切换至 "中华医学电子期刊资源库"
高级检索
中华妇幼临床医学杂志(电子版)

中华妇幼临床医学杂志(电子版) ›› 2022, Vol. 18 ›› Issue (05) : 528 -539. doi: 10.3877/cma.j.issn.1673-5250.2022.05.006

论著

cblC型甲基丙二酸血症MMACHC基因新突变对小鼠神经细胞凋亡及Wnt/β-catenin信号通路的作用机制
周伟1, 蔡恒2, 范海迪3, 李惠中1, 王传霞1, 顾茂胜1,†()   
  1. 1徐州市妇幼保健院遗传医学中心，徐州　221009
    2徐州医科大学，徐州　221004
    3淮安市第一人民医院第一分院检验科，淮安　223300
  • 收稿日期:2021-09-24 修回日期:2022-04-29 出版日期:2022-10-01
  • 通信作者: 顾茂胜

Mechanism of MMACHC gene mutation of cblC-type methylmalonic acidemia on apoptosis and Wnt/β-catenin signaling pathway in mouse neural cells

Wei Zhou1, Heng Cai2, Haidi Fan3, Huizhong Li1, Chuanxia Wang1, Maosheng Gu1,†()   

  1. 1Center of Genetic Medicine, Xuzhou Maternity and Child Health Care Hospital, Xuzhou 221009, Jiangsu Province, China
    2Xuzhou Medical University, Xuzhou 221004, Jiangsu Province, China
    3Clinical Laboratory, The First Branch of Huai′an First Poeple′s Hospital, Huai′an 223300, Jiangsu Province, China
  • Received:2021-09-24 Revised:2022-04-29 Published:2022-10-01
  • Corresponding author: Maosheng Gu
  • Supported by:
    Maternal and Child Health Research Project of Jiangsu Province(F201912); Xuzhou Science and Technology Program(KC19028, KC20074)
全文 ( ) 下载PDF ( ) 导出引用
引用本文：

周伟, 蔡恒, 范海迪, 李惠中, 王传霞, 顾茂胜. cblC型甲基丙二酸血症MMACHC基因新突变对小鼠神经细胞凋亡及Wnt/β-catenin信号通路的作用机制[J]. 中华妇幼临床医学杂志(电子版), 2022, 18(05): 528-539.

Wei Zhou, Heng Cai, Haidi Fan, Huizhong Li, Chuanxia Wang, Maosheng Gu. Mechanism of MMACHC gene mutation of cblC-type methylmalonic acidemia on apoptosis and Wnt/β-catenin signaling pathway in mouse neural cells[J]. Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition), 2022, 18(05): 528-539.

目的

探讨cblC型甲基丙二酸血症(MMA)致病基因MMACHC新突变，对小鼠神经细胞凋亡及Wnt/β-catenin信号通路的作用机制。

方法

选择2017年5月和7月，江苏省新生儿疾病筛查中心徐州分中心筛查确诊2例cblC型MMA患儿的血液样本中，检出的c.626_627del及c.228_231del 2种MMACHC基因新错义突变为研究材料。分析MMACHC基因的2种新突变所累及的氨基酸残基的保守性及预测其致病性后，构建MMACHC基因2种新突变重组质粒，并与野生型MMACHC基因重组质粒，分别瞬时转染小鼠神经细胞(神经元细胞株HT22及神经干细胞株C17.2)进行体外实验。将MMACHC基因c.626_627del质粒转染的小鼠HT22、C17.2细胞，分别纳入神经元观察1组、神经干观察1组；将MMACHC基因c.228_231del质粒转染的小鼠HT22、C17.2细胞，分别纳入神经元观察2组、神经干观察2组；将野生型MMACHC基因重组质粒转染的小鼠HT22、C17.2细胞，分别纳入神经元对照组、神经干对照组。采用免疫荧光染色、TUNEL荧光检测及蛋白质印迹法(Western blotting)等，分析小鼠神经细胞凋亡情况及Wnt/β-catenin信号通路相关蛋白表达情况。采用单因素方差分析及Dunnett t检验，对2个观察组分别与对照组神经细胞的细胞凋亡率，凋亡相关蛋白及Wnt/β-catenin信号通路相关蛋白表达水平进行统计学比较。本研究经过徐州妇幼保健院伦理委员会批准[审批文号：〔2019〕伦审第(09)号]。

