目的：探讨原儿茶酸通过抑制IL-6/JAK1/STAT5信号通路抑制颅内动脉瘤进展的分子机制。方法：构建颅内动脉瘤大鼠模型,分组为NC组(空白对照组)、颅内动脉瘤组(CA组)、颅内动脉瘤模型+原儿茶酸组(CA+PCA组),影像学检测各组颅内血管壁内径,HE染色检测各组血管内皮情况。ELISA检测各组外周血中IL-6、JAK1和STAT5的表达,RT-PCR和WB检测各组组织样本中IL-6、JAK1和STAT5的表达水平。结果：相较于NC组,从第8周开始,CA组的血管壁厚度明显增高(P＜0.05),而CA+PCA组的血管壁厚度较CA组降低。HE染色结果显示,相较于NC组,CA组的血管内皮中巨噬细胞增多,而CA+PCA较CA组血管内皮巨噬细胞减少。ELISA实验显示,相较于NC组,CA组的IL-6、JAK1和STAT5表达升高,而CA+PCA组中IL-6、JAK1和STAT5表达较CA组降低。RT-PCR和WB结果与ELISA结果相同(P＜0.05)。结论：原儿茶酸通过抑制IL-6/JAK1/STAT5信号通路可抑制颅内动脉瘤进展。
Objective:To investigate the molecular mechanism of protocatechuic acid inhibiting the progression of intracranial aneurysms by inhibiting IL-6/JAK1/STAT5 signaling pathway. Methods: The rat model of intracranial aneurysm was constructed and divided into NC group (blank control group), intracranial aneurysm group (CA group) and intracranial aneurysm model+protocatechuic acid group (CA+PCA group). The inner diameter of intracranial vascular wall in each group was detected by imaging, and the vascular endothelium in each group was detected by HE staining. The expression of IL-6, JAK1 and STAT5 in peripheral blood of each group was detected by ELISA, and the expression levels of IL-6, JAK1 and STAT5 in tissue samples of each group were detected by RT-PCR and WB. Results: Compared with the NC group, the thickness of the vascular wall in the CA group was significantly increased from the eighth week (P＜0.05), while the thickness of the vascular wall in the CA+PCA group was lower than that in the CA group. The results of HE staining showed that compared with the NC group, the number of macrophages in the vascular endothelium of the CA group increased, while the number of macrophages in the vascular endothelium of the CA+PCA group decreased compared with the CA group. ELISA experiments showed that the expression of IL-6, JAK1 and STAT5 in CA group was higher than that in NC group, while the expression of IL-6, JAK1 and STAT5 in CA+PCA group was lower than that in CA group. RT-PCR and WB results were the same as ELISA results (P＜0.05). Conclusion: Protocatechuic acid inhibits the progression of intracranial aneurysms by inhibiting the IL-6/JAK1/STAT5 signaling pathway.
[1] Jin J, Duan J, Du L, et al. Inflammation and immune cell abnormalities in intracranial aneurysm subarachnoid hemorrhage(SAH): relevant signaling pathways and therapeutic strategies[J]. Front Immunol, 2022, 13: 1027756.
[2] Ajiboye N, Chalouhi N, Starke RM, et al. Unruptured cerebral aneurysms: evaluation and management[J]. Scientific World J, 2015, 2015: 954954.
[3] Fan H, Tian H, Jin F, et al. CypD induced ROS output promotes intracranial aneurysm formation and rupture by 8-OHdG/NLRP3/MMP9 pathway[J]. Redox Biol, 2023, 67: 102887.
[4] Ishiguro T, Furukawa H, Polen K, et al. Pharmacological inhibition of epidermal growth factor receptor prevents intracranial aneurysm rupture by reducing endoplasmic reticulum stress[J]. Hypertension, 2024, 81(3): 572-581.
[5] Ji H, Li Y, Sun H, et al. Decoding the cell atlas and inflammatory features of human intracranial aneurysm wall by Single-Cell RNA sequencing[J]. J Am Heart Assoc, 2024, 2024: e032456.
[6] Upadhyay P, Lali A. Protocatechuic acid production from lignin-associated phenolics[J]. Prep Biochem Biotechnol, 2021, 51(10): 979-984.
[7] He C, Yu L, Dan W, et al. Application of a simple microfluidic chip analysis technology to evaluate the inhibitory role of protocatechuic acid on shear-induced platelet aggregation[J]. Evid Based Complement Alternat Med, 2021: 5574413.
