Expression and Cellular Localization of Interleukin-6 mRNA in Ovariectomized Rats

In order to observe the expression and cellular localization of interleukin-6 (IL-6) mRNA in bone tissue, ovaria of the rats were excised to develop osteoporosis model. The expression of IL-6 mRNA in bone tissues was detected by using dot blot hybridization assay and the cells producing IL6 identified and localized by using in situ hybridization respectively. The results showed that the expression of IL-6 mRNA was significantly increased in the ovariectomized rats as compared with that in normal control rats and strong IL-6 mRNA hybridization signals were detected in lining cells, osteoblasts and osteocytes. It was suggested that loss of ovarian function induced in vivo osteoblast lineage increased IL-6 mRNA expression. IL-6 might play important roles in the development of bone loss following ovariectomy.

癫痫患者脑组织TNF-αmRNA、IL-6mRNA以及HLA-Ⅱ抗原表达的改变

为探讨肿瘤坏死因子 (TNF-α)、白细胞介素 6 (1L - 6 )以及 HL A- 抗原表达在癫痫免疫学发病机制中的作用 ,采用地高辛标记 c DNA探针技术组织原位杂交方法研究癫痫患者 TNF- α m RNA和 IL- 6 m RNA在脑组织中的表达与分布 ;同时借助免疫组织化学技术检测癫痫患者脑组织中 HL A- 抗原的表达水平。结果发现 :癫痫患者脑组织中星形胶质细胞异常高表达 TNF- α m RNA和 IL- 6 m RNA;星形胶质细胞和小胶质细胞异常高表达 HL A- 类抗原。结果提示 :脑组织局部自分泌或旁分泌的细胞因子 (TNF-α和 IL - 6 )水平以及胶质细胞表面 HL A-

细胞因子(IL-6、TNF-α)在损伤时间中原位杂交法研究

采用原位杂交法，研究大鼠切削皮肤中细胞团子IL－6、TNF－αmRNA表达量，旨在探讨IL－6、TNF－α推断法医损伤时间的应用价值以及其在损伤生活反应中的分子机制。研究结果表明，根据大鼠损伤皮肤中TNF－αmRNA表达量能够区别生前伤与死后伤，并可以利用TNF－αmRNA表达量改变准确区别60min内和60min后损伤时间，但IL－6mRNA表达量在研究组内均未见阳性反应，不能推断损伤时间。

Naoxintong dose effects on inflammatory factor expression in the rat brain following focal cerebral ischemia

BACKGROUND： Certain components of tetramethylpyrazine, a traditional Chinese medicine, exhibit protective effects against brain injury. OBJECTIVE： To investigate the effects of different Naoxintong doses on expression of nuclear factor-kappa B （ kB）, interleukin-6, tumor necrosis factor-α, and complement 3 in rats following focal cerebral ischemia. DESIGN, TIME AND SETTING： The randomized experiment was performed at the Laboratory of Neurology, Second Hospital of Hebei Medical University from June 2004 to June 2006. MATERIALS： A total of 150 adult, healthy, male, Sprague Dawley rats, weighing 280-320g, were selected. Naoxintong powder （mainly comprising szechwan lovage rhizome, milkvetch root, danshen root, and radix angelicae sinensis） was obtained from Buchang Pharmacy Co., Ltd. in Xianyang City of Shanxi Province of China, lot number 040608. METHODS： The rats were randomly assigned into sham operation, saline, high-dose Naoxintong, moderate-dose Naoxintong, and low-dose Naoxintong groups, with 30 rats in each group. Rat models of middle cerebral artery occlusion were established using the suture method, with the exception of the sham operation group. Rats in the high-dose, moderate-dose and low-dose Naoxintong groups received 4, 2, and 1 g/kg Naoxintong respectively, by gavage. Rats in the saline group were treated with 1 mL saline by gavage All rats were administered by gavage at 5 and 23 hours following surgery, and subsequently, once per day. MAIN OUTCOME MEASURES： At 6, 24, 48, 72 hours, and 7 days following model establishment, brain water content was measured. Histopathological changes in brain tissues were detected using hematoxylin-eosin staining. Expression of nuclear factor- kB, interleukin-6, tumor necrosis factor- α, and complement 3 was examined by immunohistochemistry. RESULTS： A total of 150 rats were included in the final analysis with no loss. Brain water content was significantly increased in the ischemic hemisphere of rats from the saline, as well as the high-dose, moderate-dose, and low-dose Naoxintong groups at 24 hours, which reached a peak at 48 hours. At 6, 24, 48, 72 hours, and 7 days, brain water content was greater in the ischemic hemisphere of rats from the saline, as well as the high-dose, moderate-dose, and low-dose Naoxintong groups, compared with the sham operation group （P 〈 0.05）. At 24 and 48 hours, brain water content was reduced in the high-dose and moderate-dose Naoxintong groups, compared to the saline and low-dose Naoxintong groups （P 〈 0.05）. In the saline, as well as high-dose, moderate-dose, and low-dose Naoxintong groups, neuronal edema was observed at 6 hours surrounding the ischemic sites. Inflammatory cells appeared at 24 hours, reached a peak at 48 hours, and gradually diminished. A small amount of glial cell proliferation and neuronal degeneration were observed in the hippocampus at 72 hours following infarction. Microglial proliferation and aggregation were detected at 7 days after infarction. Expression of nuclear factor- kB, interleukin-6, tumor necrosis factor-α, and complement 3 was significantly less in the high-dose, moderate-dose, and low-dose Naoxintong groups, compared to the sham operation group （P 〈 0.05）. Expression of the above-mentioned inflammatory cytokines was lower in rat brain tissues of the high-dose Naoxintong group, compared to the low-dose Naoxintong group （P 〈 0.05）. CONCLUSION： High-dose Naoxintong and moderate-dose Naoxintong significantly alleviated rat brain edema and decreased expression of nuclear factor-kB, interleukin-6, tumor necrosis factor-α, and complement 3 in brain tissues. The protective effect of high-dose Naoxintong was most significant.

