦木酸(BA)对小鼠体重和脏器系数的影响图;
A为不同剂量的桦木酸对小鼠体重的影响;B为不同剂量的桦木酸对小鼠睾丸脏器系数的影响;C为不同剂量的桦木酸对小鼠肝脏脏器系数的影响;D为不同剂量的桦木酸对小鼠肾脏脏器系数的影响;
图5不同剂量桦木酸对小鼠肝肾生化指标的影响图;
A为不同剂量的桦木酸对小鼠谷草转氨酶的影响;B为不同剂量的桦木酸对小鼠谷丙转氨酶的影响;C为不同剂量的桦木酸对小鼠尿素氮的影响;
图6不同剂量桦木酸对小鼠肝组织病理变化的影响;
A为空白对照;B为3 mg/kg BA组;C为10 mg/kg BA组;D为30 mg/kg BA组;
图7不同剂量桦木酸对小鼠肾组织病理变化的影响;
A为空白对照;B为3 mg/kg BA组;C为10 mg/kg BA组;D为30 mg/kg BA组;
图8不同剂量桦木酸对小鼠睾丸组织病理变化的影响;
A为空白对照;B为3 mg/kg BA组;C为10 mg/kg BA组;D为30 mg/kg BA组;
图9不同剂量桦木酸预处理对氯化镉中毒小鼠体重与脏器系数的影响图;
A不同剂量的桦木酸预处理对镉中毒小鼠的体重的影响
B不同剂量的桦木酸预处理对镉中毒小鼠肝脏脏器系数的影响
C不同剂量的桦木酸预处理对镉中毒小鼠肾脏脏器系数的影响
D不同剂量的桦木酸预处理后小鼠睾丸的直接形态大小
E不同剂量的桦木酸预处理对镉中毒小鼠睾丸脏器系数的影响
图10不同剂量桦木酸预处理对镉中毒小鼠肝肾睾丸组织中镉残留量与尿液中镉含量的影响图;
A小鼠肝脏中镉残留量;C小鼠睾丸中镉残留量;B小鼠肾脏中镉残留量;D小鼠尿液中镉含量;
图11不同剂量桦木酸预处理对镉中毒小鼠肝组织病理变化的影响图;
图12不同剂量桦木酸预处理对镉中毒小鼠肾组织病理变化的影响图;
图13不同剂量桦木酸预处理对镉中毒小鼠睾丸组织病理变化的影响图;
图14不同剂量桦木酸预处理对镉中毒小鼠肝肾生化指标的影响图。
具体实施方式
通过以下详细说明结合附图可以进一步理解本发明的特点和优点。所提供的实施例仅是对本发明方法的说明,而不以任何方式限制本发明揭示的其余内容。
本发明中,所述的桦木酸为植物白桦树中提取的三萜类化合物,市售可得。本实验中,昆明小鼠皆来自湖北省疾病控制中心的洁净鼠。293T:正常肾上皮细胞、LO-2:正常肝细胞、TM3:正常睾丸间质细胞,均来自武汉大学中国典型培养物保藏中心。桦木酸溶解于玉米油中灌胃处理,氯化镉进行腹腔注射。桦木酸分为低中高三个剂量,氯化镉采取低剂量注射。除特殊说明外,本发明所用试剂和原料均市售可得。
【实施例1】桦木酸对氯化镉诱导正常细胞凋亡的影响
通过CCK-8细胞增殖实验从时间梯度和浓度梯度检测氯化镉对三种正常细胞TM3、293T和LO-2细胞的活性抑制,确定急性低剂量中毒的最佳给药剂量和作用时间,然后用这个时间梯度来选择最佳给药桦木酸浓度,再用桦木酸和氯化镉联合作用于细胞,流式检测。
1、不同剂量氯化镉对TM3、293T和LO-2细胞增殖的影响
三种正常细胞TM3、293T和LO-2细胞与不同浓度氯化镉溶液分别孵育12h、24h和48h后CCK-8检测细胞活性,所得结果如图1。从图中可以看到,24小时,氯化镉在10 μm/l对三种正常细胞产生了明显的活性抑制,48小时时这种抑制更加明显,由于本发明偏重于急性中毒,因此,镉的急性中毒模型采用氯化镉的浓度为10 μm/l,作用24小时。