结果

①MMACHC基因2种新突变所在氨基酸位点(第77位及第209位)，在人源、家鼠、褐鼠、斑马鱼及猕猴5个物种间高度保守。生物信息学分析显示，2种突变均可能具有致病性。②与对照组相比，与之对应的2个观察组神经细胞的细胞核DAPI染色着色更深(亮蓝色)，TUNEL荧光阳性(绿色)细胞数更多。神经元观察1组细胞凋亡率(DAPI染色)及凋亡率(TUNEL染色)分别为(25.8±1.1)%、(23.1±1.4)%，均高于神经元对照组的(4.9±0.9)%、(3.2±0.4)%，并且差异均有统计学意义(t＝25.95、24.41，均为P<0.001)；与此具有相似结果的是，神经元观察2组与神经元对照组比较，以及神经干观察1组、2组分别与神经干对照组比较，差异亦均有统计学意义(均为P<0.05)。③神经元观察1组、2组HT22细胞中凋亡相关蛋白caspase-3、cleaved caspase-3及细胞质内β-连环蛋白(β-catenin)表达水平，均分别高于神经元对照组，而Wnt/β-catenin信号通路相关蛋白突触后致密物(PSD)-95、myc-MMACHC、磷酸化-糖原合成酶激酶(p-GSK)-3β(Ser)及细胞核内β-catenin表达水平，则均分别低于神经元对照组，并且差异均有统计学意义(P<0.05)。④神经干观察1组C17.2细胞中caspase-3、PSD-95、myc-MMACHC、p-GSK-3β(Ser)及细胞核内β-catenin表达水平，均低于神经干对照组，而cleaved caspase-3、GSK-3β及细胞质内β-catenin表达水平，则均高于神经干对照组，并且上述差异均有统计学意义(P<0.05)。神经干观察2组C17.2细胞中caspase-3、cleaved caspase-3、GSK-3β及细胞质内β-catenin表达水平，均高于神经干对照组，而细胞核内β-catenin表达水平，则低于神经干对照组，并且上述差异均有统计学意义(P<0.05)。

结论

MMACHC基因2种新突变可能通过抑制GSK-3β-Wnt/β-catenin信号通路，而促进小鼠神经细胞凋亡。

关键词: 甲基丙二酸血症, MMACHC基因, 神经元, 神经干细胞, 细胞凋亡, 糖原合成酶激酶3, β连环素, 小鼠
Objective

To investigate the mechanism of novel variants in MMACHC gene resulting in cblC-type methylmalonic acidemia (MMA) on neural cell apoptosis and Wnt/β-catenin signaling pathway in mice.

Methods

Two novel missense variants of c. 626_627del and C. 228_231del of MMACHC gene which were detected in blood samples of two infants with cblC-type MMA screening and diagnosed by Xuzhou Branch Center of Jiangsu Newborn Screening Center in May and July 2017, were selected as research materials. After analyzing the conservation of amino acid residues involved in the two novel variants and predicting the pathogenicity, two new recombinant plasmids of MMACHC gene were constructed, and with the recombinant plasmids of wild-type MMACHC gene, they were transiently transfected into mouse nerve cells (neuron cell line HT22 and nerve stem cell line C17.2) for in vitro experiments. Mice HT22 and C17.2 cells transfected with MMACHC gene c.626_627del plasmid were included into neuron observation group 1 and nerve stem observation group 1, respectively. Mice HT22 and C17.2 cells transfected with MMACHC gene C. 228_231del plasmid were included into neuron observation group 2 and nerve stem observation group 2, respectively. Mice HT22 and C17.2 cells transfected with wild-type MMACHC gene recombinant plasmid were included into neuron control group and nerve stem control group, respectively. Immunofluorescence staining, TUNEL fluorescence detection and Western blotting were used to analyze the apoptosis of mouse nerve cells and the expression of Wnt/β-catenin signaling pathway related proteins. One-way ANOVA and Dunnett t test were used to statistically compare the apoptosis rate, apoptosis-related proteins and the expression levels of Wnt/β-catenin signaling pathway related proteins between two observation groups and control group, respectively. This study was approved by the Ethics Committee of Xuzhou Maternity and Child Health Care Hospital [Approval No.2019(09)].