芪麝颈康丸对椎间盘细胞肿瘤坏死因子-α、白细胞介素-6mRNA表达的影响

目的:探讨芪麝颈康丸抑制颈椎间盘退变的分子机制。方法:将48只大鼠分成4组,切除其中36只(A、B、C 3组)颈后部浅、中层肌,造成颈椎间盘退变模型,D 组为正常假手术组。分别给予芪麝颈康丸(A)及对照药颈复康(B)等药物治疗,C、D 组不给药。4个月后将动物处死,采用原位杂交方法观察分析肿瘤坏死因子(TNF-α)、白细胞介素-6(IL-6)mRNA 在大鼠颈椎间盘组织中的表达规律及芪麝颈康丸对两者表达的影响,同时观察各组椎问盘退变程度的变化。结果:模型空白(C)组椎间盘退变明显,与 A、D 组比较。差异有显著性(P<0.05,P<0.01)。TNF-α mRNA 阳性信号各组均在椎间盘终板有较高表达,4组程度相似。除 C 组椎间盘髓核软骨样细胞中有明显表达外,其余各组髓核中未见 TNF-α mRNA 阳性信号表达。IL-6mRNA 阳性信号零散地见于各组椎间盘终板,且数量均很少。结论:TNF-α与颈椎间盘退变关系密切,而 IL-6在椎间盘退变过程中所起作用不大。芪麝颈康丸抑制髓核细胞 TNF-α mRNA 的表达可能是其疗效机制。

白细胞介素-6mRNA在脑肿瘤的表达及其意义

探讨ＩＬ－６与不同类型脑肿瘤的相关性。方法：采用地高辛标记ＩＬ－６ｃＤＮＡ探针，运用原位杂交技术检测ＩＬ－６ｍＲＮＡ在４０例脑肿瘤组织中的表达。结果：在良、恶性肿瘤组表达率分别为２３．１％、７０．４％，恶性肿瘤组明显高于良性组，两组差异非常显著（Ｐ＜０．０１）．星形细胞瘤低分化组表达率大子高分化组，两组差异显著（Ｐ＜０．０５）。结论：结果表明ＩＬ－６ｍＲＮＡ表达率与肿瘤性质及星形细胞瘤的病理分级有关。提示ＩＬ－６可能对胶质细胞瘤的生长增殖有促进作用，对高分化的星形细胞瘤进一步向恶性发展可能有关系，测定血清ＩＬ－６对恶性肿瘤的预后判断有一定意义。

Expression and significance of IL-6 mRNA and its protein in liver after injury in rats

