摘要
慢性胰腺炎是临床常见病,80-90%的慢性胰腺炎病人都有持续性腹部疼痛,其严重的、反复发作的腹部疼痛是病人的主要症状之一。然而,慢性胰腺炎导致疼痛的机制尚不明确,解释其引起疼痛的机制也有很多,包括胰腺本身和胰腺外部原因。关于神经病理性疼痛的研究表明中枢神经系统的疼痛传导通路发生了异常。有研究发现在某些胰腺病变,包括慢性胰腺炎、胰腺癌和胰性脑病,可以导致“神经重塑“和胰腺神经支配发生变化。以上结果提示中枢神经系统可能参与了慢性胰腺炎导致的慢性疼痛。
脊髓背角神经-免疫系统和神经元-神经胶质细胞的相互作用在神经可塑性变化和神经病理性痛中起了十分重要的作用。有研究表明神经-免疫系统相互作用在慢性胰腺炎疼痛中非常重要。以往表明脊髓背角星形胶质细胞在大鼠慢性胰腺炎模型发生了活化,同时抑制星形胶质细胞的活性可以减轻慢性胰腺炎引起的疼痛。我们推测星形胶质细胞在此过程中可能被某些受体活化,然后导致一系列分子表达变化和信号通路的活化,参与了慢性胰腺炎引起的疼痛。但是具体那种分子参与了慢性胰腺炎引起的星形胶质细胞的活化未见报道。Toll样受体(TLRs)在慢性痛病理条件下脊髓背角神经-免疫相互作用和神经元-神经胶质细胞相互作用中起到了十分重要的作用。有学者表明TLRs在中枢神经系统中,特别是神经胶质细胞,有广泛表达。在哺乳动物神经胶质细胞中TLRs的表达已经被确认。在这些受体中,TLR2-4被认为主要介导了神经病理性疼痛。通常来说,在内源性或是外源性信号通路的刺激下,活化的TLRs能够导致神经胶质细胞的活化,调节疼痛相关的炎症因子。有趣的是,TLRs同样有可能在胰腺炎的发生过程中发挥了作用。最近有报道表明腹腔内注射TLR3可以成功的引起慢性胰腺炎样的病理变化。在本实验中,我们假设TLRs,尤其是TLR2-4,参与了慢性胰腺炎病理条件下星形胶质细胞的活化和疼痛的发生。我们的实验结果表明脊髓TLR3参与了大鼠慢性胰腺炎模型的疼痛。慢性胰腺炎病理条件下脊髓的“TLR3-星形胶质细胞- IL-1β/MCP-1”的正反馈通路可能为TLR参与慢性胰腺炎疼痛的机制,或许可以成为临床慢性胰腺炎病人疼痛的缓解的治疗靶点。
目的:
慢性胰腺炎导致的疼痛的机制目前为止尚不明确。以前的很多结果表明星形胶质细胞在疼痛过程中得到了活化。尽管如此,星形胶质细胞在慢性胰腺炎病理过程中是否发生了活化以及其发生活化的分子机制尚不明确。本实验的目的为探讨星形胶质细胞在慢性胰腺炎病理过程中是否发生了活化以及TIRs是否参与星形胶质细胞的活化和慢性胰腺炎引起的疼痛发作。
方法:
1.动物分为慢性胰腺炎实验组,假手术组和正常组。雄性Sprague–Dawley大鼠,体重250-300g,由第四军医大学动物实验中心提供。术前大鼠禁食水12h。麻醉实验动物,常规消毒,沿腹白线做正中切口剖腹,寻找十二指肠并找到胰腺,用微型动脉夹阻断胰胆管肝门部。用4号静脉头皮针经肠壁穿刺入胰胆管,并用丝线固定静脉针,通过恒流静脉输液泵注射含2%TNBS的PBS(PH 7.4),共0.4ml,持续60min,注毕后关闭恒流泵,维持压力50min。对照组注射等体积的生理盐水。除去动脉夹及丝线,逐层关闭腹腔。整个手术操作过程严格执行无菌操作。检测各组血清和胰腺组织细胞因子(TNF-?、IL-1。COX-1和α-SMA)的表达变化。分别于0点,1周和5周对各组大鼠胰腺HE染色后进行病理评分。同时从0点到第5周实时监测各组大鼠的RFs值。利用免疫荧光和RE-PCR检测星形胶质细胞标志物在不同时间点的表达,west blotting检测TLR2-4在不同时间点的表达。免疫荧光双标研究TLR3与星形胶质细胞标志物的共存关系。
2.慢性胰腺炎组给予LAA拮抗星形胶质细胞的活性,利用RT-PCR验证LAA拮抗效果;同时检测RFs值的和星形胶质细胞标志物的变化情况。
3.慢性胰腺炎组给予TLR3反义探针拮抗TLR3的表达,利用west blotting验证TLR3反义探针拮抗效果;同时检测RFs值的和星形胶质细胞标志物的变化情况。
4. RT-PCR检测各组大鼠脊髓细胞因子,如TNF-?, IL-1, IL-6,COX2和MCP-1的变化情况。
结果:
1.在假手术组和正常组,胰腺组织HE染色表现正常。在TNBS注射5周后,实验组大鼠胰腺HE染色表现出明显的胰腺腺泡萎缩,炎细胞浸润,组织纤维化和间质增生。血清淀粉酶、TNF-?和IL-1在实验组显著升高。胰腺组织的TNF-?、IL-1、COX-1和α-SMA显著升高。在此同时,TNBS注射可导致RFs值的从开始到第5周的持续性升高。而假手术组和正常组RFs值比较无统计学意义。TNBS注射组5周时RFs值与刺激正相关。与临床慢性胰腺炎病人症状非常相似。因此我们确认TNBS可导致大鼠慢性胰腺炎以及腹部疼痛敏感度的增高。
2.我们在不同时间点处理各组大鼠观察星形胶质细胞活化标志物GFAP表达的变化情况。免疫荧光和RT-PCR结果显示在假手术组和正常组,GFAP在胸椎背角的表达很低。在实验组,有趣的是,脊髓GFAP的表达从TNBS注射后第1周到第5周显著升高,第5周达到高峰,其表达的变化与疼痛的变化成正相关。因此我们假设GFAP可能介导慢性胰腺炎导致的疼痛。
