Gupta Mahesh, Bahiram Krushna Bibichand, Sardar Vikas Muralidhar, Korde Jayant Pandurang, Magar Swapnil P, Bonde Sachin Wasudeorao, Kurkure Nitin Vasantrao
Department of Veterinary Physiology, Nagpur Veterinary College, Nagpur, India.
Department of Veterinary Biochemistry, Nagpur Veterinary College, Nagpur, India.
Reprod Domest Anim. 2019 Sep;54(9):1291-1303. doi: 10.1111/rda.13529. Epub 2019 Aug 10.
Adiponectin is an adipocyte-derived hormone regulating energy metabolism, insulin sensitivity and recently found to regulate reproduction. The current study was carried out to investigate gene and protein expression, immunolocalization of adiponectin and its receptors AdipoR1 and AdipoR2 in ovarian follicles of different developmental stages in water buffalo (Bubalus bubalis) and to investigate the effect of adiponectin on steroid production in cultured bubaline granulosa cells. qPCR, western blotting and immunohistochemistry were applied to demonstrate mRNA expression, protein expression and immunolocalization, respectively. The results indicate that adiponectin, AdipoR1 and AdipoR2 were present in granulosa cells (GC) and theca interna (TI) of ovarian follicles and the expression of adiponectin, AdipoR1, AdipoR2 in GC and AdipoR1 and AdipoR2 in TI increased with increase in follicle size (p < .05). Expression of adiponectin was high in small and medium size follicles in TI. The adiponectin and its receptors were immunolocalized in the cytoplasm of GC and TI cells. Further, in the in-vitro study, GCs were cultured and treated with recombinant adiponectin each at 0, 1 and 10 µg/ml alone or with follicle stimulating hormone (FSH) at 30 ng/ml) or Insulin-like growth factor I (IGF-I) at 10 ng/ml for 48 hr after obtaining 75%-80%s confluency. Adiponectin at 10 µg/ml increased IGF-I-induced estradiol (E ) and progesterone (P ) secretion and FSH-induced E secretion from GC and also increased the abundance of factors involved in E and P production (cytochrome P45019A1 [CYP19A1] and 3-beta-hydroxysteroid dehydrogenase [3β-HSD]). In conclusion, this study provides novel evidence for the presence of adiponectin and its receptors in ovarian follicles and modulatory role of adiponectin on steroid production in buffalo.
脂联素是一种由脂肪细胞分泌的激素，可调节能量代谢、胰岛素敏感性，最近还发现其能调节生殖功能。本研究旨在调查水牛（Bubalus bubalis）不同发育阶段卵泡中脂联素及其受体AdipoR1和AdipoR2的基因和蛋白表达、免疫定位，并研究脂联素对培养的水牛颗粒细胞类固醇生成的影响。分别应用qPCR、蛋白质印迹法和免疫组织化学来证明mRNA表达、蛋白质表达和免疫定位。结果表明，脂联素、AdipoR1和AdipoR2存在于卵泡的颗粒细胞（GC）和内膜细胞（TI）中，GC中脂联素、AdipoR1、AdipoR2以及TI中AdipoR1和AdipoR2的表达随卵泡大小增加而升高（p < 0.05）。TI中小卵泡和中等大小卵泡中脂联素表达较高。脂联素及其受体在GC和TI细胞的细胞质中进行免疫定位。此外，在体外研究中，培养GC并在达到75%-80%汇合度后，分别用0、1和10 μg/ml的重组脂联素单独处理，或与30 ng/ml的促卵泡激素（FSH）或10 ng/ml的胰岛素样生长因子I（IGF-I）共同处理48小时。10 μg/ml的脂联素增加了IGF-I诱导的颗粒细胞雌二醇（E）和孕酮（P）分泌以及FSH诱导的颗粒细胞E分泌，还增加了参与E和P生成的因子（细胞色素P45019A1 [CYP19A1]和3-β-羟基类固醇脱氢酶[3β-HSD]）的丰度。总之，本研究为水牛卵泡中脂联素及其受体的存在以及脂联素对类固醇生成的调节作用提供了新证据。
