Dual role of osteoblastic proenkephalin derived peptides in skeletal tissues

Proenkephalin encodes a group of small peptides with opiate-like activity, the endogenous opioids, known to function as neurohormones, neuromodulators, and neurotransmitters. Recently, we have demonstrated that in addition to its abundance in fetal brain tissue, proenkephalin is highly expressed in nondifferentiated mesodermal cells of developing fetuses. We identified the skeletal tissues, bone, and cartilage as major sites of proenkephalin expression. To examine the possibility that proenkephalin is involved in bone development we have studied the expression of this gene in bone-derived cells, its modulation by bone active hormones, and the effects of enkephalin-derived peptides on osteoblastic phenotype. Our studies revealed that osteoblastic cells synthesize high levels of proenkephalin mRNA which are translated, and the derived peptides are secreted. Reciprocal interrelationships between osteoblast maturation and proenkephalin expression were established. These results together with our observations demonstrating inhibitory effects of proenkephalin-derived peptides on osteoblastic alkaline phosphatase activity, strongly support the notion that proenkephalin is involved in bone development. A different direction of research by other investigators has established the capability of the opioid system in the periphery to participate in the control of pain. On the basis of these two lines of observation, we would like to present the following hypothesis: The potential of embryonic skeletal tissue to synthesize proenkephalin-derived peptides is retained in the adult in small defined undifferentiated cell populations. This potential is realized in certain situations requiring rapid growth, such as remodeling or fracture repair. We suggest that in these processes, similarly to the situation in the embryo, the undifferentiated dividing cells produce the endogenous opioids. In the adult these peptides may have a dual function, namely participating in the control of tissue regeneration and in the control of pain. © 1994 Wiley-Liss, Inc.     

压脚痛阈测定法的改进和应用

本文介绍一种改进的压脚测痛法。采用电机驱动替代用手挤压橡皮球完成升压过程;通过一个自动的电路切断结构判定抽脚反应,较目视更加客观;用读数保持电路记录压力变化,较直接读取刻度值更加准确,可靠。这些优点,业经电针和吗啡镇痛实验验证。     

电针、吗啡镇痛和耐受时某些脑区线粒体结合钙的变化

采用 Tb~(3+)荧光探针和离子选择电极研究了电针和吗啡镇痛及镇痛耐受时,动物不同脑区游离 Ca~(2+)和线粒体膜结合 Ca~(2+)的变化。实验结果表明,电针和吗啡都有较强的镇痛作用,与此同时,导水管周围灰质和下丘脑的线粒体膜结合 Ca~(2+)升高。脑室内预注钌红,则能降低这两个脑区的线粒体膜结合 Ca~(2+)和痛阈。另一方面,在电针或吗啡耐受时,两脑区的游离 Ca~(2+)浓度增加,线粒体膜结合 Ca~(2+)降低。结果提示,神经细胞质膜内外 Ca~(2+)的移动可能在电针和吗啡镇痛中起某种调节作用。     

电针和吗啡对伤害性刺激猫内脏大神经引起的海马诱发电位的影响

在氯醛糖麻醉并箭毒化的50只猫中,观察了强电流刺激内脏大神经时的背侧海马诱发电位(Hippocampal evoked potential,HEP)以及电针和吗啡对其影响。结果显示,用20V电压刺激内脏大神经可以兴奋包括 Aδ和 C 纤维在内的几乎全部纤维,并且可以在海马内记录到波形稳定的 HEP。以负波为主的 HEP 在海马锥体细胞层附近以及下脚复合体的表浅层记录到,以正波为主的 HEP 在海马锥体细胞顶树突的中下部记录明显。只有刺激强度达到兴奋内脏大神经的 Aδ纤维时才可记录出 HEP。吗啡对 HEP 有显著的抑制作用。电针也可以明显抑制 HEP 的振幅,并有后作用。上述结果提示,内脏大神经中的感觉纤维,包括痛纤维的冲动可以传至海马。     

家兔中脑导水管周围灰质中甲七肽样免疫活性物质具有镇痛作用并参与电针镇痛

以辐射热照射家兔鼻嘴部皮肤,测定甩头反应潜伏期(ERL)作为痛阈。通过预先埋植的慢性套管向中脑导水管周围灰质(PAG)注射甲七肽降解酶抑制剂 Captopril,观察其镇痛作用以及加强电针镇痛的作用能否被特异的甲七肽抗血清所对抗。(1)单侧 PAG 注射 Captopril240 nmol 的镇痛作用可为同一部位注射甲七肽抗血清(1μl)所翻转,注入甲啡肽抗血清则无效。(2)单侧 PAG 注射 Captopril 60 nmol 有加强电针镇痛的作用。该作用可被1μl 甲七肽抗血清所完全取消,将抗血清量减少到0.1μl 则无效。以上结果说明 PAG 内的甲七肽样免疫活性物质在镇痛和电针镇痛中发挥重要作用。     

