Efficacy of Jiangzhi Xiaoban tablet (降脂消斑片) on toll-like receptor 4/nuclear factor-kappa B/nod-like receptor protein 3 signaling pathway in mice with atherosclerosis induced by high-fat diet

doi:10.19852/j.cnki.jtcm.20231121.002

Journal of Traditional Chinese Medicine ›› 2024, Vol. 44 ›› Issue (1): 88-94.DOI: 10.19852/j.cnki.jtcm.20231121.002

• Original articles • Previous Articles Next Articles

Efficacy of Jiangzhi Xiaoban tablet (降脂消斑片) on toll-like receptor 4/nuclear factor-kappa B/nod-like receptor protein 3 signaling pathway in mice with atherosclerosis induced by high-fat diet

LIU Huihui¹, FENG Jun²(), LIU Jianhe³, CHENG Choufu³, HU Guoheng⁴

1 Hunan University of Chinese Medicine, Changsha 410208, China
2 Department of Geriatrics, the First Hospital of Hunan University of Chinese Medicine, Changsha 410007, China
3 Department of Cardiology, the First Hospital of Hunan University of Chinese Medicine, Changsha 410007, China
4 Department of Neurology, the First Hospital of Hunan University of Chinese Medicine, Changsha 410007, China

Received:2022-11-02 Accepted:2023-02-02 Online:2024-02-15 Published:2023-11-21
Contact: FENG Jun, Department of Geriatrics, the First Hospital of Hunan University of Chinese Medicine, Changsha 410007, China. Naohvc@139.com. Telephone: +86-15211146864
Supported by:
Youth Fund Project of Hunan Natural Science Foundation Committee: to Explore the Protective Effect of Chaihu Sanshen Capsule on Myocardial Ischemia Reperfusion Injury Based on Iron Death and Inflammatory Response of Myocardial Cells Induced by Mixed lineage kinase 3 Signaling Pathway(2021JJ40419);Traditional Chinese Medicine Administration of Hunan Province: to Explore the Mechanism of the Protective Effect of Chaihu Sanshen Capsule on Myocardial Ischemia Reperfusion Injury based on MicroRNA-145-5p Inhibition of Iron Death and Inflammatory Response of Myocardial Cells Induced by Mixed Lineage Kinase 3 Signaling Pathway(B2023019);Health Commission of Hunan Province: to Explore the Mechanism of Chaihu Sanshen Capsule in Reducing Myocardial Ischemia Reperfusion Injury Based on Nuclear Factor Erythroid 2-related Factor 2/Heme Oxygenase-1 Pathway Inhibiting Iron Death of Myocardial Cells(2021116001377);Hunan University of Chinese Medicine: To Explore the Intervention Effect of Zhenwu Decoction on the Model of Heart and Kidney Yang Deficiency in Rats with Cardiorenal Syndrome through MicroRNA-214/Receptor-interacting Protein Kinase 1-mediated Tumor Necrosis Factor Signal Pathway(2020XJJJ038)

Abstract

Abstract:

OBJECTIVE: To study the effect of Jiangzhi Xiaoban tablet (降脂消斑片, JZXB) on toll-like receptor 4 (TLR4)/nuclear factor-kappa B (NF-κB)/Nod-like receptor protein 3 (NLRP3) signaling pathway expression in atherosclerosis (AS) mice by establishing a mouse model of AS, and to explore its mechanism of prevention and treatment of AS.

METHODS: Sixty-four male C57BL/6J mice were randomly divided into two groups, 12 in the normal control group and 52 in the model group (MOD). Seven weeks later, two mice in each of the above two groups were randomly sacrificed, and the whole aortic tissue of the mice was taken out for hematoxylin-eosin staining. After successful modeling, 50 mice in the modeling group were randomly divided into 5 groups: MOD, atorvastatin group (ATO), low-dose group of JZXB (JZXB-L), middle-dose group of JZXB (JZXB-M), and high-dose group of JZXB (JZXB-H), 10 mice in each group. The mice in each group were killed after 6 weeks of preventive administration. HE staining was used to observe the pathological changes of aorta in AS mice. The levels of serum triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were detected by automatic biochemical analyzer. The levels of inflammatory factor interleukin-1β (IL-1β) were detected by enzyme linked immunosorbent assay. The expression of TLR4, NF-κB and NLRP3 proteins in aortic tissue was detected by immunohistochemistry.

RESULTS: Compared with the MOD, the levels of serum TC, TG and LDL-C in the JZXB-H and ATO were significantly decreased, while the level of HDL-C was significantly increased. The levels of serum TG, LDL-C in the JZXB-M were significantly decreased, and the level of HDL-C was significantly increased. Compared with the MOD, the levels of IL-1β were significantly decreased, aortic lesions were significantly improved, and the expression of TLR4, NF-κB, and NLRP3 proteins in the aortic tissue was significantly decreased in the JZXB-H, JZXB-M, and ATO.

CONCLUSION: JZXB has inhibitory effect on atherosclerosis in mice, and its mechanism may be through regulating the TLR4/NF-κB/NLRP3 signaling pathway and reducing the inflammatory response, so as to play a role in inhibiting atherosclerosis.

Key words: atherosclerosis, toll-like receptor 4, NF-kappa B, NLR proteins, signal transduction, inflammation, Jiangzhi Xiaoban tablet

LIU Huihui, FENG Jun, LIU Jianhe, CHENG Choufu, HU Guoheng. Efficacy of Jiangzhi Xiaoban tablet (降脂消斑片) on toll-like receptor 4/nuclear factor-kappa B/nod-like receptor protein 3 signaling pathway in mice with atherosclerosis induced by high-fat diet[J]. Journal of Traditional Chinese Medicine, 2024, 44(1): 88-94.

