Huangqin Qingrechubi Capsule alleviates inflammation and uric acid and lipid metabolism imbalance in rats with gouty arthritis by inhibiting the PTEN/PI3K/AKT signaling pathway

doi:10.12122/j.issn.1673-4254.2024.08.03

Abstract

Abstract:

Objective To investigate the effects of Huangqin Qingrechubi Capsule (HQC) on inflammation and uric acid and lipid metabolism in rats with gouty arthritis (GA) and its mechanism. Methods SD rat models of GA established by injecting monosodium urate into the right ankle joint were treated with saline, colchicine and HQC at low, medium and high doses (n=10) by gavage for 7 days. Toe swelling of the rats was detected at 4, 8, 24, 48 and 72 h after modeling, and synovial histological changes were observed with HE staining. Serum levels of interleukin-10 (IL-10), IL-18, tumor necrosis factor‑α (TNF‑α), transforming growth factor-β1 (TGF-β1), adiponectin, leptin, resistin and visfatin were measured by ELISA, and the levels of high-density lipoprotein cholesterol (HDL-C), triglyceride (TG), total cholesterol (TC), and uric acid (BUA) were detected. RT-qPCR and Western blotting were used to detect the mRNA expressions of phosphatase and tensin homolog (PTEN), phosphatidylinositol-3-kinase (PI3K) and protein kinase B (AKT) and the protein expressions of PTEN, PI3K, p-PI3K, AKT and p-AKT. Results The rat models of GA showed obvious toe swelling, which reached the peak level at 48 h. HE staining revealed massive inflammatory cell infiltration and synovial tissue hyperplasia. The rat models showed significantly increased expressions of TNF-α, TGF-β1, IL-18, TC, TG, leptin, resistin and visfatin, BUA, p-PI3K, and p-AKT and lowered levels of IL-10, APN, HDL-C, and PTEN. Treatment with HQC and colchicine obviously improved these changes and alleviated synovial pathologies and toe swelling in the rat models. Conclusion HQC can improve inflammation and correct the imbalance of uric acid and lipid metabolism in GA rats possibly by inhibiting the PTEN/PI3K/AKT signaling pathway.

Key words: gouty arthritis, Huangqin Qingrechubi Capsule, inflammation, uric acid metabolism, lipid metabolism, PTEN/PI3K/AKT signaling pathway

Xianheng ZHANG, Jian LIU, Qi HAN, Yiming CHEN, Xiang DING, Xiaolu CHEN. Huangqin Qingrechubi Capsule alleviates inflammation and uric acid and lipid metabolism imbalance in rats with gouty arthritis by inhibiting the PTEN/PI3K/AKT signaling pathway[J]. Journal of Southern Medical University, 2024, 44(8): 1450-1458.

Figures/Tables 8

Tab.1 Primer design table

Gene	Amplicon size	Primer sequence (5'→3')
PTEN	140	F:TATCTTGTGCTCACCCTGAC R:CTGCTAGCCTCTGGATTTGA
PI3K	123	F:CGAAACAAAGCCGAGAACCT R:GACGCAATGTTTGACTTCGC
AKT	143	F:CAGGTTCACCCAGTGACAAC R:CTCCTTCACCAGGATCACCT
β-actin	150	F:CCCATCTATGAGGGTTACGC R:TTTAATGTCACGCACGATTTC

Tab.2 Antibody information

Antibody	Dilution rate	Species
PI3K	1:1000	Rabbit
p-PI3K	1:5000	Mouse
AKT	1:1000	Rabbit
p-AKT	1:1000	Rabbit
PTEN	1:1000	Rabbit
GAPDH	1:2000	Mouse

Fig.1 Right ankle joints of the rats in each group after MSU injection 48 h. A: Normal control group; B: Model group; C: HQC-L group; D: HQC-M group; E: HQC-H group; F: Colchicine group.

Tab.3 Effect of HQC on toe swelling in GA rats (Mean±SD, mL, n=10)

Groups	4 h	8 h	24 h	48 h	72 h
Normal	1.856±0.025	1.796±0.023	1.702±0.027	1.650±0.025	1.645±0.029
Model	2.015±0.073*	2.156±0.045*	2.335±0.058*	2.547±0.059*	2.456±0.068*
HQC-L	1.945±0.059^△#	2.096±0.045^△#	2.170±0.059^△#	1.954±0.057^△#	1.741±0.065^△#
HQC-M	1.908±0.033^△#	1.995±0.036^△#	1.896±0.026^△#	1.805±0.040^△#	1.693±0.040^△#
HQC-H	1.896±0.038^△#	1.920±0.035^△	1.855±0.030^△	1.727±0.040^△	1.642±0.034^△
Colchicine	1.853±0.036^△	1.914±0.040^△	1.857±0.047^△	1.714±0.063^△	1.638±0.030^△

Fig.2 HE staining of synovial tissue of the rats in each group. Arrows indicate infiltration of inflammatory cells. A: Normal control group; B: Model group; C: HQC-L group; D: HQC-M group; E: HQC-H group; F: Colchicine group.

Fig.3 Comparison of TNF-α, TGF-β1, IL-18 and IL-10 expressions among the groups. *P<0.05 vs A; △P<0.05 vs B; #P<0.05 vs F. A: Normal control group; B: Model group; C: HQC-L group; D: HQC-M group; E: HQC-H group; F: Colchicine group.

Fig.4 Comparison of TC, TG, HDL-C, APN, LP, RS, VF and BUA levels among the groups. *P<0.05 vs A; △P<0.05 vs B; #P<0.05 vs F. A: Normal control group; B: Model group; C: HQC-L group; D: HQC-M group; E: HQC-H group; F: Colchicine group.

Fig.5 Comparison of mRNA expressions of PTEN, PI3K and AKT among the groups. *P<0.05 vs A; △P<0.05 vs B; #P<0.05 vs F. A: Normal control group; B: Model group; C: HQC-L group; D: HQC-M group; E: HQC-H group; F: Colchicine group.

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