Results

① The amino acid sites of two novel variants in MMACHC gene (positions 77 and 209) were highly conserved among H. sapiens, M. musculus, R. norvegicus, D. rerioand and M. mulatta. Bioinformatics analysis indicated that both variants may be pathogenic. ② Compared to control group, the DAPI staining of nuclei of nerve cells in corresponding two observation groups were darker (bright blue), and the number of cells with positive TUNEL fluorescence (green) were more. The apoptosis rate (DAPI staining) and apoptosis rate (TUNEL staining) of neuron observation group 1 were (25.8±1.1)% and (23.1±1.4)%, respectively, which were significantly higher than those of (4.9±0.9)% and (3.2±0.4)% in neuron control group, and the differences were statistically significant (t=25.95, 24.41; all P<0.001). With similar results to this were, neuron observation group 2 compared with neuron control group, also nerve stem observation group 1 and group 2 compared with nerve stem control group, respectively, and the differences were statistically significant (all P<0.05). ③ The expression levels of apoptosis-related proteins of caspase-3 and cleaved caspase-3, and intracytoplasmic β-catenin in HT22 cells in neuron observation groups 1 and 2 were higher than those in neuron control group, respectively; while the expression levels of Wnt/β-catenin signaling pathway related proteins of postsynaptic density(PSD)-95, myc-MMACHC, phosphorylation-glycogen synthase kinase(p-GSK)-3β(Ser) and nuclear nucleus β-catenin were lower than those of neuron control group, respectively. And the above differences were statistically significant (P<0.05). ④ The expression levels of caspase-3, PSD-95, myc-MMACHC, p-GSK-3β(Ser) and nuclear nucleus β-catenin in C17.2 cells in nerve stem observation group 1 were lower than those in nerve stem control group; while the expression levels of cleaved caspase-3, GSK-3β and intracytoplasmic β-catenin were significantly higher than those in nerve stem control group. And the above differences were statistically significant (P<0.05). The expression levels of caspase-3, cleaved caspase-3, GSK-3β and intracytoplasmic β-catenin of C17.2 cells in nerve stem observation group 2 were higher than those in nerve stem control group, while the expression level of nuclear nucleus β-catenin was lower than that in nerve stem control group. And the above differences were statistically significant (P<0.05).

Conclusions

Two novel variants in MMACHC gene might inhibit GSK-3β-Wnt/β-catenin signaling pathway to promote nerve cell apoptosis in mice.

