Congrong Shujing Granules Ameliorates Mitochondrial Associated Membranes to Against MPP + -Induced Neurological Damage in the Cellular Model of Parkinson's Disease

Chutian Zhang^1,2*Peizhen Huang¹Huiling Cheng¹Xinxin Yuan¹Mei Zhou¹Ying Liu¹Ting Liu³Dan Chen¹Qian Xu¹Jing Cai^1,3*

• ¹Fujian University of Traditional Chinese Medicine, Fuzhou, China
• ²Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan, Hubei Province, China
• ³Third Affiliated People's Hospital of Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian Province, China

Context: Congrong-Shujing Granules (CRSJG) are known to protect dopaminergic neurons in Parkinson's disease (PD), and the mechanism may be related to the improvement of mitochondria-associated membranes (MAMs). Objective: To investigate the impact of CRSJG-medicated serum on MAMs in the 1-methyl-4-phenylpyridinium (MPP⁺)-induced PD cell model. Materials and methods: Human neuroblastoma (SH-SY5Y) cells were treated with 1000 μmol/L MPP⁺ for 24 h, resulting in three experimental groups: MPP⁺, CRSJG, and 2-APB. The MPP⁺ group received blank serum, CRSJG group was treated with CRSJG-medicated serum, and the 2-APB group was given 100 μmol/L 2-APB. An untreated control group was also included. Serum pharmacokinetics for the CRSJG group were analyzed using ultra-performance liquid chromatography-tandem mass spectrometry. Intracellular-Ca²⁺, reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and apoptosis rates were measured using fluorescent probes and flow cytometry. Transmission electron microscopy was employed to examine the morphology of MAMs. Western blot was conducted to identify proteins related to apoptosis and Ca²⁺ transport. Results: CRSJG-medicated serum identified by pharmacokinetic markers including echinacoside, paeoniflorin, salvianolic acid B, acteoside, and tanshinone IIa. The EC50 for MPP + -induced reduction in cell proliferation was 1110 μmol/L. CRSJG-medicated serum, especially at 2.5%, significantly improved cell proliferation after 24 h. The serum effectively mitigated damage within the MAMs region and reduced both the mitochondrial-Ca²⁺ fluorescence intensity and the expression of the IP₃R-VDAC-MCU complex in MPP⁺-induced neuronal cells. Additionally, it significantly decreased the levels of ROS, the decline in MMP, and apoptosis rates in these cells. Discussion and Conclusion: The findings provide novel insights into the potential of CRSJG in treating neuronal loss in PD.