[8] Kakkar S, Bais S. A review on protocatechuic acid and its pharmacological potential[J]. ISRN Pharmacol, 2014, 2014: 952943.
[9] Bai L, Han X, Kee HJ, et al. Protocatechuic acid prevents isoproterenol-induced heart failure in mice by downregulating kynurenine-3-monooxygenase[J]. J Cell Mol Med, 2023, 27(16): 2290-2307.
[10] Frösen J, Cebral J, Robertson AM, et al. Flow-induced, inflammation-mediated arterial wall remodeling in the formation and progression of intracranial aneurysms[J]. Neurosurg Focus, 2019, 47(1): E21.
[11] Masella R, Santangelo C, D′Archivio M, et al. Protocatechuic acid and human disease prevention: biological activities and molecular mechanisms[J]. Curr Med Chem, 2012, 19(18): 2901-2917.
[12] Xiao G, Zhang M, Peng X, et al. Protocatechuic acid attenuates cerebral aneurysm formation and progression by inhibiting TNF-alpha/Nrf-2/NF-kB-mediated inflammatory mechanisms in experimental rats[J]. Open Life Sci, 2021, 16(1): 128-141.
[13] Al Olayan EM, Aloufi AS, Alamri OD, et al. Protocatechuic acid mitigates cadmium-induced neurotoxicity in rats: role of oxidative stress,inflammation and apoptosis[J]. Sci Total Environ, 2020, 723: 137969.
[14] Li L, Ma H, Zhang Y, et al. Protocatechuic acid reverses myocardial infarction mediated by β-adrenergic agonist via regulation of Nrf2/HO-1 pathway, inflammatory, apoptotic, and fibrotic events[J]. J Biochem Mol Toxicol, 2023, 37(3): e23270.
[15] Salama A, Elgohary R, Fahmy MI. Protocatechuic acid ameliorates lipopolysaccharide-induced kidney damage in mice via downregulation of TLR-4-mediated IKBKB/NF-κB and MAPK/Erk signaling pathways[J]. J Appl Toxicol, 2023, 43(8): 1119-1129.
[16] Kaewmool C, Kongtawelert P, Phitak T, et al. Protocatechuic acid inhibits inflammatory responses in LPS-activated BV2 microglia via regulating SIRT1/NF-κB pathway contributed to the suppression of microglial activation-induced PC12 cell apoptosis[J]. J Neuroimmunol, 2020, 341: 577164.
[17] Banerjee S, Biehl A, Gadina M, et al. JAK-STAT signaling as a target for inflammatory and autoimmune diseases: current and future prospects[J]. Drugs, 2017, 77(5): 521-546.
[18] Pang W, Hu F. C9ORF72 suppresses JAK-STAT mediated inflammation[J]. iScience, 2023, 26(5): 106579.
[19] Morelli M, Scarponi C, Mercurio L, et al. Selective immunomodulation of inflammatory pathways in keratinocytes by the janus kinase (JAK) inhibitor tofacitinib:implications for the employment of JAK-Targeting drugs in psoriasis[J]. J Immunol Res, 2018, 2018: 7897263.
[20] Valenzuela NM. JAKinibs prevent persistent, IFNγ-autonomous endothelial cell inflammation and immunogenicity[J]. Am J Physiol Cell Physiol, 2023, 325(1): C186-C207.
[21] Beckman JD, Dasilva A, Aronovich E, et al. JAK-STAT inhibition reduces endothelial prothrombotic activation and leukocyte-endothelial proadhesive interactions[J]. J Thromb Haemost, 2023, 21(5): 1366-1380.
[22] Johnson DE, O′Keefe RA, Grandis JR. Targeting the IL-6/JAK/STAT3 signalling axis in cancer[J]. Nat Rev Clin Oncol, 2018, 15(4): 234-248.
[23] Heinrich PC, Behrmann I, Haan S, et al. Principles of interleukin (IL)-6-type cytokine signalling and its regulation[J]. Biochem J, 2003, 374(Pt 1): 1-20.
[24] Kaur S, Bansal Y, Kumar R, et al. A panoramic review of IL-6:structure, pathophysiological roles and inhibitors[J]. Bioorg Med Chem, 2020, 28(5): 115327.
