Mechanism of nodakenin in relieving neuropathic pain

doi:10.11958/20240971

Abstract

Abstract:

Objective To investigate the effect and mechanism of nodakenin (Nod) in neuropathic pain (NP).Methods Differential expression genes in the primary somatsensory cortex (S1) of NP data and overlapping genes between the dataset and mitochondrial data were screened and analyzed. Overlapping gene interaction networks were overlapped and core genes were screened. A total of 27 mice were randomly divided into the sham operation group, the model group and the drug administration group (9 mice/group). The chronic compression injury model of sciatic nerve was constructed in the model group and the drug administration group. Nod 10 mg/kg was intraperitoneally injected into the drug administration group for 1 week. Changes of pain behavior and motor ability in mice were detected. HE staining and Nissl staining were used to detect effects of nerve injury and inflammation on brain tissue of S1 region of mice. The expression levels of interleukin-1β, early gene (c-Fos), panthenol-cytochrome c reductase complex III subunit (Uqcrq) and ubiquinone oxidoreductase subunit (Nduf) b5 in S1 brain region were analyzed by Western blot assay. Molecular docking was used to study the target of Nod. PC12 cells were divided into the control group, the IL-1β group (1 μmol/L IL-1β treatment) and the IL-1β+Nod group (1 μmol/L IL-1β+1 μmol/L Nod treatment), and mitochondrial membrane potential was detected in each group.Results In the NP dataset GSE180627, S1 brain region contained 293 differentially expressed genes, and the mitochondrial data contained 1 082 genes. There were 34 overlapping genes, and genes related to oxidative phosphorylation and electron transport chain were enriched. The protein interaction network showed that core genes included electron transport chain related proteins Ndufb5, Uqcrq, Ndufs8, Ndufa7, Ndufa3, Cox6b1 and Mrps33. Compared with the model group, the mechanical foot shrinkage threshold, thermal foot shrinkage reflex latency and rod rotation residence time of mice were increased in the drug administration group, the number of inflammatory infiltrating cells in S1 tissue and the number of Nislet bodies in neurons, expression levels of c-Fos and IL-1β in neurons were decreased, and expression levels of Uqcrq and Ndufb5 were increased (P<0.05). Molecular docking showed that Nod could bind Uqcrq and Ndufb5. Compared with the IL-1β group, the fluorescence signal of mitochondrial membrane potential was enhanced in the IL-1β+Nod group (P<0.05).Conclusion Nodakenin can improve pain behavior in mice, and its mechanism involves ameliorating mitochondrial damage in S1.

Key words: Nodakenin, mitochondria, inflammation, neuropathic pain, primary somatosensory cortex

CLC Number:

R965

Figures/Tables 10

Fig.1 Analysis of NP and mitochondrial differential expression genes

Tab.1 Comparison of behavioral tests between three groups of mice （n=9，$\bar{x}$±s）

组别	PWT/g	TWL/s	转棒停留时间/s
假手术组	1.00±0.32	15.40±3.01	526.67±62.11
模型组	0.33±1.77^a	5.89±1.95^a	129.14±48.84^a
给药组	0.57±0.17^ab	10.89±1.69^ab	277.73±81.23^ab
F	19.550^＊＊	38.790^＊＊	84.826^＊＊

Fig.2 HE staining of S1 cortex of mice in each group

Fig.3 Nissl staining of S1 cortex of mice in each group

Tab.2

组别	炎症细胞	尼氏小体
假手术组	6.66±0.81	8.83±1.83
模型组	14.16±1.94^a	16.00±1.78^a
给药组	9.00±0.89^ab	10.50±1.37^b
F	50.670^＊＊	29.902^＊＊

Fig.4 Protein expression of proinflammation cytokines IL-1β and c-Fos in each group

Tab.3 Comparison of proinflammation cytokines IL-1β and c-Fos protein expression between the three groups （n=3，$\bar{x}$±s）

组别	c-Fos	IL-1β
假手术组	1.00±0.08	1.00±0.06
模型组	1.32±0.07^a	2.07±0.06^a
给药组	1.09±0.07^ab	1.81±0.05^ab
F	13.113^＊＊	195.269^＊＊

Fig.5 Schematic diagram of molecular docking results and Western blot assay of Uqcrq and Ndufb5 protein in each group

Tab.4 Comparison of proinflammation cytokines Ndufb5 and Uqcrq protein expression between the three groups （n=3，$\bar{x}±s$）

组别	Ndufb5	Uqcrq
假手术组	1.00±0.05	1.00±0.04
模型组	0.68±0.05^a	0.89±0.05^a
给药组	1.14±0.05^ab	1.08±0.04^b
F	73.705^＊＊	12.909^＊＊

Fig.6 Fluorescence intensity of each group of cells (×400)

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