首页> 中文期刊> 《中国全科医学》 >肝宁方对肝细胞内质网应激相关分子GRP78蛋白及其mRNA表达的影响

肝宁方对肝细胞内质网应激相关分子GRP78蛋白及其mRNA表达的影响

摘要

Objective To investigate the effect and significance of Ganning Decoction on the mRNA and protein expression levels of GRP78 associated with hepatic endoplasmic reticulum stress(ERS)Methods This study was conducted from October 2015 to July 2016.We prepared the serum with Ganning Decoction by using the abdominal aortic blood taken from 20 intervened SPF grade healthy SD rats,and tunicamycin culture medium by adding culture medium containing fetal bovine serum to the DMSO solution with completely dissolved tunicamycin.HL-7702 hepatocytes were divided into normal control group,ERS model group,and Ganning Decoction treatment group,cultured with normal culture medium,tunicamycin culture medium,tunicamycin culture medium containing serum with Ganning Decoction,respectively.The mRNA and protein expression levels of GRP78 associated with hepatic ERS in all the groups were detected by qRT-PCR,and western blotting,respectively.Results The expression level of GRP78 mRNA in hepatocytes of ERS model group was higher than that in the normal control group(P<0.05).The expression level of GRP78 mRNA in hepatocytes of Ganning Decoction treatment group was lower than that in the ERS model group(P<0.05).The normal control group had lower expression level of GRP78 protein than the other two groups(P<0.05).Ganning Decoction treatment group had lower expression level of GRP78 protein than the ERS model group(P<0.05).Conclusion Ganning Decoction can effectively improve the impaired liver function and reduce the apoptosis of hepatocytes.The mechanism may be related to the inhibition of the expressions of GRP78 in hepatocytes.%目的 探讨肝宁方对肝细胞内质网应激(ERS)相关分子葡萄糖调节蛋白78(GRP78)蛋白及其mRNA表达的调控作用及临床意义.方法 本研究时间为2015年10月-2016年7月.SPF级SD健康大鼠20只用于制备肝宁方血清;衣霉素溶于DMSO溶液,完全溶解后加入含有胎牛血清的培养液制备衣霉素培养液.将HL-7702肝细胞分为正常对照组、ERS模型组、肝宁方治疗组.正常对照组予正常培养液培养,ERS模型组予衣霉素培养液培养,肝宁方治疗组予含肝宁方血清的衣霉素培养液培养.采用qRT-PCR测定各组肝细胞ERS中GRP78 mRNA表达水平及Western blotting法检测相应蛋白的表达水平.结果 ERS模型组肝细胞GRP78 mRNA表达水平高于正常对照组(P<0.05);肝宁方治疗组肝细胞GRP78 mRNA表达水平低于ERS模型组(P<0.05).ERS模型组与肝宁方治疗组GRP78蛋白表达水平高于正常对照组(P<0.05);肝宁方治疗组GRP78蛋白表达水平低于ERS模型组(P<0.05).结论 肝宁方能够改善肝功能损伤情况,有效降低肝细胞的凋亡,其机制可能与抑制肝细胞内GRP78的表达有关.

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