Rahman Al-Nur Md Iftekhar, Yamashita Seiya, Islam Md Rashedul, Fujihara Taisuke, Yamaguchi Hayato, Kawahara Manabu, Takahashi Masashi, Takahashi Hideyuki, Gotoh Takafumi, Yamauchi Nobuhiko
Department of Animal and Marine Bioresource Sciences, Faculty of Agriculture, Kyushu University, Fukuoka, Japan.
Department of Animal Nutrition, Genetics and Breeding, Faculty of Animal Science and Veterinary Medicine, Sher-e-Bangla Agricultural University, Dhaka, Bangladesh.
Anim Sci J. 2020 Jan-Dec;91(1):e13350. doi: 10.1111/asj.13350.
This study investigated the effect of type-I interferon (IFN) on the expression of matrix metalloproteinases (MMPs) of the bovine endometrial stromal cells (BES) and epithelial cells (BEE). The cells were separated and purified from the caruncles and cultured in DMEM/F-12 containing 10% fetal bovine serum. Spheroids were generated by using ascorbate. Zymograms of the supernatant showed that BEE predominantly expressed MMP-9, whereas MMP-2 was expressed in BES and homo-spheroids. While MMPs expression was not detected in hetero-spheroids. Real-time quantitative PCR revealed that type-I IFN and P4 suppressed the gene expression of MMP-2 and MMP-9 in hetero-spheroids, respectively. On the other hand, gelatin zymography analysis of the supernatant showed that type-I IFN strongly promote the clearance of MMPs. While zymograms of the MMPs stocked in the hetero-spheroids were significantly reduced by type-I IFN. Phenylmethanesulfonyl fluoride and leupeptin (both are serine proteinase inhibitors) significantly repressed the clearance of MMP-2 and MMP-9 induced by type-I IFN. Moreover, collagen fibers in hetero-spheroids significantly decreased after the treatment with type-I IFN. In conclusion, it was suggested that type-I IFN participate in the tissue remodeling by regulation the clearance of MMPs.
本研究调查了I型干扰素（IFN）对牛子宫内膜基质细胞（BES）和上皮细胞（BEE）基质金属蛋白酶（MMPs）表达的影响。从肉阜中分离并纯化细胞，在含有10%胎牛血清的DMEM/F-12培养基中培养。通过使用抗坏血酸生成球体。上清液的酶谱显示，BEE主要表达MMP-9，而MMP-2在BES和同型球体中表达。而异型球体中未检测到MMPs表达。实时定量PCR显示，I型干扰素和孕酮分别抑制了异型球体中MMP-2和MMP-9的基因表达。另一方面，上清液的明胶酶谱分析显示，I型干扰素强烈促进MMPs的清除。而异型球体中储存的MMPs的酶谱被I型干扰素显著降低。苯甲基磺酰氟和亮抑酶肽（两者均为丝氨酸蛋白酶抑制剂）显著抑制了I型干扰素诱导的MMP-2和MMP-9的清除。此外，用I型干扰素处理后，异型球体中的胶原纤维显著减少。总之，提示I型干扰素通过调节MMPs的清除参与组织重塑。
