Jianpi Qutan Fang (健脾祛痰方) induces anti-atherosclerosis and ameliorates endothelial cell injury in high-fat diet rats via an anti-inflammatory and inhibiting Janus kinase/signal transducer and activator of transcription signaling pathway

doi:10.19852/j.cnki.jtcm.20230814.002

Abstract

Abstract:

OBJECTIVE: To investiage the possible mechanism underlying the effect of the Jianpi Qutan Fang (健脾祛痰方, JPQT) on Atherosclerosis (AS) which is the main pathological process of most cardiovascular diseases that affect millions of adults worldwide.
METHODS: In the present study, rats were fed with a high-fat-diet (HFD) with vitamin D3 for 16 weeks and were orally administered atorvastatin treatment and different doses of JPQT. Histopathological changes and ultrastructural changes in the aorta were evaluated through hematoxylin-eosin staining and transmission electron microscopy (TEM), respectively. Suppressor of cytokine signaling 1 (SOCS1)/Janus kinase 1 (JAK1)/ signal transducer and activator of transcription 1 (STAT1) signaling pathways were detected through Western blotting.
RESULTS: JPQT treatment decreased the lipid levels of triglyceride, low-density lipoprotein, and cholesterol, the inflammatory cytokine levels of interleukin 1 beta (IL-1β), IL-6 and IL-8 in rat serum, but increased high-density lipoprotein and IL-10 serum levels. JPQT treatment ameliorated pathological changes in the aorta of AS model rats. Moreover, JPQT upregulated SOCS1 protein expression and down-regulated phosphorylated protein expression levels of p-JAK1 and p-STAT1.
CONCLUSION: These results suggest that JPQT induces anti-atherosclerosis effects through anti-inflammatory and inhibiting JAK/STAT signaling pathways in HFD fed rats.

Key words: anti-inflammatory agents, Janus kinases, STAT transcription factors, signal transduction, diet, high-fat, antiatherosclerosis, Jianpi Qutan Fang

LIU Yue, ZHANG Fan, HAN Xiaomeng, XU Ningyang, ZHAO Yu, WANG Qige, WANG Jianan, LU Bingjiu, Zhang Yan. Jianpi Qutan Fang (健脾祛痰方) induces anti-atherosclerosis and ameliorates endothelial cell injury in high-fat diet rats via an anti-inflammatory and inhibiting Janus kinase/signal transducer and activator of transcription signaling pathway[J]. Journal of Traditional Chinese Medicine, 2023, 43(6): 1168-1175.

Figures/Tables 6

Table 1 Effect of JPQT on rat serum lipid levels (μmol/L, $\bar{x}$ ± s)

Group	n	TC	TG	LDL	HDL
Normal	6	198.5±34.7^a	36.7±7.3^a	32.0±4.5^a	73.6±4.6^a
Model	6	569.4±56.5^b	103.1±3.2^b	90.4±9.6^b	17.9±3.1^b
Atorvastatin	6	438.0±27.1^ab	90.0±4.6^ab	64.3±5.7^ab	49.5±2.6^ab
Quarter-dose JPQT	6	510.7±28.3^ac	102.2±3.0^d	87.0±3.0^d	32.5±1.8^ab
Half-dose JPQT	6	467.2±21.8^ab	97.5±1.7^d	76.6±2.4^ab	33.4±2.8^ab
JPQT	6	367.2±43.5^ab	90.6±3.7^ab	63.6±5.6^ab	48.5±3.2^ab
Double-dose JPQT	6	321.1±42.9^ab	76.4±7.1^ab	53.5±4.6^ab	61.4±2.8^ab
Quadruple-dose JPQT	6	208.2±38.8^a	56.1±6.1^ab	46.9±2.7^ab	70.3±5.3^a
F value		76.395	137.444	37.491	194.175
P value		<0.001	<0.001	<0.001	<0.001

Table 2 Effect of JPQT on rat serum inflammatory cytokines levels ($\bar{x}$ ± s)

Group	n	IL-1β (ng/L）	IL-6 (pg/mL）	IL-8 (ng/L）	IL-10 (ng/L）
Normal	6	12.8±1.8^a	26.9±2.1^a	138.3±6.8^a	47.4±2.1^a
Model	6	29.8±0.9^b	76.0±1.6^b	317.1±9.1^b	18.3±1.5^b
Atorvastatin	6	22.0±0.5^ab	56.0±1.7^ab	253.1±18.1^ab	41.2±1.9^ab
Quarter-dose JPQT	6	27.1±1.5^ab	68.4±1.8^ab	284.5±11.8^ab	28.6±2.1^ab
Half-dose JPQT	6	23.5±1.2^ab	63.6±1.8^ab	284.0±6.7^ab	32.3±2.2^ab
JPQT	6	21.7±0.5^ab	55.4±2.5^ab	249.9±9.4^ab	38.5±1.1^ab
Double-dose JPQT	6	19.5±0.7^ab	47.2±2.0^ab	203.4±14.9^ab	41.3±1.4^ab
Quadruple-dose JPQT	6	18.5±1.2^ab	40.3±3.7^ab	174.4±8.4^ab	46.2±1.2^ab
F value		127.489	296.516	174.695	131.482
P value		<0.001	<0.001	<0.001	<0.001

