Inflammatory regulation by restraining M2 microglial polarization: Neurodestructive effects of Kallikrein-related peptidase 8 activation in intracerebral hemorrhage.
Inflammatory regulation by restraining M2 microglial polarization: Neurodestructive effects of Kallikrein-related peptidase 8 activation in intracerebral hemorrhage.
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DOI：
10.1016/j.intimp.2023.110855
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发表时间：
2023-09
影响因子：
5.6
通讯作者：
Ling Tang;Liyuan Wang;Feng Jin;Yuehan Hao;Tianming Zhao;Wen-Fei Zheng;Zhiyi He
中科院分区：
文献类型：
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作者：
Ling Tang;Liyuan Wang;Feng Jin;Yuehan Hao;Tianming Zhao;Wen-Fei Zheng;Zhiyi He
脑出血是一种脑血管疾病。kallikrein相关肽酶8 （KLK8）是一种丝氨酸肽酶，其在脑出血中的作用尚不清楚。Western blot （WB）结果显示，自体血液注射24 h后，KLK8在大鼠血肿周围组织中表达上调。KLK8过表达加重了大鼠的行为缺陷，增加了脑组织含水量和氟玉B （FJB）阳性神经元数量。免疫荧光（IF）检测显示，过表达的klk8可促进大鼠血肿周围组织中Iba-1和iNOS的表达。WB和ELISA证实，过表达的klk8增加了大鼠血肿周围组织中COX-2、iNOS和Arg-1的表达以及IL-6、IL-1β和TNF-α的含量。IF染色证实CCR5与Iba-1共表达，WB结果显示CCR5表达升高，p-PKA和p-CREB表达降低。给药Maraviroc （MVC， CCR5抑制剂）可挽救klk8诱导的大鼠行为缺陷和脑损伤（水含量和fjb阳性神经元数量减少）。此外，MVC还能抑制过表达的klk8诱导的p-PKA和p-CREB的表达以及血周组织中IL-6、IL-1β和TNF-α的含量。Co-IP证实了CCR5和CCL14在HMC3细胞中的结合。Transwell实验表明，KLK8 + CCL4促进了细胞的趋化活性，这是由MVC拯救的。在HMC3细胞中应用MVC也证实了KLK8/CCL14/CCR5轴在ICH损伤中的生物学功能。总之，我们的研究表明，KLK8过表达加剧了脑出血过程，并参与了小胶质细胞的激活。KLK8可能激活CCL14从而开启下游的CCR5/PKA/CREB通路，为今后的治疗提供理论依据。
Intracerebral hemorrhage (ICH) is a cerebrovascular disease. Kallikrein-related peptidase 8 (KLK8) is a serine peptidase, while its role in ICH remains unclarified. Western blot (WB) showed that KLK8 was upregulated in rat perihematomal tissues 24 h following autologous blood injection. KLK8 overexpression aggravated behavioral deficits and increased water content and Fluoro-Jade B (FJB)-positive neuron numbers in brain tissue of rats. Immunofluorescence (IF) assay showed that overexpressed-KLK8 promoted Iba-1 and iNOS expression in perihematomal tissue of rats. Overexpressed-KLK8 increased COX-2, iNOS, and Arg-1 expression and the content of IL-6, IL-1β, and TNF-α in perihematomal tissue of rats, confirmed by WB and ELISA. IF staining confirmed the expression of CCR5 was co-expressed with Iba-1, and the WB results shown increased CCR5 expression and decreased p-PKA and p-CREB expression in perihematomal tissue. Maraviroc (MVC, CCR5 inhibitor) administration rescued KLK8-induced behavioral deficits and brain injury (decreased water content and FJB-positive neuron numbers) in rats. Additionally, MVC suppressed p-PKA and p-CREB expression and the content of IL-6, IL-1β, and TNF-α in perihematomal tissue, induced by overexpressed-KLK8. Co-IP confirmed the binding of CCR5 and CCL14 in HMC3 cells. Transwell assay shown that KLK8 plus CCL4 promoted the chemotactic activity of cells, which was rescued by MVC. The biological function of KLK8/CCL14/CCR5 axis in ICH injury was also proved by MVC administration in HMC3 cells. Overall, our work revealed that KLK8 overexpression aggravated ICH process and involved in microglial activation. KLK8 might activate CCL14 thereby turning on downstream CCR5/PKA/CREB pathway, providing a theoretical basis for future therapy.