Tianma Gouteng Granule improves motor deficits in mouse models of Parkinson's disease by regulating the necroptosis pathway

doi:10.12122/j.issn.1673-4254.2025.08.01

Abstract

Abstract:

Objective To investigate the effects of formulated granules of Tianma Gouteng Yin (TGY) on motor deficits in a mouse model of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced subacute Parkinson's disease (PD) and explore the possible molecular mechanisms. Methods Ninety C57BL/6 mice were randomized equally into 6 groups, including a control group, a PD model group, a NEC-1 (6.5 mg/kg) treatment group, two TGY treatment groups at 5 and 2.5 g/kg, and a Madopar (76 mg/kg) treatment (positive control) group. Mouse models of PD were established by intraperitoneal injection of MPTP (30 mg/kg) for 5 consecutive days with the corresponding treatments for 15 days. The mice were randomly selected for motor function tests. Western blotting was used to detect the changes in expressions of TH, α-syn, RIPK1, RIPK3 and MLKL in the striatum of the mice. Network pharmacology analysis and molecular docking studies were performed to explore TGY-mediated regulation of the necroptosis pathway for PD treatment. Results Compared with those in the control group, the PD model mice exhibited obvious motor deficits with significantly increased α-syn protein expression and lowered TH protein expression in the striatum. Treatment with NEC-1 obviously improved motor deficits, inhibited the necroptosis pathway, and alleviated the changes in TH and α‑syn proteins in PD mice. Network pharmacology and molecular docking analyses suggested that the therapeutic effect of TGY in PD was associated with the modulation of RIPK1, a key protein in the necroptosis pathway. In PD mouse models, TGY treatment at the two doses significantly improved motor deficits of the mice, increased TH expression, and decreased the expressions of α-syn and necroptosis-related proteins in the striatum. Conclusion TGY can effectively inhibit the necroptosis pathway, increase TH expression and decrease α-syn expression in the striatum to improve motor deficits in PD mice.

Key words: Parkinson's disease, Tianma Gouteng Yin, motor deficits, necroptosis pathway

Dandan CHEN, Qianqian REN, Menglin LÜ, Baowen ZHANG, Xingran LIU, Meng ZHANG, Yang WANG, Xianjuan KOU. Tianma Gouteng Granule improves motor deficits in mouse models of Parkinson's disease by regulating the necroptosis pathway[J]. Journal of Southern Medical University, 2025, 45(8): 1571-1580.

Figures/Tables 5

Fig. 1 Establishment of mouse models of MPTP/p-induced Parkinson's disease (PD). A: Schematic diagram of mice tested on the runway. B: Gait pattern and stride length in the control group. C: Gait pattern and stride length in the PD model group. D: Quantification of step cycle. E: Quantification of stride length. F: Expression of TH protein and a-syn protein in the striatum of the mice. Data are presented as Mean±SD if not specified otherwise and analyzed using one-way ANOVA. ##P<0.01,###P<0.001 vs Control. LF: Left front paw. RF: Right front paw. LH: Left hind. RH: Right hind.

Fig.2 Tianma Gouteng Yin (TGY) at low (TGY-L) and high doses (TGY-H) improves motor deficits in PD mice by. A: Quantification of step cycle. B: Quantification of stride length. C: Quantification of latency to fall in rotarod test. D: Quantification of T-total in pole test. E: Quantification of T-turn in pole test. *P<0.05, **P<0.01, ***P<0.001 vs PD; ^P<0.01, ^^P<0.01, ^^^P<0.01 vs PD+TDY-L.

Fig.3 Prediction results of network pharmacology. A: TGY and necroptosis intersection genes. B: PPI network diagram of the core target proteins. C: Bubble map of KEGG analysis of the key genes. D: Molecular docking result patterns of RIPK1 and compounds in TGY.

Fig.4 NEC-1 attenuates motor deficits and pathological protein accumulation in PD mice. A: Quantification of latency to fall in rotarod test. B: Quantification of T-total in pole test. C: Quantification of T-turn in pole test. D: Expression of key proteins of the necroptosis pathway in the striatum of the mice. E: Expression of TH and a-syn protein in the striatum of the mice. #P<0.05, ###P<0.001 vs Control; *P<0.05, **P<0.01, ***P<0.001 vs PD.

Fig.5 TGY attenuates the necroptosis pathway and pathological protein accumulation in PD mice. A: Expression of the key proteins of the necroptosis pathway in the striatum of the mice. B: Expression of TH protein and a-syn protein in the striatum of the mice.*P<0.05, **P<0.01, ***P<0.001 vs PD.

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