Figures/Tables 6

Figure 1 Modeling of AS mice in each group (HE staining, ×100), scale bar 50 μm A: selected from the CON group fed with ordinary diet for 7 weeks; B: selected from the MOD group fed with high-fat diet for 7 weeks. CON: normal control; MOD: model; AS: atherosclerosis; HE: hematoxylin-eosin.

Figure 2 Effect of JZXB on the histopathology of aorta in AS mice (HE staining, × 400), scale bar 50 μm A, B: selected from the CON (A) and MOD group (B) received 0.5% sodium carboxymethyl cellulose aqueous solution at1.5 mL·kg-1·d-1 by gavage for 6 weeks; C: selected from the ATO group received atorvastatin at 4.1 mg·kg-1·d-1 by gavage for 6 weeks; D: selected from the JZXB-L group received JZXB at 492 mg·kg-1·d-1 by gavage for 6 weeks; E: selected from the JZXB-M group received JZXB at 984 mg·kg-1·d-1 by gavage for 6 weeks; F: selected from the JZXB-H group received JZXB at 1968 mg·kg-1·d-1 by gavage for 6 weeks. CON: normal control; MOD: model; ATO: atorvastatin; JZXB-L: low-dose of JZXB; JZXB-M: middle-dose of JZXB; JZXB-H: high-dose of JZXB; JZXB: Jiangzhi Xiaoban tablet; AS: atherosclerosis; HE: hematoxylin-eosin.

Table 1 Effect of Jiangzhi Xiaoban tablet on serum TC, TG, HDL-C and LDL-C levels in AS mice (mmol/L, $\bar{x}±s$)

Group	n	TC	TG	HDL-C	LDL-C
CON	7	2.24±0.15	0.99±0.17	2.09±0.24	0.45±0.1
MOD	7	3.49±0.31^a	1.70±0.27^a	1.41±0.18^a	1.81±0.61^a
ATO	7	2.29±0.24^b	0.98±0.16^b	2.19±0.28^b	0.76±0.13^b
JZXB-L	7	3.89±0.35	0.94±0.23^b	1.45±0.21	1.15±0.24^b
JZXB-M	7	4.03±0.43	0.98±0.18^b	2.31±0.19^b	0.82±0.16^b
JZXB-H	7	2.72±0.37^b	0.94±0.23^b	2.42±0.31^b	0.84±0.22^b

Table 2 Effect of JZXB on the level of IL-1β in AS mice ($\bar{x}±s$)

Group	n	IL-1β (pg/mL)
CON	7	22.1±0.9
MOD	7	34.4±2.9^a
ATO	7	25.7±2.1^b
JZXB-L	7	32.8±3.4
JZXB-M	7	26.1±2.2^b
JZXB-H	7	26.2±2.2^b

Table 3 Average optical density values of TLR4, NF-κB and NLRP3 protein expression in immunohistochemistry of mice in each group ($\bar{x}±s$)

Group	n	TLR4	NF-κB	NLRP3
CON	5	0.193±0.005	0.197±0.005	0.195±0.005
MOD	5	0.229±0.006^a	0.265±0.003^a	0.243±0.004^a
ATO	5	0.208±0.007^b	0.225±0.006^b	0.217±0.006^b
JZXB-L	5	0.228±0.005	0.260±0.004	0.238±0.007
JZXB-M	5	0.210±0.007^b	0.222±0.006^b	0.216±0.004^b
JZXB-H	5	0.213±0.006^b	0.227±0.005^b	0.221±0.006^b

Figure 3 Effect of JZXB on TLR4, NF-κB and NLRP3 protein expression levels in aortic tissue of AS mice (n = 5, × 400) A1-A6: protein expression level of TLR4 by immunohistochemistry; B1-B6: protein expression level of NF-κB by immunohistochemistry; C1-C6: protein expression level of NLRP3 by immunohistochemistry; A1, A2, B1, B2, C1, C2: selected from the CON (A1, B1, C1) and MOD group (A2, B2, C2) received 0.5% sodium carboxymethyl cellulose aqueous solution at 1.5 mL·kg-1·d-1 by gavage for 6 weeks; A3, B3, C3: selected from the ATO group received atorvastatin at 4.1 mg·kg-1·d-1 by gavage for 6 weeks; A4, B4, C4: selected from the JZXB-L group received JZXB at 492 mg·kg-1·d-1 by gavage for 6 weeks; A5, B5, C5: selected from the JZXB-M group received JZXB at 984 mg·kg-1·d-1 by gavage for 6 weeks; A6, B6, C6: selected from the JZXB-H group received JZXB at 1968 mg·kg-1·d-1 by gavage for 6 weeks. CON: normal control; MOD: model; ATO: atorvastatin; JZXB-L: low-dose of JZXB; JZXB-M: middle-dose of JZXB; JZXB-H: high-dose of JZXB; JZXB: Jiangzhi Xiaoban tablet; AS: atherosclerosis; TLR4: toll-like receptor 4; NF-κB: nuclear factor-kappa B; NLRP3: nod-like receptor protein 3.

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Efficacy of Jiangzhi Xiaoban tablet (降脂消斑片) on toll-like receptor 4/nuclear factor-kappa B/nod-like receptor protein 3 signaling pathway in mice with atherosclerosis induced by high-fat diet

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