Beta-hydroxybutyric acid improves energy dysfunction of mouse hippocampal neuron HT22 cells induced by amyloid-β protein 1-42

doi:10.12307/2025.074

Abstract

Abstract: BACKGROUND: Patients with Alzheimer’s disease have severe brain energy disorders. In recent years, brain energy rescue strategies based on ketone body intervention have attracted more and more attention in the treatment of Alzheimer’s disease.
OBJECTIVE: To investigate whether β-hydroxybutyric acid can improve energy dysfunction by improving mitochondrial bioenergy function in HT22 cells of mouse hippocampal neurons induced by amyloid-β protein 1-42 (Aβ1-42).
METHODS: HT22 cells were divided into four groups: Control, β-hydroxybutyric acid, Aβ1-42, Aβ1-42+ β-hydroxybutyric acid. Related detection kits were respectively used to detect HT22 cell survival rate, adenosine triphosphate level, α-ketoglutarate dehydrogenase activity, Na+K+-ATPase activity, mitochondrial membrane potential, and reactive oxygen species levels.
RESULTS AND CONCLUSION: Compared with the control group, the survival rate, adenosine triphosphate level, α-ketoglutarate dehydrogenase activity, Na+K+-ATPase activity, and mitochondrial membrane potential of HT22 cells were significantly decreased (P < 0.05), and the level of reactive oxygen species was significantly increased (P < 0.05) in the Aβ1-42 group. Compared with the Aβ1-42 group, the survival rate, adenosine triphosphate level, α-ketoglutarate dehydrogenase activity, Na+K+-ATPase activity, and mitochondrial membrane potential of HT22 cells were significantly increased (P < 0.05), and the reactive oxygen species level was significantly decreased (P < 0.05) in the Aβ1-42+β-hydroxybutyric acid group. These results showed that β-hydroxybutyric acid improved mitochondrial bioenergetic function and ultimately improved Aβ1-42-induced energy impairment and survival rate in HT22 cells.

Key words: Alzheimer’s disease, HT22 cell, β-hydroxybutyric acid, mitochondrial bioenergetic function, energy dysfunction, adenosine triphosphate

CLC Number:

Ye Yucai, Fu Chaojing, Li Yan, Li Xinru, Chai Shifan, Cai Hongyan, Wang Zhaojun. Beta-hydroxybutyric acid improves energy dysfunction of mouse hippocampal neuron HT22 cells induced by amyloid-β protein 1-42[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(13): 2713-2719.

