珍宝丸预处理对新生大鼠缺氧性脑损伤Fas和Fas L影响的实验研究

Experimental study of treasure pill pretreatment on Fas and Fas L of neonatal rats hypoxic brain injury

目的:探讨珍宝丸预处理对新生大鼠缺氧性脑损伤Fas和Fas L蛋白表达的影响。方法:取新生7 d Wistar大鼠200只,雌雄不限,随机分成5组(n=40):正常对照组、模型缺氧1 h组(Ⅰm-12 h、Ⅰm-24 h、Ⅰm-48 h、Ⅰm-72 h)、模型缺氧4h组(Ⅳm-12 h、Ⅳm-24 h、Ⅳm-48 h、Ⅳm-72 h)、珍宝丸缺氧1 h治疗组(ⅠT-14 h、ⅠT-24 h、ⅠT-48 h、ⅠT-72 h)、珍宝丸缺氧4 h治疗组(ⅣT-14 h、ⅣT-24 h、ⅣT-48 h、ⅣT-72 h)。将新生Wistar大鼠置于8%O2+92%N2(V/V)的缺氧箱内建立新生鼠缺氧模型。在造模前每日一次连续7 d灌胃给予生理盐水和珍宝丸,缺氧期间也同样剂量给药,分别于缺氧后12、24、48、72 h处死动物取血及脑组织待测。采用ELISA法检测大鼠血清及脑组织悬液中Fas和Fas L的蛋白含量;采用免疫组织化学方法检测各组大鼠脑组织中Fas和Fas L蛋白的阳性表达。结果:与正常对照组比较,其他四组Fas和Fas L蛋白含量、蛋白的阳性表达均在缺氧后12 h时间点明显增高,且在24 h时间点达高峰,有统计学意义(P<0.01),模型组在各时间点上明显高于治疗组;模型组在48、72 h时间点Fas和Fas L表达低于12 h时间点,但仍明显高于正常对照组(P<0.01);珍宝丸治疗组在48、72 h时间点Fas和Fas L表达低于12 h时间点,但仍高于正常对照组(P<0.05)。结论:珍宝丸可通过下调缺氧性脑损伤大鼠Fas和Fas L的蛋白含量、蛋白的阳性表达强度,从而对缺氧性脑损伤具有一定的保护作用。

To investigate the effect of treasure pill pretreatment on hypoxic brain injury Fas and Fas L protein expression.Methods：Newborn 7d Wistar rats 200,male or female,were randomly divided into 5 groups（n=40）：normal control group, model group 1h hypoxia（Ⅰm-12h,Ⅰm-24h,Ⅰm-48h,Ⅰm-72h）,model group 4h hypoxia（Ⅳm-12h,Ⅳm-24h,Ⅳm-48h,Ⅳm-72h）, hypoxic 1h treasure pill treatment group（ⅠT-14h,ⅠT-24h,ⅠT-48h,ⅠT-72h）,hypoxic 4h treasure pill treatment group（ⅣT-14h,ⅣT-24 h ,ⅣT-48 h ,ⅣT-72 h ）.The newborn Wistar rats were placed in hypoxia chamber of 8%O2＋92%N2 （ V/V） for establishment the model of neonatal rats with hypoxia.Before modeling continuous 7 d once-daily oral administration of saline and treasure pill , during hypoxia also dose ,after hypoxia 12 h,24 h,48 h,72 h animals were sacrificed blood and brain tissues tested.By ELISA to detect the protein levels of Fas and Fas L in serum and brain tissue;by immunohistochemistry method to detect the expression of Fas and Fas L in rat brain tissue.Results：Compared with normal control group ,other four groups the protein content and protein expression of Fas and Fas L are in the 12h time points after hypoxic significantly increased ,and 24 h time point that pealed at statistically significant （ P〈0.01 ） ,model group was significantly higher than the treatment group at each time point ;model groups in 48 h,72 h time point the protein expression and protein content of Fas and Fas L was less than 12 h time point but still significantly higher than the control group （ P〈0.01 ）;treasure pill groups in 48 h,72 h time point the protein expression and protein content of Fas and Fas L was less than 12 h time point ,and slightly higher than the normal control group （ P〈0.05 ）.Conclusion： Treasures pill can cut hypoxic brain damage in rats,the positive expression of Fas and Fas L protein content and protein ,and thus the hypoxic brain injury has a protective effect.