中草药方剂对鲫鱼热应激蛋白HSP 70表达的影响

Effects of the Chinese Herbal Compounds on the Heat Shock Protein 70 Expression of Crucian Carp

选用4种中草药方剂AH、CD、Ⅳ和APS,每日按体质量的3%混饲鲫鱼30 d。采用实时荧光定量RT-PCR方法,监测适温(18℃)及热刺激(28℃)1 h后的鲫鱼脑及肝脏中HSP 70表达水平变化。试验结果表明,对照组鲫鱼体内2种组织中HSP 70的表达量在热刺激条件下均有不同程度的升高,且适温和热应激条件下,鲫鱼肝脏组织中HSP 70表达量均明显高于脑组织。而AH组、IV组和APS组热刺激后HSP 70 mRNA含量升高,但CD组热刺激后HSP 70 mRNA转录水平降低,且在肝脏中表达水平降低极为显著。4种方剂中尤以补血养气为主要功能的AH方剂在鲫鱼肝脏中对HSP 70表达水平有极大的促进作用,在适温及热应激条件下,分别是对照组的4.86,3.43倍。因此,中草药方剂AH对促进鲫鱼HSP 70基因高水平表达有较好的作用,有助于鱼体抵抗热刺激防止热损伤,可作为有效的鲫鱼抗热应激备选药物。

In this study,real time quantitative PCR method was performed to understand the effects of four kinds of Chinese herbal compounds,including AH,CD,Ⅳ and APS on the expression of heat shock protein 70 in the liver and brain tissues of Crucian carp.A total of 60 fishes were divided into 5 groups and fed on diet with 3% Chinese herbal compounds of body weight.After feeding for 30 days continuously,the expression of HSP 70 in the liver and brain tissues was investigated both under the optimum temperature（18℃） and heat shock temperature（28℃） for an hour.The results indicated that the expression of HSP 70 in control group increased in varying degree during the heat shock stress,and it was significantly higher in the liver than that of brain tissues both under the normal condition or heat shock stress.Of four experimental groups,the content of HSP 70 mRNA was found to increase in AH,IV and APS groups while decrease in CD group after heat shock stress,significantly decreasing in the liver tissues.In addition,AH Chinese herb,with the main function of replenishing blood and Qi,could greatly enhance the expression of HSP 70 in the liver of Crucian carp by 4.86 folds and 3.43 folds compared to control group under 18℃ and 28℃,respectively.In conclusion,AH compound was found to promote high-level expression of HSP 70 gene in Crucian carp,to be helpful to resist heat shock and could be applied as an effective anti-heat stress drug.