Table 3 Measurement of mitochondrial respiratory enzyme activity (U/g, $\bar{x}$ ± s)

Group	n	Complex I	Complex Ⅱ	Complex Ⅲ	Complex Ⅳ
Normal	6	6375±261^a	1265±202^a	1011±63^a	4608±215^a
Model	6	1618±407^b	199±38^b	231±27^b	555±70^b
Atorvastatin	6	4848±164^ab	762±44^ab	516±26^ab	2839±153^ab
Quarter-dose JPQT	6	3207±193^ab	257±29^b	323±20^ab	1003±158^ab
Half-dose JPQT	6	3992±241^ab	609±13^ab	354±29^ab	1561±169^ab
JPQT	6	4572±145^ab	687±81^ab	452±23^ab	2351±130^ab
Double-dose JPQT	6	5186±137^ab	944±55^ab	647±34^ab	3150±144^ab
Quadruple-dose JPQT	6	5918±233^ab	1029±44^ab	873±34^ab	3763±249^ab
F value		249.455	114.437	380.62	406.357
P value		<0.001	<0.001	<0.001	<0.001

Figure 1 JPQT ameliorated histopathological changes in rat aorta (HE staining, ×200) A: normal control group (normal diet + normal saline); B: model group (HFD + normal saline); C: Atorvastatin group (8 weeks, HFD + 10 mg/kg Atorvastatin); D: Quarter-dose Jianpi Qutan Fang (8 weeks, HFD + 2.225 g/kg Jianpi Qutan Fang); E: Half-dose Jianpi Qutan Fang (8 weeks, HFD + 4.45 g/kg Jianpi Qutan Fang); F: Jianpi Qutan Fang (8 weeks, HFD + 8.9 g/kg Jianpi Qutan Fang); G: Double-dose Jianpi Qutan Fang (8 weeks, HFD + 17.8 g/kg Jianpi Qutan Fang); H: Quadruple-dose Jianpi Qutan Fang (8 weeks, HFD + 35.6 g/kg Jianpi Qutan Fang). JPQT: Jianpi Qutan Fang; HE: hematoxylin-eosin staining.

Figure 2 JPQT ameliorated ultrastructural changes in rat aorta Transmission electron microscope observe the subcellular structure of the thoracic aorta. A-B: normal control group (normal diet + normal saline), C-D: model group (HFD + normal saline), E: atorvastatin group (8 weeks, HFD + 10 mg/kg Atorvastatin), F: JPQT group (8 weeks, HFD + 8.9 g/kg JPQT), G: double-dose JPQT group (8 weeks, HFD + 17.8 g/kg JPQT), H: quadruple-dose JPQT group (8 weeks, HFD + 35.6 g/kg JPQT). Scale bar = 2 μm. Red arrow: the endothelial cells; blue arrow: the blood cells; pink arrow: elastic fibers and collagen fibers; green arrow: mitochondria.

Figure 3 JPQT inhibited JAK/STAT signaling pathway via upregulating SOCS1 protein expression in the aortic tissues of HFD fed rat A: expression of aortic SOCS1/JAK1/STAT1 signaling pathway proteins levels by Western blot; B: normalized against GAPDH. 1: Normal: normal control group (normal diet + normal saline); 2: Model: model group (HFD + normal saline); 3: Atorvastatin: atorvastatin group (HFD + 10 mg/kg Atorvastatin); 4: Quarter-dose JPQT: quarter-dose Jianpi Qutan Fang (HFD + 2.225 g/kg JPQT); 5: Half-dose JPQT: half-dose Jianpi Qutan Fang (HFD + 4.45 g/kg JPQT); 6: JPQT: Jianpi Qutan Fang (HFD + 8.9 g/kg JPQT); 7: Double-dose JPQT: double-doseJianpi Qutan Fang (HFD + 17.8 g/kg JPQT); 8: Quadruple-dose JPQT: quadruple-dose Jianpi Qutan Fang (HFD + 35.6 g/kg JPQT). JPQT: Jianpi Qutan Fang; JAK: Janus kinase; STAT: signal transducer and activator of transcription; SOCS1: suppressor of cytokine signaling 1; HFD: high-fat diet. Student t-test and one-way analysis of variance were used for comparison analysis. aP < 0.01 compared with model group; bP < 0. 01 compared with the normal group; cP < 0.05 compared with model group; dP < 0.05 compared with the normal group. Statistical significance between the groups was determined by one-way analysis of variance. The sample size for each group was 5.

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