Key words: Methylmalonic academia, MMACHC gene, Neurons, Neural stem cells, Apoptosis, Glycogen synthase kinase 3, Beta catenin, Mice
图1 对人源、家鼠、褐鼠、斑马鱼及猕猴5个不同物种MMACHC基因2种新突变的氨基酸位点同源性分析(图1A表示c.228_231del突变的MMACHC蛋白氨基酸位点同源性分析；图1B表示c.626_627del突变的MMACHC蛋白氨基酸位点同源性分析)注：Homo sapiens为人类、Mus musculus为家鼠、Rattus norvegicus为褐鼠、Danio rerio为斑马鱼、Macaca mulatta为猕猴
图2 对人源MMACHC蛋白氨基酸残基空间位阻示意图[图2A：MMACHC蛋白野生型与p.D77Qfs*22突变型组合的三维结构示意图；图2B：MMACHC蛋白野生型第77位天冬氨酸残基空间结构(绿色)示意图；图2C：MMACHC蛋白突变型第77位谷氨酸残基空间结构(红色)示意图；图2D：MMACHC蛋白野生型与突变型第77位氨基酸残基空间结构组合示意图；图2E：MMACHC蛋白野生型与p.V209Dfs*35突变型组合的三维结构示意图；图2F：MMACHC蛋白野生型第209位缬氨酸残基空间结构(绿色)示意图；图2G：MMACHC蛋白突变型第209位天冬氨酸残基空间结构(红色)示意图；图2H：MMACHC蛋白野生型与突变型第209位氨基酸残基空间结构组合示意图]注：Asp为天冬氨酸，Gln为谷氨酰胺，Val为缬氨酸
图3 人源MMACHC蛋白三维结构及其表面电荷分布示意图(图3A：MMACHC蛋白野生型及2种突变型组合的三维结构示意图，绿色为野生型、红色为p.D77Qfs*22突变型、灰蓝色为p.V209Dfs*35突变型；图3B：MMACHC蛋白结构域简图，并标注2种突变的氨基酸位置；图3C~3E：MMACHC蛋白3种型表面电荷分布示意图，蓝色为负电荷聚集区、红色为正电荷聚集区)
表1 转染MMACHC重组质粒后，3组小鼠神经元HT22细胞凋亡率及凋亡相关蛋白表达水平比较(±s)
组别 例数 细胞凋亡率(%)(DAPI染色) caspase-3相对表达量 cleaved caspase-3相对表达量 细胞凋亡率(%)(TUNEL染色)
①神经元观察1组 3 25.8±1.1 0.86±0.01 0.80±0.00 23.1±1.4
②神经元观察2组 3 12.5±1.2 1.10±0.03 1.13±0.02 32.5±1.2
③神经元对照组 3 4.9±0.9 0.65±0.00 0.69±0.02 3.2±0.4
总体比较(F值/P值)   289.40/<0.001 394.30/<0.001 720.70/<0.001 572.50/<0.001
两两比较(t值/P值)          
  ① vs ②   14.04/0.001 12.06/<0.001 35.87/<0.001 8.91/0.001
  ① vs ③   25.95/<0.001 32.40/<0.001 9.60/0.001 24.41/<0.001
  ② vs ③   8.66/0.001 24.31/<0.001 29.85/<0.001 38.86/<0.001
表2 转染MMACHC重组质粒后，3组小鼠神经干C17.2细胞凋亡率及凋亡相关蛋白表达水平比较(±s)
组别 例数 凋亡率(%)(DAPI染色) caspase-3相对表达量 cleaved caspase-3相对表达量 凋亡率(%)(TUNEL染色)
①神经干观察1组 3 45.80±1.07 1.19±0.01 1.45±0.12 18.9±1.0
②神经干观察2组 3 35.53±3.11 1.40±0.05 1.40±0.08 22.1±1.0
③神经干对照组 3 5.70±0.26 1.23±0.01 0.74±0.02 2.2±0.5
总体比较(F值/P值)   358.00/<0.001 41.30/<0.001 71.40/<0.001 436.40/<0.001
两两比较(t值/P值)          
  ① vs ②   5.40/0.006 7.32/0.002 0.60/0.584 3.72/0.204
  ① vs ③   63.02/<0.001 4.25/0.013 10.32/<0.001 30.60/<0.001
  ② vs ③   16.54/<0.001 5.69/0.005 14.92/<0.001 25.51/<0.001
图4 转染MMACHC突变质粒后小鼠神经细胞凋亡变化[图4A：3组HT22细胞的免疫荧光图(细胞核DAPI荧光染色呈蓝色，高倍镜)；图4B：Western blotting检测3组HT22细胞凋亡相关蛋白相对表达量；图4C: 3组HT22细胞的免疫荧光图(TUNEL荧光染色的凋亡细胞呈绿色，高倍镜)；图4D：3组C17.2细胞的免疫荧光图(细胞核DAPI荧光染色呈蓝色，高倍镜)图4E：Western blotting检测3组C17.2细胞凋亡相关蛋白相对表达量；图4F: 3组C17.2细胞的免疫荧光图(TUNEL荧光染色的凋亡细胞呈绿色，高倍镜)]注：1表示MMACHC基因c.626_627del质粒转染神经细胞，2表示MMACHC基因c.228_231del质粒转染神经细胞，3表示野生型MMACHC基因重组质粒转染神经细胞。α-tubulin为α-微管蛋白(内参蛋白)，caspase为半胱氨酸天冬氨酸蛋白酶，cleaved caspase为活化的半胱氨酸天冬氨酸蛋白酶。Western blotting为蛋白质印迹法