Figures/Tables 1

2.1 β-羟基丁酸改善Aβ1-42诱导的HT22细胞的存活率对照组、β-羟基丁酸组、Aβ1-42组、Aβ1-42+β-羟基丁酸组的细胞存活率分别为(101.080±1.738)%，(107.955±0.898)%，(42.375±1.916)%，(61.320±2.427)%。与对照组相比，β-羟基丁酸组的细胞存活率无统计学差异(P > 0.05)，Aβ1-42组的细胞存活率明显下降(P < 0.001)，提示Aβ1-42对HT22细胞具有损伤作用；与Aβ1-42组相比，Aβ1-42+β-羟基丁酸组的细胞存活率明显升高(P < 0.001)，提示β-羟基丁酸提高了Aβ1-42诱导的HT22细胞的存活率，见图1。 2.2 β-羟基丁酸增加Aβ1-42诱导HT22细胞的ATP水平，改善能量缺陷对照组、β-羟基丁酸组、Aβ1-42组、Aβ1-42+β-羟基丁酸组细胞的ATP相对浓度(nmol/mg)分别为24.644±1.179，43.547±1.423，13.369±0.069，42.653±2.294。与对照组相比，β-羟基丁酸组的ATP相对浓度明显升高(P < 0.001)，表明β-羟基丁酸促进ATP生成。与对照组相比，Aβ1-42组的ATP相对浓度明显下降(P < 0.01)；与Aβ1-42组相比，Aβ1-42+β-羟基丁酸组的ATP相对浓度明显升高(P < 0.001)，表明β-羟基丁酸增加了Aβ1-42诱导的HT22细胞的ATP水平，改善能量缺陷，见图2。 2.3 β-羟基丁酸增加Aβ1-42诱导HT22细胞的α-KGDH活性，促进ATP生成对照组、β-羟基丁酸组、Aβ1-42组、Aβ1-42+β-羟基丁酸组细胞的α-KGDH的相对活性(U/mg)分别为6.035±0.749，10.298±1.372，2.245±0.622，12.429±1.068。与对照组相比，β-羟基丁酸组的α-KGDH相对活性明显升高(P < 0.01)，表明β-羟基丁酸有利于ATP的生成。与对照组相比，Aβ1-42组的α-KGDH相对活性明显下降(P < 0.05)；与Aβ1-42组相比，Aβ1-42+β-羟基丁酸组的α-KGDH相对活性明显升高(P < 0.001)，提示β-羟基丁酸增加了Aβ1-42诱导的HT22细胞的α-KGDH活性，促进ATP生成，见图3。 2.4 β-羟基丁酸增加Aβ1-42诱导HT22细胞的Na+K+-ATP酶活性，促进ATP分解产能对照组、β-羟基丁酸组、Aβ1-42组、Aβ1-42+β-羟基丁酸组细胞Na+K+-ATP酶的相对活性(U/mg)分别为0.859±0.058，1.420±0.118，0.356±0.068，1.960±0.131。与对照组相比，β-羟基丁酸组的Na+K+-ATP酶相对活性升高(P < 0.05)，表明β-羟基丁酸有利于ATP分解产能。与对照组相比，Aβ1-42组Na+K+-ATP酶的相对活性下降(P < 0.05)；与Aβ1-42组相比，Aβ1-42+β-羟基丁酸组Na+K+-ATP酶的相对活性明显升高(P < 0.001)，提示β-羟基丁酸增加了Aβ1-42诱导的HT22细胞的Na+K+-ATP酶活性，促进ATP分解产能，有利于满足神经功能的高能量需求，见图4。 2.5 β-羟基丁酸增加Aβ1-42诱导HT22细胞的线粒体膜电位，改善线粒体生物能量功能在倒置荧光显微镜×100倍镜下观察线粒体膜电位，与对照组相比，β-羟基丁酸组的红色荧光明显增加，提示β-羟基丁酸促进线粒体膜电位升高。与对照组相比，Aβ1-42组的红色荧光明显减少，绿色荧光明显增加；与Aβ1-42组相比，Aβ1-42+β-羟基丁酸组的绿色荧光明显减少，红色荧光增加，提示Aβ1-42诱导线粒体膜电位受损，β-羟基丁酸提高了Aβ1-42诱导的HT22细胞的线粒体膜电位，见图5A。使用多功能酶标仪测量JC-1 Red/JC-1 Green值，对照组、β-羟基丁酸组、Aβ1-42组、Aβ1-42+β-羟基丁酸组细胞的JC-1 Red/JC-1 Green值分别为5.576±0.131，9.546±0.387，4.179±0.081，5.238±0.228。与对照组相比，β-羟基丁酸组的JC-1 Red/JC-1 Green值明显升高(P < 0.001)，表明β-羟基丁酸改善线粒体膜电位。与对照组相比，Aβ1-42组的JC-1 Red/JC-1 Green值明显下降(P < 0.01)；与Aβ1-42组相比，Aβ1-42+β-羟基丁酸组的JC-1 Red/JC-1 Green值明显升高(P < 0.05)，见图5B，提示β-羟基丁酸提高了Aβ1-42诱导的HT22细胞的线粒体膜电位，改善线粒体生物能量功能。 2.6 β-羟基丁酸降低Aβ1-42诱导HT22细胞的活性氧水平，改善氧化应激在倒置荧光显微镜的×100倍镜下观察活性氧水平，与对照组相比，Aβ1-42组的绿色荧光明显增加；与Aβ1-42组相比，Aβ1-42+β-羟基丁酸组的绿色荧光明显减少，提示Aβ1-42诱导活性氧增加，β-羟基丁酸降低了Aβ1-42诱导的HT22细胞的活性氧水平，见图6A。使用多功能酶标仪测量细胞活性氧荧光值，对照组、β-羟基丁酸组、Aβ1-42组、Aβ1-42+β-羟基丁酸组细胞的活性氧相对荧光值(AU)分别为0.623±0.071，0.572±0.108，2.208±0.098，0.992±0.088。与对照组相比，β-羟基丁酸组的活性氧水平无统计学差异(P > 0.05)，Aβ1-42组的活性氧水平明显增加(P < 0.001)，提示Aβ1-42对HT22细胞具有氧化损伤作用；与Aβ1-42组相比，Aβ1-42+β-羟基丁酸组的活性氧水平明显降低(P < 0.001)，见图6B，提示β-羟基丁酸降低Aβ1-42诱导的HT22细胞的活性氧水平，有利于改善氧化应激。"

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