Neuroprotective effects of Longxue Tongluo Capsule on ischemic stroke rats revealed by LC-MS/MS-based metabolomics approach

Jing Sun; Xianyang Chen; Yongru Wang; Yuelin Song; Bo Pan; Bei Fan; Fengzhong Wang; Xiaonan Chen; Pengfei Tu; Jiarui Han; Huixia Huo; Jun Li

doi:10.1016/j.chmed.2022.12.010

Neuroprotective effects of Longxue Tongluo Capsule on ischemic stroke rats revealed by LC-MS/MS-based metabolomics approach

Chin Herb Med. 2023 May 25;15(3):430-438. doi: 10.1016/j.chmed.2022.12.010. eCollection 2023 Jul.

Authors

Jing Sun^{1

2}, Xianyang Chen³, Yongru Wang², Yuelin Song¹, Bo Pan¹, Bei Fan², Fengzhong Wang², Xiaonan Chen¹, Pengfei Tu¹, Jiarui Han³, Huixia Huo¹, Jun Li¹

Affiliations

¹ Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China.
² Key Laboratory of Agro-products Quality and Safety Control in Storage and Transport Process, Laboratory of Agro-products Quality Safety Risk Assessment, Ministry of Agriculture, Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
³ Baofeng Key Laboratory of Genetics and Metabolism, Zhongguancun Biological and Medical Big Data Center, Beijing 101300, China.

Abstract

Objective: The present study aimed to evaluate the therapeutic effect and explore the underlying mechanisms of Longxue Tongluo Capsule (LTC) on ischemic stroke rats.

Methods: Twenty-six rats were randomly divided into four groups, including sham group, sham + LTC group, MCAO group, and MCAO + LTC group. Ischemic stroke rats were simulated by middle cerebral artery occlusion (MCAO), and LTC treatment group were orally administrated with 300 mg/kg of LTC once daily for seven consecutive days. LTC therapy was validated in terms of neurobehavioral abnormality evaluation, cerebral infarct area, and histological assessments. The plasma metabolome comparisons amongst different groups were conducted by UHPLC-Q Exactive MS in combination with subsequent multivariate statistical analysis, aiming to finding the molecules in respond to the surgery or LTC treatment.

Results: Intragastric administration of LTC significantly decreased not only the neurobehavioral abnormality scores but also the cerebral infarct area of MCAO rats. The interstitial edema, atrophy, and pyknosis of glial and neuronal cells occurred in the infarcted area, core area, and marginal area of cerebral cortex were improved after LTC treatment. A total of 13 potential biomarkers were observed, and Youden index of 11 biomarkers such as LysoPC, SM, and PE were more than 0.7, which were involved in neuroprotective process. The correlation and pathway analysis showed that LTC was beneficial to ischemic stroke rats via regulating glycerophospholipid and sphingolipid metabolism, together with nicotinate and nicotinamide metabolism. Heatmap and ternary analysis indicated the synergistic effect of carbohydrates and lipids may be induced by flavonoid intake from LTC.

Conclusion: The present study could provide evidence that metabolomics, as systematic approach, revealed its capacity to evaluate the holistic efficacy of TCM, and investigate the molecular mechanism underlying the clinical treatment of LTC on ischemic stroke.

Keywords: Longxue Tongluo Capsule; UHPLC-Q Exactive MS; ischemic stroke; metabolomics; middle cerebral artery occlusion; phenolic compounds.