Pharmacodynamics of Qingxin Jieyu Granules for treatment of atherosclerosis and its regulatory mechanism for lipid metabolism

doi:10.12122/j.issn.1673-4254.2024.08.10

Abstract

Abstract:

Objective To elucidate the therapeutic mechanism of Qingxin Jieyu Granule (QXJYG) against atherosclerosis (AS) based on network pharmacology. Methods The major targets and pathways of QXJYG against AS were analyzed using network pharmacology. Rat models of AS established by high-fat feeding combined with intraperitoneal vitamin D3 injection were treated daily with normal saline, atorvastatin (13.15 mg/kg), or QXJYG at 0.99, 1.98, and 3.96 g/kg for 8 weeks (n=6). Ultrasound and HE staining were used to assess the function and pathologies of the abdominal aorta. Blood lipids and serum levels of Ang II, ET-1, TXA2, PGI2, and ox-LDL of the rats were detected using an automatic biochemical analyzer or ELISA. The expressions of LOX-1, PPARγ, RXRα, p-P65, VCAM-1 and ICAM-1 in the abdominal aorta were detected with immunohistochemistry. Results The rat models of AS showed obvious abdominal aorta wall thickening, increased pulse wave velocity and pulse index, decreased inner diameter of the abdominal aorta, elevated levels of TC, LDL-C, Ang II, ET-1 and TXA2, and lowered levels of HDL-C and PGI2. QXJYG and atorvastatin treatment of the rat models significantly alleviated histopathological changes of the abdominal aorta, decreased serum levels of TC, LDL-C, Ang II, ET-1 and TXA2, and increased the levels of HDL-C and PGI2. Network pharmacology study suggested the therapeutic effect of QXJYG against AS was mediated by regulating lipid metabolism, PPAR and NF‑κB pathways. Consistently, treatments with QXJYG were found to significantly decrease ox-LDL level and LOX-1, P-P65, VCAM-1 and ICAM-1 protein expressions while increasing PPARγ and RXRα expressions in the aorta of AS rats. Conclusion QXJYG alleviates lipid metabolism disorder and improves histopathological changes of the abdominal aorta of AS rats possibly by lowering ox-LDL level, reducing LOX-1 expression, activating PPARγ and RXRα, and inhibiting P65 phosphorylation to reduce VCAM-1 and ICAM-1 expression in the aorta.

Key words: Qingxin Jieyu Granule, atherosclerosis, network pharmacology, PPARγ/RXRα signaling pathway, NF-κB signaling pathway

Shanyuan ZHANG, Qiaoyan CAI, Jianghan QI, Kaixin YIN, Chenchen HE, Zhuye GAO, Ling ZHANG, Jianfeng CHU. Pharmacodynamics of Qingxin Jieyu Granules for treatment of atherosclerosis and its regulatory mechanism for lipid metabolism[J]. Journal of Southern Medical University, 2024, 44(8): 1518-1528.

Figures/Tables 10

Fig.1 Effects of QXJYG on vessel wall thickness, pulse wave propagation velocity, pulsatility index and vessel diameter in atherosclerotic rats. A, B: Ultrasound images and quantitative comparison of wall thickness of the abdominal aorta. C, D: Sonograms and quantitative comparison of pulse-wave propagation velocity of the abdominal aorta. E, F: Echocardiograms and quantitative analysis of the pulsatility index of the abdominal aorta. G, H: Ultrasound images and quantitative analysis of abdominal aorta vessel diameter. LPT: Positive control (atorvastatin) group; LD: Low-dose QXJYG group; MD: Medium-dose QXJYG group; HD: High-dose QXJYG group. *P<0.05 vs Control group; #P<0.05 vs Model group; △P<0.05 vs LD group; &P<0.05 vs MD group.

Fig.2 Effects of QXJYG on vascular pathology of the abdominal aorta in rats with atherosclerosis (HE staining, Scale bar=50 µm).①Intimal thickening of the blood vessels; ②Proliferation of smooth muscle cells with disordered arrangement.

Fig.3 Effects of QXJYG on serum levels of TC, LDL-C, TG and HDL-C in rats with atherosclerosis. A: Serum levels of TC and LDL-C. B: Serum levels of TG and HDL-C. *P<0.05 vs Control group; #P<0.05 vs Model group; △P<0.05 vs LD group.

Fig.4 Effects of QXJYG on serum levels of Ang II (A), ET-1 (B), TXA2 (C) and PGI2 (D) in rats with atherosclerosis. *P<0.05 vs Control group; #P<0.05 vs Model group; △P<0.05 vs LD group; &P<0.05 vs MD group.

Fig.5 Analysis of targets of QXJYG and AS. A: Quantity diagram of each drug compound of QXJYG. B: Number of drug targets of QXJYG. C: Venn diagram of QXJYG and AS targets. D: PPI plot of potential protein interaction of QXJYG against AS.

Tab.1 Cross targets of QXJYG with anti-AS values higher than the average value

Key target	Degree	Betweenness centrality	Closeness centrality
IL-6	21	0.14771628	0.83870968
IL-1B	19	0.0880959	0.78787879
PPARG	19	0.20820912	0.78787879
VCAM1	16	0.03766667	0.7027027
IL-10	14	0.02131169	0.66666667
SERPINE1	14	0.01293806	0.65
ESR1	14	0.04946542	0.65
SPP1	14	0.01452414	0.65
CRP	13	0.00772194	0.63414634
SIRT1	13	0.04028405	0.65
NOS3	13	0.01529604	0.63414634
MMP3	11	0.00423576	0.57777778
SELE	10	0.00117216	0.59090909

Fig.6 KEGG pathway enrichment analysis and GO functional analysis of QXJYG against AS.A: KEGG pathway enrichment analysis of QXJYG against AS. B: Biological processes analysis of QXJYG against AS. C: Cell components analysis of QXJYG against AS. D: Molecular function analysis of QXJYG against AS.

Fig.7 Effect of QXJYG on serum ox-LDL in atherosclerotic rats. *P<0.05 vs Control group; #P<0.05 vs Model group; △P<0.05 vs LD group.

Fig.8 Effect of QXJYG on the expression of LOX-1, PPARγ, RXRα and p-P65 in the abdominal aorta of atherosclerotic rats (DAB staining, original magnification: ×400). A, B: LOX-1 expression and positivity rate in the abdominal aorta. C, D: PPARγ expression and positivity rate in the abdominal aorta. E, F: Expression of RXRα and positivity rate positive rate. G, H: Expression of p-P65 and positivity rate. *P<0.05 vs Control group; #P<0.05 vs Model group; △P<0.05 vs LD group; &P<0.05 vs MD group.

Fig.9 Effect of QXJYG on the expression of VCAM-1 and ICAM-1 in abdominal aorta of atherosclerotic rats (DAB staining, ×400). A, B: VCAM-1 expression and positivity rate in the abdominal aorta. C, D: ICAM-1 expression and positivity rate in the abdominal aorta. *P<0.05 vs Control group; #P<0.05 vs Model group; △P<0.05 vs LD group.

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