Journal of Jiangsu University(Medicine Edition).
2024, 34(06):
476-484.
Objective: To explore the effect of CXCL14 on pyroptosis of adipocytes and the formation of atherosclerosis in diabetic microenvironment. Methods: ① ApoE-/- mice were intraperitoneally injected with streptozotocin to construct diabetes mellitus, and diabetic ApoE-/- mice were fed with high fat diet for 20 weeks, and the diabetic atherosclerosis model (model group) was constructed, and control mice were only fed on a high-fat diet (AS group); ② Anti-CXCL14 short peptide was injected subcutaneously on the medial hind limb of diabetic mice (anti-CXCL14 group), and control diabetic mice were injected subcutaneously with normal saline only; ③ Adeno-associated virus (AAV) was injected in situ into posterior inguinal subcutaneous adipose tissue in diabetic mice to inhibit gastrointestinal dermatin (GSDMD)mediated pyroptosis, divided into: AAV-shscramble group, AAV-shGSDMD group, anti-CXCL14+AAV-shscramble group, anti-CXCL14+AAV-shGSDMDgroup. After 20 weeks of high-fat diet of the mice, the serum of the mice was collected under anesthesia, and the blood lipid levels (total cholesterol, triglycerides, LDL-C, and HDL-C) of the mice were analyzed by biochemical detection kit. The mice were sacrificed, and the epididymal adipose tissue was scanned by electron microscopy to observe the changes of fat cells. qRT-PCR was used to analyze the changes of pyroptosisrelated factors GSDMD, aspartate proteolytic enzyme-1 (Caspase-1), NLR family pyridine domain protein 3 (NLRP3) inflammasome, interleukin-1β (IL-1β) and inflammatory factor interleukin-6 (IL-6). The mouse aorta was isolated and extracted, and the relative area of atherosclerotic plaques was observed by HE staining and oil red O staining. Results: Compared with the AS group, the hypertrophy and number of adipocytes of adipose tissue in the epididymis in the model group decreased (P<0.01), the size of aorticplaque increased, the levels of total cholesterol, triacylglycerol and LDL-C were significantly increased, and HDL-C was significantly reduced. The levels of pyroptosisrelated factors and IL-6 in adipose tissue of epididymis were significantly increased (P<0.05), indicating that diabetes promoted pyroptosis of AS plaque and adipose tissue in mice. Injection of anti-CXCL14 short peptide could reduce the size of aortic plaque, improve blood lipid levels, and inhibit adipose tissue pyroptosis, which increases the number of fat cells. After GSDMD knockdown, the number of adipocytes increased and the area of aortic plaques decreased. However, after the injection of anti-CXCL14 immune peptide, there was no significant change in atherosclerosis in the AAV-shGSDMDgroup. Conclusion: Anti-CXCL14 can attenuate adipose tissue pyroptosis and alleviate the development of diabetic atherosclerosis.
[Key words]CXCL14; pyroptosis; adipose tissue; atherosclerosis; diabetic microenvironment