表3 3组小鼠神经元H2TT细胞Wnt/β-catenin信号通路相关蛋白表达水平比较(±s)
组别 例数 PSD-95相对表达量 myc-MMACHC相对表达量 p-GSK-3β(Ser9)相对表达量 GSK-3β相对表达量 β-catenin相对表达量
细胞质内 细胞核内
①神经元观察1组 3 0.85±0.00 0.78±0.00 0.89±0.00 0.87±0.01 1.36±0.02 0.49±0.02
②神经元观察2组 3 0.96±0.02 0.88±0.01 0.95±0.02 0.87±0.02 1.12±0.01 0.68±0.01
③神经元对照组 3 1.02±0.01 1.24±0.01 1.04±0.02 0.90±0.03 1.10±0.00 0.76±0.00
总体比较(F值/P值)   119.70/<0.001 1 211.00/<0.001 72.29/<0.001 1.33/0.033 357.20/<0.001 537.10/<0.001
两两比较(t值/P值)              
  ① vs ②   8.33/0.001 11.68/<0.001 4.51/0.011 0.17/0.871 18.45/<0.001 19.15/<0.001
  ① vs ③   44.73/<0.001 53.16/<0.001 15.91/<0.001 1.59/0.188 20.82/<0.001 27.19/<0.001
  ② vs ③   4.41/0.012 30.25/<0.001 6.16/0.004 1.53/0.199 4.02/0.016 19.05/<0.001
图6 MMACHC基因2种突变质粒转染小鼠神经干细胞C17.2后，Wnt/β-catenin信号通路相关蛋白Western blotting电泳图
表4 3组小鼠神经干C17.2细胞Wnt/β-catenin信号通路相关蛋白表达水平比较(±s)
组别 例数 PSD-95相对表达量 myc-MMACHC相对表达量 p-GSK-3β(Ser9)相对表达量 GSK-3β相对表达量 β-catenin相对表达量
细胞质内 细胞核内
①神经干观察1组 3 1.17±0.02 0.92±0.01 1.76±0.01 1.82±0.00 0.92±0.02 0.91±0.01
②神经干观察2组 3 1.23±0.01 1.24±0.01 1.83±0.00 1.84±0.00 0.86±0.01 0.96±0.01
③神经干对照组 3 1.29±0.04 1.25±0.02 1.83±0.01 1.68±0.02 0.66±0.01 1.01±0.01
总体比较(F值/P值)   15.53/0.004 573.90/<0.001 138.50/<0.001 172.80/<0.001 370.50/<0.001 99.80/<0.001
两两比较(t值/P值)              
  ① vs ②   5.53/0.005 32.50/<0.001 15.91/<0.001 5.41/0.006 5.38/0.006 6.48/0.003
  ① vs ③   4.66/0.010 27.40/<0.001 12.80/<0.001 12.55/<0.001 23.34/<0.001 21.13/<0.001
  ② vs ③   2.21/0.091 1.08/0.339 1.02/0.365 14.28/<0.001 25.19/<0.001 6.02/0.004
[1]
Rashka C, Hergalant S, Dreumont N, et al. Analysis of fibroblasts from patients with cblC and cblG genetic defects of cobalamin metabolism reveals global dysregulation of alternative splicing[J]. Hum Mol Genet, 2020, 29(12): 1969-1985. DOI: 10.1093/hmg/ddaa027.
[2]
Liu J, Peng Y, Zhou N, et al. Combined methylmalonic acidemia and homocysteinemia presenting predominantly with late-onset diffuse lung disease: a case series of four patients[J]. Orphanet J Rare Dis, 2017, 12(1): 58. DOI: 10.1186/s13023-017-0610-8.
[3]
Wongkittichote P, Cunningham G, Summar ML, et al. Tricarboxylic acid cycle enzyme activities in a mouse model of methylmalonic aciduria[J]. Mol Genet Metab, 2019, 128(4): 444-451. DOI: 10.1016/j.ymgme.2019.10.007.
[4]
Ripp C, Loth J, Petrova I, et al. Drosophila Ror is a nervous system-specific co-receptor for Wnt ligands[J]. Biol Open, 2018, 7(11): bio033001. DOI: 10.1242/bio.033001.
