膳食亚麻木酚素通过调控Bcl2/Bax/caspase3信号轴通道改善大鼠母体反式脂肪酸暴露所导致的子代肾损伤

doi:10.12122/j.issn.1673-4254.2025.12.13

摘要/Abstract

摘要：

目的探究亚麻木酚素（SDG）对妊娠期及哺乳期母鼠反式脂肪酸（TFA）暴露致子鼠肾脏损伤的保护作用及潜在作用机制。方法 30只C57BL/6雌鼠随机分为5组，分别为对照组、TFA暴露组、SDG低（10 mg/kg）、中（20 mg/kg）、高（30 mg/kg）干预组，6只/组。采用脲酶法和肌氨酸氧化酶法分别检测子鼠血肌酐（BUN）和血尿素氮（CRE）的水平；通过网络药理学初步探究SDG对TFA暴露致肾损伤的潜在保护机制，分子对接初步探究了SDG与Bcl2/Bax/caspase-3的结合能力。在此基础上选择凋亡相关的Bcl2/Bax/caspase-3信号轴对实验小鼠进行后续分析；蛋白免疫印迹法检测cleaved-caspase-3、Bax、Bcl2蛋白水平。结果动物实验发现，TFA组子鼠血清中尿素氮及肌酐水平高于对照组（P<0.05），而干预后SDG中、高剂量组子鼠血清中的尿素氮及肌酐较TFA暴露组降低（P<0.05）。网络药理学和分子对接结果提示，SDG可能通过作用凋亡相关的Bcl2/Bax/caspase-3轴改善肾损伤。进一步实验结果显示：TFA暴露组较对照组子鼠肾组织阳性凋亡细胞增多、凋亡指数上升；cleaved-caspase-3蛋白水平升高及Bcl2/Bax蛋白水平比值下降（P<0.05）。与TFA暴露组相比，低、中、高剂量SDG干预组凋亡指数下降、cleaved-caspase-3蛋白水平下降、Bcl2/Bax蛋白水平比值上升（P<0.05）。结论母鼠妊娠期和哺乳期TFA暴露会导致子代肾脏损伤。膳食SDG干预可以延缓因母鼠妊娠期和哺乳期TFA暴露所导致的子代肾损伤，其作用机制可能通过作用于Bcl2/Bax/caspase-3信号轴通道，抑制细胞凋亡有关。

关键词: 亚麻木酚素, 反式脂肪酸, 慢性肾脏病, 营养干预, 细胞凋亡, Bcl2/Bax/caspase-3

Abstract:

Objective To investigate the potential mechanism underlying the protective effect of secoisolariciresinol diglucoside (SDG) against chronic kidney disease (CKD) in offspring mice caused by maternal exposure to trans fatty acids (TFA) during pregnancy and lactation. Methods Thirty female C57BL/6 mice were randomized into control group, TFA model group, and 3 TFA model groups treated with SDG at low, medium and high doses (10, 20 and 30 mg/kg, respectively). The changes in blood urea nitrogen (BUN) and serum creatinine (CRE) levels of the mice were measured. Network pharmacology analysis was conducted to explore protective mechanism of SDG against TFA-induced renal injury, and molecular docking was used to assess the binding affinity of SDG to Bcl-2, Bax, and caspase-3. The protein expressions of cleaved caspase-3, Bax, and Bcl-2 in the renal tissues of the offspring mice were detected with Western blotting. Result The mice in TFA group showed significantly higher BUN and CRE levels than those in the control group. Treatment with SDG at the medium and high doses significantly reduced BUN and CRE levels in the mouse models. Network pharmacology and molecular docking suggested that SDG ameliorated renal injury by targeting the apoptosis-related Bcl-2/Bax/caspase-3 axis. The results of Western blotting showed the mouse models in TFA exposure group had increased renal cell apoptosis with elevated expression levels of cleaved caspase-3 protein and a decreased Bcl-2/Bax ratio (P<0.05), and intervention with SDG at all the 3 doses significantly reduced renal cell apoptosis and renal expression of cleaved caspase-3 and increased the Bcl-2/Bax ratio in the mouse models. Conclusion Maternal TFA exposure during gestation and lactation induces renal injury in offspring mice. Dietary SDG intervention can mitigate TFA-induced renal injury in offspring mice possibly by suppressing renal cell apoptosis via regulating the Bcl-2/Bax/caspase-3 signaling axis.