2、不同剂量桦木酸对TM3、293T和LO-2细胞增殖的影响
由图2可见,不对三种正常细胞产生抑制作用的最大桦木酸浓度,为20mg/ml。因此,在本发明的实施例中采用这个浓度处理镉中毒的细胞或小鼠模型。
3、桦木酸、氯化镉及桦木酸与氯化镉同时作用对细胞凋亡的影响
细胞计数后铺六孔板,分为空白对照组、桦木酸对照组(20mg/ml)、氯化镉对照组(10μm/l),及给药20mg/ml的桦木酸1h后,给药10μm/l的氯化镉组,一起孵育24小时,流式细胞仪检测细胞凋亡情况。从图3中可以看到,桦木酸几乎不促进细胞凋亡,氯化镉能明显导致细胞凋亡,桦木酸和氯化镉联合孵育细胞,细胞凋亡情况有明显的减轻,因此,桦木酸能拮抗氯化镉所致的细胞凋亡。
【实施例2】桦木酸毒副作用的检测
四组昆明小鼠共24只每组6只,一组空白对照组,其他三组分别为3mg/kg,10mg/kg和30mg/kg三个剂量桦木酸组,灌胃。每天记录小鼠的体重,10天后处死小鼠,称取脏器重量,获得脏器系数,脏器系数=(器官重量/小鼠体重)*1000;分析生化数据,检测HE染色,确定桦木酸在三个剂量对小鼠的毒副作用。
1、不同剂量桦木酸(BA)对小鼠体重和脏器系数的影响
从图4中可以看到,桦木酸在30mg/kg剂量内仅轻微降低小鼠体重,但是对小鼠肝、肾和睾丸的脏器系数并没有影响(脏器系数增大,脏器肿大;脏器系数下降,脏器发育不良),与空白对照组相比没有明显变化。
2、不同剂量桦木酸对小鼠肝肾生化指标的影响
从图5A和B可以看出,在给药不同剂量的桦木酸后,小鼠肝脏的生化指标谷草转氨酶和谷丙转氨酶与对照组相比几乎没有明显的变化;在C图中,可以看到给药组的尿素氮与空白对照组相比几乎没有明显的变化。
3、不同剂量桦木酸对小鼠肝、肾及睾丸组织病理变化的影响
从图6中可以看到,三个剂量给药组B、C、D与空白对照组A相比,细胞排列整齐规则,细胞核明显(箭头所示),细胞碎片没有增加,无病变出现;肝中央静脉腔无萎缩混乱。
从图7中可以看出,三个剂量给药组B、C、D和空白对照组A相比,肾小囊腔没有萎缩,肾小球无萎缩出现,远端小管处细胞没有排列混乱,周围细胞排列规则有序。无明显的病变情况出现。
从图8中可以看出,三个剂量给药组B、C、D和空白对照组A相比,生精细胞排列规则有序,各级生精细胞皆有,腔中成熟精子数量皆很多。
总之,桦木酸在3mg/kg,10mg/kg和30mg/kg三个剂量对小鼠几乎没有的肝肾和睾丸组织毒副作用。
【实施例3】桦木酸预处理对镉中毒小鼠的保护效果
五组昆明小鼠30只每组6只,空白对照组,氯化镉模型组和三组不同剂量桦木酸预处理组,灌胃。氯化镉采取1 mg/kg浓度腹腔注射。每天提前1小时进行灌胃处理再腹腔注射,连续10天,然后饥饿24小时,处死小鼠,HE染色,石墨原子吸收光谱检测镉的残留量,分析数据。确定桦木酸预处理对镉中毒小鼠的保护效果。
1、不同剂量桦木酸预处理对氯化镉中毒小鼠体重与脏器系数的影响
从图9A可以看到,十天处理后,氯化镉能明显降低小鼠的体重,桦木酸能拮抗氯化镉引起的这种下降,并且呈剂量正依赖。9B和9C中可以看到,氯化镉能明显增加小鼠肝肾的脏器系数,桦木酸能明显降低这种增加,呈剂量依赖正相关。在9D图中,十天处理后氯化镉能明显减小小鼠睾丸的大小,氯化镉能降低小鼠睾丸的脏器系数如图9E所示,桦木酸能拮抗这种降低。氯化镉引起睾丸脏器系数下降,可能是由于实验采用的是未完全性成熟的小鼠,在实验过程中,性腺发育被抑制。
