ANTI-INFLAMMATORY STEROIDS AND COLLAGEN SYNTHESIS

抗炎类固醇用于各种皮肤病。 这些类固醇最明显的副作用之一是皮肤 萎缩 在过去的授予期间，我们已经表明，抗炎 类固醇选择性地减少I型前胶原的合成， 鸡皮肤和鸡中pro Alpha1（I）和pro Alpha2（I）的mRNA 皮肤成纤维细胞 我们现在建议确定 抗炎类固醇对细胞核中I型前胶原mRNA的影响， 用克隆的前胶原蛋白cDNA探针在细胞质和多聚核糖体中的表达 Alpha1（I）和pro Alpha2（I）mRNA。 糖皮质激素对糖尿病患者的影响 细胞质中I型前胶原mRNA的周转率将 测定 糖皮质激素对I型胶原基因转录的影响 还将测定前胶原核RNA。 原子核将被分离 来自对照和类固醇处理的成纤维细胞并在体外转录。 从对照成纤维细胞中分离的细胞核将与活化的 体外糖皮质激素受体与Ⅰ型前胶原转录 基因将被确定。 激活的效果 糖皮质激素受体对I型多聚腺苷酸化和剪接的影响 还将获取前胶原核RNA。 此外，来自 将测定对照和糖皮质激素处理的鸡皮肤成纤维细胞 聚（A）聚合酶活性。 我们将确定约束力的程度， 类固醇受体的5英尺前胶原克隆基因片段， 在体外成功转录。 的能力 调节I型前胶原合成的类固醇受体 将获得该基因片段的转录物。 我们还将确定 两种新的泼尼松龙酯衍生物， 甲基强的松龙和甲基20二氢强的松龙，以抑制 博来霉素诱导的鸡皮肤和鸡皮肤中的胶原合成 成纤维细胞 我们对这些新的类固醇感到特别兴奋， 它们具有抗炎作用，但不抑制正常皮肤胶原蛋白 合成. 这些研究将为糖皮质激素提供分子基础 抑制皮肤中前胶原合成。

Anti-inflammatory steroids are used in various dermatological diseases. One of the most conspicuous side effects of these steroids is skin atrophy. In the past granting periods we have shown that anti-inflammatory steroids selectively decrease type I procollagen synthesis and decrease mRNAs for both pro Alpha1(I) and pro Alpha2(I) in chick skin and in chick skin fibroblasts. We now propose to determine the effects of anti-inflammatory steroids on type I procollagen mRNA's in nuclei, in the cytoplasm and in polysomes by using cloned cDNA probes for procollagen pro Alpha1(I) and pro Alpha2(I) mRNA's. The effects of glucocorticoids on the turnover rates of type I procollagen mRNAs in the cytoplasm will be determined. The effect of glucocorticoid on the transcription of type I procollagen nuclear RNA will also be determined. Nuclei will be isolated from control and steroid treated fibroblasts and transcribed in vitro. Nuclei isolated from control fibroblasts will be incubated with activated glucocorticoid-receptor in vitro and transcription of type I procollagen genes will be determined. The effect of the activated glucocorticoid-receptor on polyadenylation and splicing of type I procollagen nuclear RNA will also be accessed. Furthermore, nuclei from control and glucocorticoid treated chick skin fibroblasts will be assayed for poly(A)polymerase activity. We will determine the degree of binding of the steroid-receptor to a 5 feet procollagen cloned gene fragment which has been successfully transcribed in vitro. The ability of the steroid-receptor to regulate the synthesis of procollagen type I transcripts by this gene fragment will be accessed. We will also determine the ability of two novel prednisolone ester derivatives, methylprednisolonate and methyl 20 dihydroprednisolonate, to inhibit bleomycin-induced collagen synthesis in chick skin and in chick skin fibroblasts. We are particularly excited about these new steroids since they are anti-inflammatory but do not inhibit normal skin collagen synthesis. These studies will provide a molecular basis for glucocorticoid inhibition of procollagen synthesis in skin.