Moghadam Fariborz, Hajian Mehdi, Rouhollahi Varnosfaderani Shiva, Jafarpour Farnoosh, Nasr Esfahani Mohammad Hossein
Department of Animal Biotechnology, Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran; Islamic Azad University, Tonekabon, Iran.
Department of Animal Biotechnology, Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
Theriogenology. 2021 Feb;161:57-64. doi: 10.1016/j.theriogenology.2020.11.022. Epub 2020 Nov 30.
Lipopolysaccharide (LPS) significantly reduces pre- and post-implantation developmental competence of embryos. One of the reason of this effect could be a consequence of TLR4-mediated inflammation. In this study, we assessed the anti-inflammatory effect of peroxisome proliferator activated receptor γ (PPAR γ) agonist, rosiglitazone (RGZ), in LPS-treated mouse embryos. Initially, the optimal doses of LPS, RGZ and GW9662 (a potent and selective PPARγ antagonist) were determined by treating the mouse zygotes up to blastocyst stage and assessment of compaction and blastocyst rates. Quantitative PCR was used to assess the mRNA expression of inflammatory cytokines. Immunostaining was used to study the translocation of PPARγ in blastocysts. Finally, the blastocysts were transferred to surrogate mouse to determine the post-implantation developmental competence. 0.0625 mg/mL of LPS significantly reduced the developmental competency by around 50% compared to control group. 10 μM of RGZ significantly ameliorated the toxic effect of LPS, which was also significantly reversed by 1.25 μM GW9662. Through immunostaining, it was shown that LPS could prevent the translocation of PPARγ to nucleus; and translocation was facilitated by RGZ and this effect was reversed by GW9662. A similar effect was also observed for the mRNA expression of inflammatory cytokines (Il-1β and Il-6). LPS significantly increased the expression of these cytokines, while RGZ significantly reduced their expression, which was also significantly reversed by GW9662. It was also shown that embryos exposed to LPS had significantly reduced post implantation developmental competence which was considerably improved by treatment with RGZ. In conclusion, these data may have clinical implications for ameliorating the adverse effects of LPS in dairy farming and infertility treatment.
脂多糖（LPS）显著降低胚胎植入前和植入后的发育能力。这种效应的原因之一可能是TLR4介导的炎症反应的结果。在本研究中，我们评估了过氧化物酶体增殖物激活受体γ（PPARγ）激动剂罗格列酮（RGZ）对LPS处理的小鼠胚胎的抗炎作用。首先，通过将小鼠受精卵培养至囊胚阶段并评估致密化和囊胚率，确定LPS、RGZ和GW9662（一种强效且选择性的PPARγ拮抗剂）的最佳剂量。采用定量PCR评估炎性细胞因子的mRNA表达。免疫染色用于研究PPARγ在囊胚中的转位。最后，将囊胚移植到代孕小鼠体内，以确定植入后的发育能力。与对照组相比，0.0625mg/mL的LPS显著降低了发育能力约50%。10μM的RGZ显著改善了LPS的毒性作用，而1.25μM的GW9662也显著逆转了这种作用。通过免疫染色表明，LPS可阻止PPARγ转位至细胞核；RGZ促进了转位，而GW9662逆转了这种作用。炎性细胞因子（Il-1β和Il-6）的mRNA表达也观察到类似的效果。LPS显著增加了这些细胞因子的表达，而RGZ显著降低了它们的表达，GW9662也显著逆转了这种作用。还表明，暴露于LPS的胚胎植入后发育能力显著降低，而RGZ处理可显著改善这种情况。总之，这些数据可能对改善LPS在奶牛养殖和不孕症治疗中的不良反应具有临床意义。