In order to study the effects of IL-6 on the development of liver injury the distribution and cell localization of IL-6 mRNA and its protein in the liver was observed in rats after they were inflicted with 20%TBSA full-thickness burns and/or intraperitoneal injection of endotoxin (lipopolysaccharide, LPS). 156 Wistar rats were randomized into combined burn-endotixin group (CBEG), simple burn injury group (SBG),simple endotoxin group (SEG ) and normal control group (NCG). The changes of serum IL-6 level, and theexpression and localization of IL-6 and IL-6 mRNA in the liver were determined quantitatively, with imrnunohistochemistry (IHC) and in situ hybridization (ISH). It was found that serum IL-6 level showed 2 peak inthe first half hour and the 6th to 12th h after injury in CBEG and one peak in the 3rd h in SBG and in the 12thto 24th h in SEG. IL-6 was mainly located in the sinusoid endothelial cells and Kupffer cells and IL-6 mRNAwas distributed mainly in Kupffer cells, polymorphonuclear cells and macrophages. In addition, the expression levels of IL-6 and IL-6 mRNA were in agreement with the pathological changes of liver injury.It is concluded that IL-6 is one of the key cytokines in the pathogenesis of liver damages after combinedinjury of burns and endotoxin.

经小脑延髓池局部应用反义寡核苷酸对脑组织TNF-α表达的影响

目的 :观察 TNF-α反义寡核苷酸对 TNF-α的阻断作用。方法 :用 DNA合成仪合成 17聚体的 TNF-α正义寡核苷酸和反义寡核苷酸。 2 4只犬随机分为对照组和实验组 (包括人工脑脊液组、正义链组和反义链组 )。实验组动物用已建成的颅脑爆炸伤模型装置致伤 ,伤前经小脑 -延髓池分别注入人工脑脊液、TNF-α正义寡核苷酸和 TNF-α反义寡核苷酸 ,比较 3组动物脑组织 TNF-α m RNA表达的变化。结果 :与人工脑脊液组和正义链组相比 ,伤前经小脑 -延髓池脑内局部应用 TNF-α反义寡核苷酸组 ,颅脑爆炸伤后脑组织内 TNF- α m RNA表达量明显减少 (P<0 .0 1)。结论 :经小脑延髓池局部应用反义寡核苷酸可有效阻断颅脑爆炸伤后脑组织 TNF- α的表达。

Effects of Remifentanil Pretreatment on Inflammatory Response in Rats with Acute Cerebral Ischemia Injury

[Objectives]This study was conducted to investigate the effects of remifentanil pretreatment on inflammatory factors in rats with acute cerebral ischemia.[Methods]Sixty SD rats were randomly divided into the normal control group,sham operation group,ischemic brain injury group,and remifentanil pretreatment group.Except the normal control group,each group was divided into three subgroups(six in each group)according to the sampling time points of 6,12 and 24 h after execution.After modeling,the rats were scored for neurological deficit,and observed for pathological changes of neurons in the brain tissue by HE staining and the brain infarct volume by TTC staining,and the expression levels of TNF-α,IL-6 and IL-8 were detected by RT-PCR.[Results]HE staining:No significant changes were observed in the pathological morphology of the brain tissue in the blank group and sham operation group;and the neuronal structure of rats in the acute cerebral ischemia group was obviously damaged,and the neuronal damage in the remifentanil pretreatment group was less than that in the acute cerebral ischemia group at each time point.TTC staining:The gray brain infarct area in the remifentanil pretreatment group was significantly smaller than that in the cerebral ischemia group(P<0.05).RT-PCR detection results:The expression levels of TNF-α,IL-6 and IL-8 in the blank group and sham surgery group did not show significant changes at different times(P>0.05);and compared with the cerebral ischemia group,the expression levels of TNF-α,IL-6,and IL-8 in the remifentanil pretreatment group were significantly reduced at all time points(P<0.05).[Conclusions]Remifentanil pretreatment could protect the brain by reducing the expression of inflammatory factors after cerebral ischemia injury.

异种植骨局部肿瘤坏死因子-α和白细胞介素6 mRNA表达与意义

目的:观察肿瘤坏死因子-α(TNF-α)和白细胞介素6(IL-6)mRNA在异种植骨局部的表达情况并探讨其意义。材料与方法:将经过一定化学处理的牛松质骨载体植入小鼠股部肌袋内,于术后3、5、10、20和30天取材,制备石蜡切片,应用原位杂交技术检测植骨局部TNF-α和IL-6 mRNA的细胞定位表达并分析其表达规律。结果:术后3天植骨局部出现TNF-α mRNA阳性表达.术后5天出现IL-6mRNA阳性表达,TNF-α和IL-6 mRNA表达强度分别以术后10天和20天时为最强(P<0.01);TNF-α和IL-6 mRNA阳性细胞在载体周边呈散在性或局灶性分布,阳性信号分别定位于多种类型细胞的胞核内。结论:TNF-α和IL-6 mRNA在异种植骨局部多种类型细胞内出现阳性表达,提示这两种细胞因子可能是异种植骨局部细胞免疫及骨代谢调节的重要介质。