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Experimental Research on the Effects of YinLing Ⅰ on Expelling Lead and Improving Non-special Immune Function in Lead-poisioned Mice

Author: ShengKai
Tutor: ZhangGuoCheng；XuDongLiang
School: Fourth Military Medical University
Course: Pediatrics
Keywords: YinLing I Lead poisoning therapy
CLC: R285.5
Type: Master's thesis
Year: 2005
Downloads: 110
Quote: 1
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Abstract


Lead poisoning is the most common heavy metal intoxication and can lead to multi-organ injury if entering body. Children has a high incidence of lead poisoning because of their habits and characteristics. Presently the chelated preparations,such as EDTANa2Ca and NaDMS,is abroadly adopted to treat lead poisoning, but they have many side-effects though they have good curative effect. Recently,Chinese traditional medicine is developed to treat lead poisoning.YinLing I is a primary filtrated herb preparation ,made from gingkgo,tuckahoe,etc.Its efficacy is to invigorate Qi, to strengthen spleen, to nourish kidney , to soothe the nerves and so on. This study is to investigate the effects of YinLing I on expelling lead and improving non-special immune function in lead-poisioned mice and the effects of YinLing I on the cell viability and oxidative injury of lead-exposed cell.[Methods]1. The 48 KunMing mice were randomly divided into four groups: the control group and three groups with lead posioning （Pb control group, YinLing I group, EDTA group）. The mice from three groups with lead poisoning were exposed to 0.2% lead acetate in drinking water ad libitum for 10 weeks. After 10-week lead exposure the control group and the Pb control group were given water, and the other two groups were respectively administered YinLing I and EDTANa2Ca for 8 weeks. The concentrations of lead in the blood, brain, liver, kidney and bone and the concentrations of zinc, calcium, copper in the blood were quantified; the ability of learning and memory of mice was measured monthly by Y-water maze test; ultrastructure of specimen from each group was observed and analyzed with light microscope and transmission electron microscope.2. The 32 KunMing mice were grouped and exposed to lead as the above experiment. After 3-week administration of YinLing I and EDTANa2Ca,the number of white blood cell, the rate of carbon expurgation, the coefficient of immune organ of mice were detected to evaluate the non-special immune function.3. After lead exposure Vero cell was treated with YinLing I of different concentrations. The cell viablitity was measured by MTT method and the SOD and MDA activity in cell suspensions were measured.[Results] 1. Effect of YinLing I on the concentration of lead in the blood and organs inlead-posioned miceAfter 10-week lead exposure the blood lead concentrations of three groups with lead poisoning （456.67 ±49.19ug/L、 450.67± 63.57ug/L、 453.92 ± 58.35ug/L） was significantly higher than that of the control group（28.50± 5.05ug/L） （P<0.01） .The lead concentrations in the blood（162.50± 21.72 ug/L）,brain（23.42 ± 6.73ug/L）,liver（43.17 ± 20.08ug/L）,kidney（24.83 ± 5.94ug/L）and bone（345.25±47.42ug/L） were significantly suppressed by the administration of YinLing I （P<0.05）, and no evident difference was found in the lead concentration of the brain （20.42±7.43ug/L）、 and liver（24.92± 8.63ug/L） between the YinLing I group and EDTA group（P>0.05）.2. Effect on the concentrations of zinc, calcium, copper in the bloodThe concentrations of zinc（101.93 ± 17.87umol/L）,calcium（1.50 ± 0.21mmol/L）,copper（21.92 ± 5.65umol/L） in the blood did not show any change after treatment with YinLing I .Compared with YinLing I ,the concentrations of zinc（77.3 + 7.05umol/L）, and copper（12.79 + 5.47umol/L） in the blood were significantly suppressed by EDTA（P<0.05）.3. Effect of YinLing I on ability of learning memory in lead-poisioned miceIn Y-water maze test, the mice with lead poisoning had much less correct reaction times（1. 8 ± 1. 62） 、 and longer time of arrival（18. 20 + 2. 30s） compared with the control group（P<0.0l）.The 4-week and 8-week administration of YinLing I increased the correct reaction times and shortened the arrival time, and after 8-week administration YinLing I improved significantly the above index compared with EDTA（P<0.05）.4. Effects of YinLing I on ultrastructure of organs4.1 light microscopeThe HE staining of the specimen showed:（T）The control group:the kidney, liver,brain showed the normal ultrastructure.（2）Pb control group:in the kidney blood vessel was congested, glomeruli atrophied, the cell of tubule swelled and tubule dilated. The structure of the liver lobule was disordered, the liver cell swelled and inflammational cell soaked in the liver cell. The nerve cell of the brain cortex shrinked or swelled and the nerve cells of hippocampi crimpled and were unorderly arranged. ③YinLing I group: the pathological manifestions of the kidney, liver and brain were ameliorated to the different extent with the administration of YinLing I .④ EDTA group: the pathological manifestions of the liver and brain were improved, but the kidney showed the dilation of blood vein and focal fibrosis.4.2 transmission electron microscope① the control group: the epithelium of the renal tubule, liver cell, the cell of brain cortex and hippocampi showed the normal mitochondrial and endoplasmic reticulum and had abundant organelles.②Pb control group: there was significant anomaly in Pb control group,with the organelles incytoplasm of pyramidal cell decreasing,mitochondrial swelling, endoplasmic reticulum enlarging , but lipofuscin increasing in number, vacuaoltion being found in myeliuated nerve fiber axon,and astrocyte in vessels edemaing. ③YinLing I group:the swelling mitochondrial and dilated endoplasmic reticulum become little,organelles increased, lipofuscin decreased,the swellingastrocytes were reduced.④EDTA group:there were also the improvedultrastructure of the liver and brain except the kidney.5. Effects of YinLing I on the non-special immune function in lead-poisioned miceThe number of white blood cell（10. 11 ± 1. 40× 109/L）, the rate of carbon expurgation（K=0. 82 ± 0. 43; α =4. 08 ± 0. 86）, the coefficient of immune organ（SI=2. 64 ± 0. 26 ; TI=1. 94 ± 0. 46） decreased after lead exposure. YinLing I ameliorated the above index,which is （12. 34 ± 0.90× 109/L）、 （K=2.25 ± 0.82; α=5.43±1.07）、 （SI=3. 64 ± 0. 56; TI=2. 75±0. 48）; and had a better effect than EDTA.on the coefficient of immune organ, which is11. 71 ± 1.34 × 109/L , （K=l. 32 ± 1.20 ; α =4.15 ±1. 20）and（SI=3. 22 + 0. 28; TI=2. 41±0. 38）.6.Effects of YinLing I on the cell viability and oxidative injury of lead-exposed cellYinLing I can increase the viability of lead-exposed cell during a certain extent; in the most non-toxicity concentration YinLing I can elevate the SOD content in the Vero cell suspension, reduce the MDA concentration and resisting lead intoxication in vitro. [Conclusions]This study utilized in vivo and in vitro experiments to investigate the effects of YinLing I on lead-poisioned mice. The study showed: 1. YinLing I can reduce the concentrations of lead in the blood, brain, liver, kidney and bone.

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