[5]
Nusse R, Clevers H. Wnt/β-catenin signaling, disease, and emerging therapeutic modalities[J]. Cell, 2017, 169(6): 985-999. DOI: 10.1016/j.cell.2017.05.016.
[6]
Perugorria MJ, Olaizola P, Labiano I, et al. Wnt-β-catenin signalling in liver development, health and disease[J]. Nat Rev Gastroenterol Hepatol, 2019, 16(2): 121-136. DOI: 10.1038/s41575-018-0075-9.
[7]
韩连书，黄倬，叶军，等. 甲基丙二酰辅酶A变位酶基因新错义突变功能研究[C]//第十四次全国医学遗传学学术会议论文集，南宁，2015. 北京：中国遗传学会人类与医学遗传专业委员会，2015: 97.
[8]
吕书博，张展，赵德华，等. JNK/MAPK信号通路介导甲基丙二酸血症脑损伤的机制研究[J]. 东南大学学报(医学版), 2019, 38(5): 837-842. DOI: 10.3969/j.issn.1671-6264.2019.05.013.
[9]
顾学范．临床遗传代谢病[M]. 北京：人民卫生出版社，2015: 106-107.
[10]
Zhou W, Li H, Wang C, et al. Newborn screening for methylmalonic acidemia in a Chinese population: molecular genetic confirmation and genotype phenotype correlations[J]. Front Genet, 2019, 9: 726. DOI: 10.3389/fgene.2018.00726.
[11]
Ktena YP, Ramstad T, Baker EH, et al. Propofol administration in patients with methylmalonic acidemia and intracellular cobalamin metabolism disorders: a review of theoretical concerns and clinical experiences in 28 patients[J]. J Inherit Metab Dis, 2015, 38(5): 847-853. DOI: 10.1007/s10545-015-9816-x.
[12]
王秀利，顾茂胜，周伟，等. 新生儿甲基丙二酸血症的筛查与基因突变分析[J]. 中华围产医学杂志，2018, 21(8): 541-550. DOI: 10.3760/cma.j.issn.1007-9408.2018.08.007.
[13]
Schunk SJ, Floege J, Fliser D, et al. WNT-β-catenin signalling - a versatile player in kidney injury and repair[J]. Nat Rev Nephrol, 2021, 17(3): 172-184. DOI: 10.1038/s41581-020-00343-w.
[14]
Anthony CC, Robbins DJ, Ahmed Y, et al. Nuclear regulation of Wnt/β-catenin signaling: it′s a complex situation[J]. Genes (Basel), 2020, 11(8): 886. DOI: 10.3390/genes11080886.
[15]
Wang C, Li D, Cai F, et al. Mutation spectrum of MMACHC in Chinese pediatric patients with cobalamin C disease: a case series and literature review[J]. Eur J Med Genet, 2019, 62(10): 103713. DOI: 10.1016/j.ejmg.2019.103713.
[16]
Mascarenhas R, Li Z, Gherasim C, et al. The human B12 trafficking protein CblC processes nitrocobalamin[J]. J Biol Chem, 2020, 295(28): 9630-9640. DOI: 10.1074/jbc.RA120.014094.
[17]
马飞，石聪聪，梁普平，等. 利用CRISPR/Cas9构建甲基丙二酸血症cblC型W203X突变小鼠模型[J]. 中国当代儿科杂志，2019, 21(8): 824-829. DOI: 10.7499/j.issn.1008-8830.2019.08.016.
[18]
Fernando WMADB, Martins IJ, Morici M, et al. Sodium butyrate reduces brain amyloid-β levels and improves cognitive memory performance in an Alzheimer′s disease transgenic mouse model at an early disease stage[J]. J Alzheimers Dis, 2020, 74(1): 91-99. DOI: 10.3233/JAD-190120.
