Zhuriheng Pills improve Adipose Tissue Dysfunction and Inflammation by modulating PPARγ to stabilize atherosclerotic plaques

Qiong Zhai¹Fangyuan Liang²Guanlin Yang³Zhimin Liu⁴Xin Dong¹Ren Bu¹Peifeng Xue¹Shengsang Na¹Xuan Zhang¹Pengwei Zhao¹Xiaoning Wang¹Qiang Wei¹Yuewu Wang^1*Jingkun Lu^1*

• ¹Inner Mongolia Medical University, Hohhot, China
• ²Nantong Vocational University, Nantong, Jiangsu Province, China
• ³Zaozhuang Thoracic hospital (Zaozhuang Cancer Hospital), Zaozhuang, China
• ⁴Chifeng College Affiliated Hospital, Chifeng, China

Background: Adipose tissue dysfunction and chronic inflammation contribute to atherosclerosis plaque development. ZRH is an effective Mongolian herbal formula used in treating coronary heart disease in China, but its mechanisms of action have not been fully elucidated. Purpose: To assess whether ZRH alleviates AS and stabilizes plaque, this study investigates the modulatory effects of ZRH on AS and adipose tissue profiles in atherosclerotic model mice with vulnerable plaque to reveal the potential mechanisms and representative Q-Markers of ZRH. Methods: In vivo, the vulnerable atherosclerotic plaque model was induced in ApoE-/- mice treated with intense co-stimulation. The anti-AS effect of ZRH was assessed by serum lipid profile, HE, Oil O red, Masson staining, IHC, immunofluorescence, and plasma lipidomics. In vitro, a co-culture model was established with 3T3-L1 adipocytes treated with PA and RAW264.7 macrophages treated with LPS. The potential mechanism and Q-Markers of ZRH were discovered by lipid content test, the inflammatory factors and adipocytokines analysis, flow cytometry for macrophage polarization, and Western blotting for PPARγ and UCP-1 proteins. Results: In vivo, ZRH stabilizes plaques by improving serum lipid profiles, lowering macrophage infiltration, and boosting collagen content in plaques. ZRH can counteract HFD-induced adipocyte hypertrophy, increase UCP-1 and PPARγ expression, enhance the "browning" of adipose tissue, and inhibit macrophage M1 polarization. Lipidomics results showed that the ZRH treatment increased the abundance of lipid species with multiple unsaturated bonds and decreased harmful TAG, DAG, and HexCer. In addition, ZRH regulates inflammatory factors and adipokines in co-culture to inhibit macrophage M1 polarization and adipocyte abnormal lipid metabolism. In contrast, RDK and its monomers have a stronger anti-inflammatory effect, whereas GZ and its monomers regulate lipid metabolism better. ZRH was shown to be a PPARγ agonist for improving adipose tissue dysfunction and inflammation for anti-AS effects of ZRH. MIX, which is made up of Ellagic acid, Quercetin, 3,3'-Di-O-methylellagic acid, Elemicin, and Safrole in equal proportions, is only one of ZRH's Q-markers. More research is needed on the roles of different ZRH metabolites. Conclusion: Our results demonstrated that ZRH stabilizes atherosclerotic plaques by ameliorating the adipose tissue dysfunction and inflammation via regulating the PPARγ pathway.