摘要
研究左归降糖解郁方调控糖尿病并发抑郁症大鼠海马组蛋白3的第18位赖氨酸(H3K18)乳酸化(H3K18la)修饰并改善星形胶质细胞乳酸生成的机制。SD大鼠按随机数字表法分为正常组,模型组,左归降糖解郁方高、中、低剂量组,灌胃给药28 d后,血糖试纸检测空腹血糖,采用ELISA法检测胰岛素并计算胰岛素抵抗指数,采用强迫游泳实验和开野实验评价动物抑郁样行为,采用RT-PCR检测钙黏蛋白1(CDH1)mRNA表达。随后将SD大鼠随机分为正常组、模型组、假手术组、L-乳酸组、左归降糖解郁方组、2-脱氧-D-葡萄糖(2-DG)组,干预结束后检测动物抑郁样行为,采用胶质纤维酸性蛋白(GFAP)/5-溴-2'-脱氧尿苷(Brdu)双重免疫荧光评估星形胶质细胞增殖水平,采用ELISA检测海马乳酸含量,采用Western blot检测细胞周期蛋白B1(cyclin B1)、H3K18la、CDH1、6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶3(PFKFB3)表达。结果表明,与正常组比较,模型组大鼠空腹血糖、胰岛素以及胰岛素抵抗水平显著升高(P<0.01),动物在强迫游泳实验中的不动时间显著延长而在开野实验中的活动次数显著减少(P<0.01)。与模型组比较,左归降糖解郁方高、中剂量可显著降低模型大鼠空腹血糖、胰岛素以及胰岛素抵抗水平(P<0.01),减少动物不动时间并增加活动次数(P<0.01)。RT-PCR实验发现,与模型组比较,左归降糖解郁方可显著降低CDH1 mRNA表达(P<0.05)。与模型组比较,左归降糖解郁方可显著上调大鼠海马乳酸含量(P<0.05),促进星形胶质细胞增殖(P<0.01),增加cyclin B1、H3K18la、PFKFB3蛋白表达(P<0.05或P<0.01),减少CDH1蛋白表达(P<0.01);2-DG可逆转左归降糖解郁方的上述作用。与假手术组比较,侧脑室注射L-乳酸可以增加海马乳酸含量及H3K18la表达(P<0.05或P<0.01),促进星形胶质细胞增殖(P<0.01),增加海马cyclin B1和PFKFB3表达而降低CDH1表达(P<0.05或P<0.01)。因此,左归降糖解郁方可增加糖尿病并发抑郁症大鼠海马PFKFB3表达并促进乳酸生成,其机制可能与增加海马H3K18la修饰并抑制CDH1表达,进而促进星形胶质细胞增殖有关。
This study aims to elucidate the underlying mechanism through which Zuogui Jiangtang Jieyu Decoction(ZGJTJYD)modulates histone 3 lysine 18 lactylation(H3K18la)modification and enhances astrocytic lactate production in the rat hippocampus with diabetes-associated depression.Sprague Dawley(SD)rats were randomly divided into a normal group,a model group,and high-,medium-,and low-dose ZGJTJYD groups.After 28 days of administration by gavage,the fasting blood glucose(FBG)was quantified by using glucometer strips.The insulin was assayed via enzyme-linked immunosorbent assay(ELISA),and the insulin resistance index(HOMA-IR)was calculated.Depressive-like behaviors were assessed by a forced swim test(FST)and an open field test(OFT).The expression of cadherin 1(CDH1)mRNA was analyzed by reverse transcription polymerase chain reaction(RT-PCR).In a subsequent independent experiment,rats were randomized into a normal group,a model group,a sham group,a L-lactate group,a ZGJTJYD group,and a 2-deoxy-D-glucose(2-DG)group.After intervention,depressive-like behaviors were assessed.Astrocyte proliferation was quantified via dual immunofluorescence staining for glial fibrillary acidic protein(GFAP)and 5-bromo-2'-deoxyuridine(Brdu),while the hippocampal lactate content was measured by ELISA.The expression of cyclin B1,H3K18la,CDH1,and 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3(PFKFB3)was analyzed by Western blot.The results showed that compared with the normal group,the model group exhibited significantly elevated FBG,insulin,and HOMA-IR(P<0.01).The immobility time in FST was significantly prolonged,and activity in OFT was significantly reduced(P<0.01).Compared with the model group,the high-and medium-dose ZGJTJYD groups exhibited significantly reduced FBG,insulin,and HOMA-IR(P<0.01),shortened immobility time,and increased activity(P<0.01).RT-PCR analysis demonstrated that compared to the model group,the ZGJTJYD groups displayed significantly downregulated expression of CDH1 mRNA(P<0.05).Compared with the model group,the ZGJTJYD groups exhibited significantly enhanced hippocampal lactate content(P<0.05),stimulated astrocyte proliferation(P<0.01),upregulated expression of cyclin B1,H3K18la,and PFKFB3 proteins(P<0.05 or P<0.01),and suppressed CDH1 expression(P<0.01).Strikingly,these effects were reversed by 2-DG.Compared with the sham group,lateral intracerebroventricular injection of L-lactate elevated the hippocampal lactate content and the H3K18la expression(P<0.05 or P<0.01),promoted astrocyte proliferation(P<0.01),increased the expression of cyclin B1 and PFKFB3,and reduced the CDH1 expression(P<0.05 or P<0.01).Therefore,ZGJTJYD increases the PFKFB3 expression and promotes lactate production in the hippocampus of rats with diabetes-associated depression.This therapeutic effect may be linked to the increased hippocampal H3K18la modification and the inhibited CDH1 expression,which in turn promotes astrocyte proliferation.
作者
熊耀
李薇
雷诗卉
王瑾茜
刘检
蔺晓源
易健
王宇红
杨蕙
XIONG Yao;LI Wei;LEI Shi-hui;WANG Jin-xi;LIU Jian;LIN Xiao-yuan;YI Jian;WANG Yu-hong;YANG Hui(the First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Hunan University of Chinese Medicine,Changsha 410208,China)
出处
《中国中药杂志》
北大核心
2025年第22期6410-6418,共9页
China Journal of Chinese Materia Medica
基金
国家自然科学基金面上项目(82474476)
湖南省重大基础研究项目(2025JC0006)
湖南省自然科学基金项目(2024JJ4033,2023JJ30476)
湖南省教育厅优秀青年项目(24B0374)
长沙市自然科学基金项目(kq2403108)。
