Protective effect of Tongdu Tiaoshen acupuncture combined with Xiaoxuming decoction (小续命汤) on dopaminergic neurons in Parkinson’s disease model

doi:10.19852/j.cnki.jtcm.20230214.005

Abstract

Abstract:

OBJECTIVE: To explore the possible mechanism of Tongdu Tiaoshen acupuncture combined with Xiaoxuming decoction (小续命汤, XXMD) in the treatment of Parkinson’s disease (PD).

METHODS: C57BL/6 mice were randomly divided into eight groups (n = 12), including blank group, model group, medication group, acupuncture group, high-dose XXMD group (XXMD-H), low-dose XXMD group (XXMD-L), acupuncture combined with high-dose XXMD group (A+H), and acupuncture combined with low-dose XXMD group (A+L). After treatment for 6 weeks, dopamine (DA) neurons and the pathological changes of tyrosine hydroxylase (TH) positive cells were observed. The enzyme-linked immunosorbent assay (ELISA) was used to measure the content of DA and the level of interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-10 (IL-10) and tumor necrosis factor alpha (TNF-α). The mRNA level of PINK1 and Parkin and the protein expression of Nix, PINK1 and Parkin in the substantia nigra were also detected.

RESULTS: Combination treatment effectively ameliorated the symptoms of PD. Compared with model group, combined treatment significantly up-regulated the protein expression of Nix, Parkin and PINK1 and the mRNA levels of PINK1 and Parkin in the substantia nigra (P<0.0001, P<0.001, P<0.01 or P<0.05). Furthermore, the levels of pro-inflammation cytokines were obviously decreased after combination therapy, while IL-10 content was increased remarkably (P<0.01).

CONCLUSION: Compared with each treatment alone, combination therapy improved the pathological damage of DA neurons of PD mice more effectively. The possible mechanism may be attributed to the up-regulated level of mitochondrial autophagy and improved mitochondrial function. These results provide fresh insight into the mechanism of co-treatment with Tongdu Tiaoshen acupuncture and XXMD for PD.

Key words: Parkinson disease, acupuncture, dopaminergic neurons, Tongdu Tiaoshen, Xiaoxuming decoction, mitochondrial autophagy

WU Haiyang, WANG Ying, HAN Wei, LI Huihui, JI Haisheng, LIU Xiuxiu. Protective effect of Tongdu Tiaoshen acupuncture combined with Xiaoxuming decoction (小续命汤) on dopaminergic neurons in Parkinson’s disease model[J]. Journal of Traditional Chinese Medicine, 2023, 43(3): 484-493.

Figures/Tables 4

Figure 1 Representative images of substantia nigra tissues and tyrosine hydroxylase (TH) in substantia nigra A1-H1: representative images of substantia nigra tissues (hematoxylin-eosin staining, ×200). A1: blank group; B1: model group; C1: XXMD-H group; D1: XXMD-L group; E1: acupuncture group; F1: A + H group; G1: A + L group; H1: medication group. Red arrows indicate severely damaged dopamine neurons. A2-H2: representative histologic images of TH in substantia nigra (immumohistochemical staining, ×400). A2: blank group; B2: model group; C2: XXMD-H group; D2: XXMD-L group; E2: acupuncture group; F2: A + H group; G2: A+L group; H2: medication group. The brown or brownish yellow in the picture is positive staining. Blank group: without any treatment. Model group: only modeled without any treatment. XXMD-H and XXMD-L group: treated with Xiaoxunming decotion by intragastric administration (6 weeks, 60 and 15 g·kg-1·d-1, respectively). A + H and A + L group: treated with Xiaoxunming decotion by intragastric administration (6 weeks, 60 and 15 g·kg-1·d-1, respectively) and corresponding acupuncture. Medication group: treated with dopasrazide (0.5 mg/10 g, 3 times/d, 6 weeks) by gavage. XXMD-H: high-dose Xiaoxunming decotion; XXMD-L: low-dose Xiaoxunming decotion; A + H: acupuncture combined with high-dose Xiaoxunming decotion; A + L: acupuncture combined with low-dose Xiaoxunming decotion.

