“自免肌炎”大鼠经天花粉干预后血清肌酶和HMGB1、TOLL样受体传导的改变

发布时间：2018-04-25 23:08

  本文选题：特发性炎症肌病 + 高迁移率族蛋白B1　； 参考：《辽宁中医药大学》2016年硕士论文

【摘要】：目的:研究单味中药天花粉影响实验性自身免疫性肌炎的大鼠血清肌酶(肌酸激酶、乳酸脱氢酶、天门冬氨酸氨基转移酶)和HMGB1、TLR2、TLR4信号传导改变的情况,进一步了解实验性自身免疫性肌炎大鼠经过天花粉干预后,肌肉炎症损伤下的修复情况以及产生效果的机制。材料与方法:1.大鼠组别分配与免疫把四十只SPF级SD大鼠预先在实验室中喂养一星期之后,仿效完全随机设计的方法,把预先喂养的四十只SD大鼠随机的分成四个组别,分别为:空白组、模型组、激素组、天花粉组。每组分别为十只。由异种动物(家兔)的骨骼肌作为肌匀浆的原料。制备好肌匀浆后将肌匀浆和完全弗氏佐剂各等份,把它们均匀搅拌后乳化。其中空白组中十只不用任何方法干预,剩下的三十只以之前调对好的等比例肌匀浆和完全弗氏佐剂进行免疫注射。注射的位置选取在大鼠背部的左右双侧,实行皮下注射,每次注射量均为0.35毫升,每星期一次,此次注射后于第七天,第十四天,第二十一天,第二十八天,第三十五天对大鼠再进行注射,经6次注射后,完成对大鼠的免疫。2.给药观察与提取样本分析在免疫四个星期后,大鼠造模成功以后进行药物干预。空白组和模型组采用生理盐水来灌胃,而剩下两组均采用每一组特有的处理方式进行灌胃。四组一共灌胃四周,四周后称重大鼠的体重,分组记录数据,待体重测定完后麻醉,从腹主动脉中取血作为血液样本,采集肌肉等相关组织作为肌肉样本,后处死。3.处理样本与分析样本通过Lennon LA的评分细则,评定经过药物干预后大鼠的临床状态,并加以评分;对最后一次灌胃后四组之间的体重进行对比;鉴定血清中肌酸激酶、乳酸脱氢酶、天门冬氨酸氨基转移酶的值;通过酶联免疫吸附法,获得大鼠肌肉里HMGB1、TLR2、TLR4的值,并加以鉴定。结果:1.天花粉可以改善肌酸激酶(CK),乳酸脱氢酶(LDH),天门冬氨酸氨基转移酶(AST)在血清中的含量,起到降低CK、LDH、AST的作用。2.天花粉对HMGB1、TLR4传导产生影响,对TLR2传导未产生影响。天花粉可减少HMGB1、TLR4在肌肉中的含量。结论:天花粉可以阻碍HMGB1、TLR4的传导;降低血清肌酶在血液中的含量;但对TLR2传导无影响。其可部分阻止免疫因子活化的程度,改善炎性细胞浸润健康肌肉组织的程度,对于皮肤、肌肉等组织的炎症导致皮肤肌肉损害起到了一定的抑制作用。天花粉对于治疗特发性炎症肌病具有一定作用。
[Abstract]:Objective: to study the effect of Trichosanthin on the signal transduction of serum creatine kinase, lactate dehydrogenase, aspartate aminotransferase and HMGB1 / TLR2 / TLR4 in rats with experimental autoimmune myositis. To investigate the repair of experimental autoimmune myositis and its mechanism after trichosanthin intervention. Materials and methods: 1. After 40 SPF SD rats were prefed in the laboratory for one week, forty prefed SD rats were randomly divided into four groups: blank group. Model group, hormone group, trichosanthin group. There were ten rats in each group. The skeletal muscle of xenogeneic animals (rabbits) is used as the raw material of muscle homogenate. The muscle homogenate and complete Freund's adjuvant were prepared and emulsified. Ten of the blank group did not need any intervention, and the remaining 30 were immunized with equal proportion muscle homogenate and complete Freund's adjuvant. The location of the injection was selected at the right and left sides of the back of the rat and subcutaneously injected. The dose of each injection was 0.35 ml, once a week. The injection was conducted on the seventh, fourteenth, 21, and 28 days after the injection. On the 35 day, the rats were injected again. After 6 times of injection, the rats were immunized. 2. 2. Drug administration observation and sample analysis were performed four weeks after immunization and after successful modeling in rats. The blank group and model group were perfused with normal saline, while the other two groups were treated by each group. The four groups were given gastric perfusion for four weeks. After four weeks, the weight of the rats was recorded in groups. After the weight was measured and anesthetized, blood was taken from the abdominal aorta as blood samples, muscle and other related tissues were taken as muscle samples, and then killed. The clinical status of the rats after drug intervention was evaluated and evaluated through the scoring rules of Lennon LA; the weight of the four groups after the last gastric perfusion was compared; and the serum creatine kinase was identified. Lactate dehydrogenase, aspartate aminotransferase and HMGB1TLR2TLR4 in rat muscle were obtained and identified by enzyme-linked immunosorbent assay (Elisa). The result is 1: 1. Trichosanthin can improve the contents of creatine kinase (CK), lactate dehydrogenase (LDH), aspartate aminotransferase (AST) in serum, and play a role in decreasing the activity of CKK LDHN AST. Trichosanth had an effect on HMGB1 TLR4 transmission, but had no effect on TLR2 transmission. Trichosanth can reduce the content of HMGB1 and TLR4 in muscle. Conclusion: Trichosanthin can block the transmission of HMGB1 and TLR4, decrease the content of serum myozyme in blood, but have no effect on TLR2 conduction. It can partly prevent the activation of immune factors, improve the degree of inflammatory cells infiltrating healthy muscle tissue, and inhibit the skin muscle damage caused by inflammation of skin, muscle and other tissues. Trichosanthin has a certain effect on the treatment of idiopathic inflammatory myopathy.
【学位授予单位】：辽宁中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R285.5
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本文编号：1803389