2、不同剂量桦木酸预处理对镉中毒小鼠肝肾睾丸组织中镉残留量与尿液中镉含量的影响
肝、肾和睾丸组织在小鼠处死当天送去做石墨原子吸收光谱。由于镉在生物体内极难排出,因此取第十天的小鼠尿液检测尿液中的镉含量。从图10A、B、C中可以看到桦木酸有效降低小鼠肝、肾和睾丸组织中镉的含量,并且呈剂量依赖。从图10D中可以看到,桦木酸能增加尿液中镉的排出,呈剂量依赖。
3、不同剂量桦木酸预处理对镉中毒小鼠肝组织病理变化的影响
从图11中可以看到,氯化镉十天处理后,肝的中央静脉和门管区出现了明显的病变;但是随着桦木酸剂量的增加,肝的门管区和中央静脉的病变情况逐渐降低,在30mg/kg时恢复到与空白对照组接近的水平。
4、不同剂量桦木酸预处理对镉中毒小鼠肾组织病理变化的影响
从图12中可以看到,氯化镉十天处理后,肾小球和肾小管出现了明显的病变,细胞排列胡乱无序,大量氧化空泡变性。但是随着桦木酸剂量的增加,肾的肾小管和肾小球的病变情况逐渐降低,氧化空泡减少,细胞逐渐规则,在30mg/kg时恢复到与空白对照组接近的水平。
5、不同剂量桦木酸预处理对镉中毒小鼠睾丸组织病理变化的影响
从图13中可以看到,氯化镉十天处理后,随BA剂量增加,睾丸生殖上皮细胞的病理状态逐渐减轻,表现出明显的量效关系。
6、不同剂量桦木酸预处理对镉中毒小鼠肝肾生化指标的影响
试剂盒检测小鼠的肝肾生化指标结果如图14A、B、C所示,在图中,氯化镉能引起谷草转氨酶,谷丙转氨酶和尿素氮的明显上升,但是桦木酸预处理后,这种变化会降低,并且呈剂量依赖正相关。
结论:桦木酸预处理能拮抗氯化镉所致的小鼠体重的下降,能减小氯化镉诱导产生的肝、肾和睾丸组织损伤,起一定的保护作用,保护效果与剂量呈依赖正相关。能有效降低小鼠肝、肾和睾丸组织中镉的残留量,能增加尿液中镉的排出量,残留量与排出量皆与剂量呈依赖正相关。
Claims (3)
1.桦木酸在制备清除机体内镉的药物中的应用。
2.桦木酸在制备清除机体内镉的保健品中的应用。
3.桦木酸在制备清除机体内镉的食品中的应用。
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-
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Non-Patent Citations (2)
Title |
---|
AGNIESZKA SZUSTER-CIESIELSKA ET AL.: ""Protective effects of betulin and betulinic acid against ethanol-induced cytotoxicity in HepG2 Cells"", 《PHARMACOLOGICAL REPORTS》 * |
SEON-HEE ET AL.: ""Protection of betulin against cadmium-induced apoptosis in hepatoma cells"", 《TOXICOLOGY》 * |
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