Key words: secoisolariciresinol diglucoside, trans-fatty acid, chronic kidney disease, nutrition interventions, apoptosis, Bcl-2/Bax/caspase-3

马思雨, 陈美庆, 吴天宇, 赵文红. 膳食亚麻木酚素通过调控Bcl2/Bax/caspase3信号轴通道改善大鼠母体反式脂肪酸暴露所导致的子代肾损伤[J]. 南方医科大学学报, 2025, 45(12): 2658-2666.

Siyu MA, Meiqing CHEN, Tianyu WU, Wenhong ZHAO. Dietary secoisolariciresinol diglucoside alleviates chronic kidney disease in offspring rats caused by maternal trans-fatty acid exposure by regulating the Bcl-2/Bax/caspase-3 signaling axis[J]. Journal of Southern Medical University, 2025, 45(12): 2658-2666.

图/表 7

表1 各组子鼠血清肾功能指标比较

Tab.1 Comparison of blood urea nitrogen (BUN) and serum creatinine (CRE) levels among the 5 groups (Mean±SD, n=9)

Group	BUN (mmol/L)	CRE (μmol/L)
Control	17.08±1.53	44.85±6.087
TFA	22.39±6.18^#	68.54±11.89^#
TFA+LSDG	18.84±3.91	67.22±10.96
TFA+MSDG	15.56±1.86*	60.76±7.62
TFA+HSDG	17.01±3.57*	52.49±6.22
F	3.98	10.81
P	<0.01	<0.001

图1 SDG保护TFA诱导CKD的网络药理学分析

Fig.1 Network pharmacology analysis of the protective mechanism of SDG against TFA-induced chronic kidney disease (CKD). A: Collection of SDG targets. B: DO enrichment analysis of SDG targets. C: KEGG enrichment analysis of SDG targets. D: Screening of potential targets involved in the protective effects of SDG against TFA-induced CKD. E: KEGG enrichment analysis of the pathways mediating the protective effects of SDG against CKD.

图2 各组子鼠肾组织细胞凋亡情况

Fig.2 Cell apoptosis in the kidney tissue of the offspring mice in each group. A: TUNEL staining of the kidney tissue (the white arrows indicate apoptotic cells). B: Statistical chart of cell apoptosis in kidney tissue of the offspring mice in each group. #P<0.05 vs control group; *P<0.05 vs TFA group.

图3 各组子鼠肾组织Cleaved-caspase3的IHC染色情况

Fig.3 Immunohistochemical staining for cleaved caspase-3 in the renal tissues of the offspring mice in each group (scale bar=50 μm). A: Control group. B: TFA group. C: TFA+LSDG group. D: TFA+MSDG group. E: TFA+HSDG group. F: Quantitative statistical chart. #P<0.05 vs Control group; *P<0.05 vs TFA group.

图4 SDG治疗TFA诱导CKD的核心靶点筛选及分子对接结果

Fig.4 Screening of the key targets of SDG in the treatment of TFA-induced CKD (A) and molecular docking results of Bcl-2, Bax and caspase-3 with SDG (B-D).

表2 各蛋白质的PDB数据库代码名称

Tab. 2 The PDB database code names for each protein

Target	PDB ID
Bcl-2	1g5m
Bax	4bd6
Caspase-3	1gfw

图5 各组子鼠肾组织中细胞凋亡相关因子蛋白表达检测

Fig.5 Detection of protein expressions of apoptosis-related proteins in kidney tissue of the offspring mice in each group. A: Protein bands of caspase-3, Bcl-2 and Bax in the kidney tissues of the offspring mice in each group detected with Western blotting. B: Gray value of caspase-3 in each group. C: Gray value ratio of Bcl2/Bax in each group. #P<0.05 vs control group; *P<0.05 vs TFA group.

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