Table 1 Locomotor counts and pole test scores of mice in each group

Group	n	Locomotor counts			Pole test scores
Group	n	Pre-modeling	Post-modeling	Post-treatment	Pre-modeling	Post-modeling	Post-treatment
Blank	12	148.143±17.332	135.600±15.231	143.773±14.121	0.073±0.0328	0.074±0.033	0.073±0.032
Model	12	148.791±18.508	81.237±9.830^a	84.782±11.725^a	0.057±0.041	1.924±0.208^a	1.936±0.311^a
XXMD-H	12	136.218±6.112	89.471±8.536^b	114.070±9.051^bc	0.072±0.032	1.751±0.192^a	0.592±0.109^bc
XXMD-L	12	139.212±9.537	78.207±10.738^a	99.718±8.741^a	0.059±0.034	1.729±0.136^a	0.711±0.086^bd
Acupuncture	12	132.031±10.229	91.288±10.126^b	118.187±10.513^c	0.064±0.043	1.691±0.110^a	0.721±0.127^ad
A+H	12	140.677±11.141	80.132±8.224^a	139.033±15.862^d	0.069±0.031	1.743±0.082^a	0.514±0.122^bd
A+L	12	135.571±14.329	92.379±8.010^b	120.881±15.722^c	0.058±0.027	1.677±0.142^a	0.583±0.120^ad
Medication	12	132.031±13.717	97.521±7.626^b	136.338±17.206^c	0.062±0.032	1.713±0.130^a	0.389±0.133^bd

Table 2 Comparison of serum levels of IL-1β, IL-6, IL-10 and TNF-α in each group (pg/mL)

Group	n	IL-1β	IL-6	IL-10	TNF-α
Blank	12	27±3	38±5	145±12	109±8
Model	12	106±12^a	126±11^a	40±5^a	339±26^a
XXMD-H	12	63±9^bc	73±7^bc	71±8^bc	243±29^bc
XXMD-L	12	73±8^bc	85±9^bd	64±9^bc	256±37^b
Acupuncture	12	67±8^bc	73±6^bc	67±5^bc	242±34^bc
A+H	12	41±7^d	54±5^de	104±12^cd	146±25^d
A+L	12	48±6^de	63±6^de	81±10^bc	212±20^bd
medication	12	39±5^d	52±7^d	107±8^cd	141±24^d

Figure 2 Expression of Nix, Parkin and PINK1 in the substantia nigra of mice after different treatment A: epresentative images of protein bands of Nix, Parkin and PINK1. B-D: results of quantitative analyses of Nix (B), Parkin (C) and PINK1 (D) in protein level. E, F: results of relative mRNA levels of PINK1 (E) and Parkin (F). Compared with model group, aP<0.0001, bP<0.001, cP<0.01, dP<0.05; compared with black group, aP<0.0001, bP<0.001, cP<0.01, dP<0.05. Blank group: without any treatment. Model group: only modeled without any treatment. XXMD-H and XXMD-L group: treated with Xiaoxunming decotion by intragastric administration (60 and 15 g·kg-1·d-1, respectively). A + H and A + L group: treated with Xiaoxunming decotion by intragastric administration (60 and 15 g·kg-1·d-1, respectively) and corresponding acupuncture. Medication group: treated with dopasrazide (0.5 mg/10 g) by gavage. XXMD-H: high-dose Xiaoxunming decotion; XXMD-L: low-dose Xiaoxunming decotion; A + H: acupuncture combined with high-dose Xiaoxunming decotion; A + L: acupuncture combined with low-dose Xiaoxunming decotion. 1: model; 2: blank 3: A + L; 4: A + H; 5: acupuncture; 6: XXMD-L; 7: XXMD-H; 8: medication. Statistical significance between the groups was determined by one-way analysis of variance. The sample size for each group was 12.

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