[19]
Chen CS, Tseng YT, Hsu YY, et al. Nrf2-Keap1 antioxidant defense and cell survival signaling are upregulated by 17β-estradiol in homocysteine-treated dopaminergic SH-SY5Y cells[J]. Neuroendocrinology, 2013, 97(3): 232-241. DOI: 10.1159/000342692.
[20]
Yu L, Chen Y, Wang W, et al. Multi-vitamin B supplementation reverses hypoxia-induced tau hyperphosphorylation and improves memory function in adult mice[J]. J Alzheimers Dis, 2016, 54(1): 297-306. DOI: 10.3233/JAD-160329.
[21]
刁丽梅，李华琼，张庆梅，等. 定痫丸对急性期癫痫小鼠海马组织Wnt/β-Catenin信号通路相关蛋白及神经元凋亡的影响[J]. 中华中医药学刊，2020, 38(3): 45-49,后插4. DOI: 10.13193/j.issn.1673-7717.2020.03.012.
[1] 孔莹莹, 谢璐涛, 卢晓驰, 徐杰丰, 周光居, 张茂. 丁酸钠对猪心脏骤停复苏后心脑损伤的保护作用及机制研究[J]. 中华危重症医学杂志(电子版), 2023, 16(05): 355-362.
[2] 张晓燕, 肖东琼, 高沪, 陈琳, 唐发娟, 李熙鸿. 转录因子12过表达对脓毒症相关性脑病大鼠大脑皮质的保护作用及其机制[J]. 中华妇幼临床医学杂志(电子版), 2023, 19(05): 540-549.
[3] 刘硕儒, 王功炜, 张斌, 李书豪, 胡成. 新型溶瘤病毒M1激活内质网应激致前列腺癌细胞凋亡的机制[J]. 中华腔镜泌尿外科杂志(电子版), 2023, 17(04): 388-393.
[4] 唐英俊, 李华娟, 王赛妮, 徐旺, 刘峰, 李羲, 郝新宝, 黄华萍. 人脐带间充质干细胞治疗COPD小鼠及机制分析[J]. 中华肺部疾病杂志(电子版), 2023, 16(04): 476-480.
[5] 邓春文, 陈嵩, 钟裴, 闵师强, 万健. LncRNA CRNDE通过miR-181a-5p/SOX6轴调节脂多糖诱导人肺泡上皮细胞的炎症反应和细胞凋亡[J]. 中华细胞与干细胞杂志(电子版), 2023, 13(03): 129-136.
[6] 张师垚, 徐岩岩, 张琦, 李春强, 赵智成, 刘刚. m6A结合蛋白YTHDC2调节p38MAPK信号通路影响结直肠癌细胞凋亡[J]. 中华结直肠疾病电子杂志, 2023, 12(02): 117-124.
[7] 李青霖, 宋仁杰, 周飞虎. 一种重型劳力性热射病相关急性肾损伤小鼠模型的建立与探讨[J]. 中华肾病研究电子杂志, 2023, 12(05): 265-270.
[8] 朱泽超, 杨新宇, 李侑埕, 潘鹏宇, 梁国标. 染料木黄酮通过SIRT1/p53信号通路对蛛网膜下腔出血后早期脑损伤的作用[J]. 中华神经创伤外科电子杂志, 2023, 09(05): 261-269.
[9] 于迪, 于海波, 吴焕成, 李玉明, 苏彬, 陈馨. 发状分裂相关增强子1差异表达对胆固醇刺激下血管内皮细胞的影响[J]. 中华脑科疾病与康复杂志(电子版), 2023, 13(05): 264-270.
[10] 王淑友, 宋晓晶, 贾术永, 王广军, 张维波. 肝脏去唾液酸糖蛋白受体靶向活体荧光成像评估酒精性肝损伤肝脏功能的研究[J]. 中华消化病与影像杂志(电子版), 2023, 13(06): 443-446.
[11] 李秋琼, 薛静, 王敏, 陈芬, 肖美芳. NSE、SIL-2R、TNF-α检测对小儿病毒性脑膜炎与细菌性脑膜炎的诊断价值[J]. 中华临床医师杂志(电子版), 2023, 17(03): 303-307.
[12] 殷秀梅, 杨丽红, 姜涛, 杜元灏. 基于神经干细胞探讨巢蛋白在缺血性脑卒中中的作用机制及针刺效应[J]. 中华针灸电子杂志, 2023, 12(03): 111-116.
[13] 梁伟, 王晓彬, 洪笑阳, 蔡明岳, 梁礼聪, 陈烨, 黄培凯, 刘铭宇, 林立腾, 朱康顺. 原位肝癌小鼠微波消融术后复发模型的构建[J]. 中华介入放射学电子杂志, 2023, 11(02): 133-139.
[14] 买买提·依斯热依力, 王永康, 阿巴伯克力·乌斯曼, 克力木·阿不都热依木. 基于16s rRNA测序分析小鼠高脂饮食诱导肥胖的肠道菌群结构特征[J]. 中华肥胖与代谢病电子杂志, 2023, 09(01): 12-16.
[15] 邱甜, 杨苗娟, 胡波, 郭毅, 何奕涛. 亚低温治疗脑梗死机制的研究进展[J]. 中华脑血管病杂志(电子版), 2023, 17(05): 518-521.
阅读次数
全文


摘要

版权所有 中华医学会 中华医学电子音像出版社有限责任公司  京ICP备14006079号-1  网络出版服务许可证：(署)网出证(京)字第075号