家兔中脑导水管周围灰质中注射八肽胆囊收缩素(CCK-8)拮抗吗啡镇痛和电针镇痛

家兔单侧PAG内注射CCK-83ng,能使静脉注射4mg/kg吗啡引起的镇痛作用降低73％或使电针镇痛效果降低67％。在1.5—6.0ng范围内呈量效关系。无硫的CCK-8无此作用。PAG内注射CCK受体拮抗剂proglumide 4μg可翻转CCK-8的抗吗啡镇痛作用。说明PAG部位注射外源性CCK-8可通过CCK受体对抗阿片镇痛。 PAG内注射CCK-8抗血清可显著增强静脉注射2mg/kg吗啡的镇痛效果。PAG内注射CCK抗血清本身也能引起痛阈轻度升高。说明PAG内有内源性的CCK-8发挥紧张性的抗阿片镇痛作用。     

福尔马林致痛提高大鼠中枢μ阿片受体密度及电针的加强作用

用放射自显影方法观察到;（１）大鼠脚掌注射福尔马林后,某些与镇痛有关的脑区如尾核头部、伏隔核、杏仁核、中央灰质、脚间核、中缝大核、脊髓背角等结构中μ阿片受体密度明显增加（Ｐ＜０．０５,Ｐ＜０．０１）;（２）给予电针抑制痛反应的大鼠,在其大部分上述结构及扣带回、隔区、视前内侧区、内膝体、上丘、中缝背核及中央上核受体密度明显增加;与福尔马林注射组相比,脚间核、中央灰质尾端腹外侧区、腰膨大背角的受体密度进一步增加。从而在受体水平支持伤害性刺激可以激活体内内阿片肽能活动,而电针可以加强这一活动的设想。     

电针引起脊髓P物质释放的频率依赖性

我室以往的研究表明,不同频率的电针可引起脊髓释放不同种类的阿片肽。本工作观察Ｐ物质（ＳＰ）释放的频率依赖性,电针频率选择２,４,８,１５,３０和１００Ｈｚ,分别收集电针期间及电针前后各３０ｍｉｎ的脊髓灌流液,通过放射免疫方法测定大鼠电针有效组和电针无效组Ｐ物质免疫活性（ＳＰ－ｉｒ）．结果如下：（１）电针有效组：２Ｈｚ引起ＳＰ－ｉｒ降低,与电针前相比,Ｐ＜０．０１;４Ｈｚ电针前后ＳＰ－ｉｒ比较,无统计学意义;８,１５,３０,１００Ｈｚ时ＳＰ－ｉｒ均增加（Ｐ＜０．０１）,其中１５Ｈｚ时ＳＰ增加最多（Ｐ＜０．００１）,表明刺激引起ＳＰ释放有频率依赖性。（２）电针无效组：不论应用何种频率,电针前后脊髓灌流液中ＳＰ－ｉｒ变化不大（均Ｐ＞０．０５）。提示,电针时脊髓液中ＳＰ含量变化与镇痛效果有密切关系。     

The Ethanol Extract of Syringa oblata Heartwood,a Mongolian Folk Medicine Containing Major Lignans,Exerts Analgesic and Sedative Effects on Mice

The heartwood of Syringa oblata Lindl. (SO) is one of Mongolian folk medicines to treat insomnia and pain, while its pharmacological evaluation and underlying mechanism remain unclear. In this study, the sedative effect of ethanol extract of SO (ESO) was evaluated with the locomotor activity test and the threshold dose of pentobarbital sodium-induced sleep test in mice, and the hot plate test, acetic acid-induced writhing test, and formalin test in mice were used to evaluate its analgesic effect. The underlying mechanism of ESO analgesia was explored by RT-PCR and western blot analysis, which is associated with the regulation of the NF-κB signaling pathway. Besides, the main constituents of ESO were characterized by LC/MS data analysis and comparison with isolated pure compounds. The current findings brought evidence for clinical application and further pharmacological and phytochemical studies on SO.     

催产素在脊髓水平对电针镇痛的影响

采用玻璃微电极胞外记录和脊髓表面给药的方法观察了催产素（ＯＴ）、抗催产素血清（ＡＯＴＳ）以及电针穴位对背角神经元伤害性诱发放电的影响。结果表明：电针穴位或脊髓表面施加ＯＴ可部分抑制脊髓背角神经元的伤害性诱发放电;在电针的基础上施加ＯＴ则明显加强电针的抑制效应;相反,用ＡＯＴＳ预处理后,电针的抑制作用放取消。提示ＯＴ在脊髓水平参与了对痛觉信息的调制,并与一定频率的针刺镇痛有关。