心不藏神不寐大鼠海马与心肌组织中凋亡相关蛋白表达及细胞凋亡情况研究  

Expression of apoptosis-related proteins and apoptosis in hippocampus and myocardium of rats with insomnia due to heart not storing spirit

作　　者：胡龙涛 张星平 陈旭 王菲[3] 邓汉沣 HU Longtao;ZHANG Xingping;CHEN Xu;Wang Fei;Deng Hanfeng(Fourth Clinical Medical College of Xinjiang Medical University,Urumqi 830099,Xinjiang,China;Traditional Chinese Medicine Hospital Affiliated of Xinjiang Medical University,Urumqi 830099,Xinjiang,China;Shaanxi University of Chinese Medicine,Xianyang 712000,Shaanxi,China)

机构地区：[1]新疆医科大学第四临床医学院,新疆乌鲁木齐830099 [2]新疆医科大学附属中医医院,新疆乌鲁木齐830099 [3]陕西中医药大学,陕西咸阳712000

出　　处：《现代中西医结合杂志》2024年第15期2083-2087,共5页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家自然科学基金资助项目(82160873,81960837,81560762,81260526);新疆自然科学基金重点项目(2022D01D48);全国名老中医药专家传承工作室(国中医药人教函[2022]75号);国家中管局高水平中医药重点学科建设项目(国中医药人教函[2023]85号);新疆“十四五”中医学重点学科(新教函[2022]112号)。

摘　　要：目的探讨心不藏神不寐与海马和心肌组织细胞凋亡的关系。方法将14只SPF级雄性SD大鼠随机分为模型组与正常组,每组7只。模型组大鼠采用咖啡因+对氯苯丙氨酸腹腔注射方法构建心不藏神不寐模型,正常组大鼠给予同等剂量生理盐水腹腔注射。注射10 d后处死2组大鼠,RT-PCR法检测海马和心肌组织中B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)和半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)mRNA表达情况;免疫组化染色观察海马和心肌组织中Bcl-2、Bax、Caspase-3蛋白阳性表达情况;TUNEL染色观察海马与心肌组织细胞情况,并计算凋亡率。结果模型组大鼠海马组织中Bax、Caspase-3 mRNA相对表达量和心肌组织中Caspase-3 mRNA相对表达量均明显高于正常组(P均<0.05),海马组织和心肌组织中Bcl-2 mRNA相对表达量均明显低于正常组(P<0.05);模型组大鼠海马和心肌组织细胞均出现凋亡,细胞凋亡率均明显高于正常组(P均<0.05)。结论心不藏神不寐大鼠海马与心肌组织细胞发生明显凋亡,心不藏神不寐的发生发展与海马和心肌组织细胞的过度凋亡有关。Objective It is to explore the relationship between insomnia due to heart not storing spirit and apoptosis in hippocampus and myocardium.Methods Fourteen SPF male SD rats were randomly divided into model group and normal group,with 7 rats in each group.The rats in the model group were given caffeine+p-chlorophenylalanine(PCPA)by intraperitoneal injection to establish models of insomnia due to heart not storing spirit,and the rats in the normal group were given the same dose of normal saline by intraperitoneal injection.After 10 days of injection,the rats in the 2 groups were executed,and the mRNA expressions of B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),and cysteine aspartate protease-3(Caspase-3)in the hippocampus and myocardial tissues were detected by RT-PCR;the positive expressions of Bcl-2,Bax,and Caspase-3 proteins in the hippocampus and myocardial tissues were observed by immunohistochemical staining;the cells in the hippocampus and myocardial tissues were observed by TUNEL staining,and the apoptosis rate was calculated.Results The relative expressions of Bax and Caspase-3 mRNA in hippocampal tissue and Caspase-3 mRNA in myocardial tissue of the rats in the model group were significantly higher than those in the normal group(all P<0.05),and the relative expressions of Bcl-2 mRNA in hippocampal and myocardial tissues were significantly lower than those in the normal group(all P<0.05).Apoptosis was observed in both hippocampal and myocardial tissue cells of rats in the model group,and the rates of apoptosis were all significantly higher than those in the normal group(both P<0.05).Conclusion Significant apoptosis occurred in the hippocampus and myocardial tissue of rats with insomnia due to heart not storing spirit,and the development of insomnia due to heart not storing spirit was associated with excessive apoptosis of the hippocampus and myocardial tissue.

关 键 词：不寐 心不藏神 大鼠 海马组织 心肌组织 凋亡 

分 类 号：R256.23[医药卫生—中医内科学]