3.我们在不同时间点处理各组大鼠观察TLR2-4蛋白表达的变化情况。Western blotting结果显示在假手术组和正常组,TLR2-4在胸椎背角的表达很低。在实验组,有趣的是,脊髓TLR3的表达从TNBS注射后第一周到第五周显著升高,第5周达到高峰,其表达的变化与疼痛的变化成正相关,这与以往我们的研究表明脊髓星形胶质细胞的表达变化非常一致。因此我们假设TLR3可能在星形胶质细胞上表达,同时介导慢性胰腺炎导致的星形胶质细胞的活化。免疫双标显示星形胶质细胞特异性标志物GFAP与TLR3的共表达,初步证实了我们的假设。
4.接下来我们鞘内注射GFAP的活性抑制剂LAA。RT-PCR证实了其拮抗效果。鞘内注射LAA在一定程度上可以缓解慢性胰腺炎引起的疼痛,这种效应为计量依赖性。在假手术组或正常组,鞘内注射LAA对RFs无影响。这些结果提示GFAP有可能参与到慢性胰腺炎导致的疼痛。接下来我们鞘内注射TLR3的反义探针拮抗TLR3的功能,western blotting证实了其拮抗效果。鞘内注射TLR3的反义探针在一定程度上可以缓解慢性胰腺炎引起的疼痛,这种效应为计量依赖性。在假手术组或正常组,鞘内注射TLR3反义探针对RFs无影响。这些结果暗示了TLR3有可能参与到慢性胰腺炎导致的疼痛。考虑到TLR3在脊髓星形胶质细胞的高表达,接下来我们探讨了TLR3反义探针对于慢性胰腺炎介导的星形胶质细胞活化的作用。我们实验结果发现TLR3反义探针显著抑制了脊髓星形胶质细胞的活化。然后我们又检测了脊髓炎症因子的表达情况,脊髓IL-1?,TNF-?,IL-6和MCP-1在慢性胰腺炎组表达明显升高,而只有TNF-?和IL-6的mRNA表达不受TLR3反义探针的影响。有趣的是,脊髓COX2在正常组和胰腺炎组表达无显著差异,TLR3反义探针也不影响其在脊髓的表达。
结论:
以上结果表明脊髓星形胶质细胞和其表达的TLR3参与了大鼠慢性胰腺炎模型的疼痛。慢性胰腺炎病理条件下脊髓的“TLR3-星形胶质细胞- IL-1β/MCP-1”的正反馈通路可能为TLR参与慢性胰腺炎疼痛的机制,或许可以成为临床慢性胰腺炎病人疼痛的缓解的治疗靶点。
Chronic pancreatitis (CP) is a severe inflammatory and painful disease of the exocrine pancreas. Constant, recurrent and serious abdominal pain is one of the most common symptoms in CP, present in 80–90% of the patients. However,the pain mechanisms in CP are incompletely understood and probably are multifactorial,including pancreatic and extrapancreatic causes. Experimental human pain studies show that pain processing in the central nervous system (CNS) is abnormal in neuropathic pain disorders. A recent study shows that in the patients of CP and pancreatic cancer, pancreatic neuropathy could bring "neural remodeling" and alter pancreatic innervation. These results highly suggest that neuroplastic changes in the CNS are probably important contributors to the CP-induced chronic pain. Neuron-immune interactions and neuron-glial crosstalk in the spinal dorsal horn play a pivotal role in neuroplastic changes and neuropathic pain. It has been pointed out that neuroimmune interactions are significant in pain generation in CP. Our recent study observed that astrocytes are activated in the thoracic spinal cord in a rat model of CP induced by intrapancreatic infusion of trinitrobenzene sulfonic acid (TNBS). While inhibiting astrocytic activation could attenuate pain of CP. We can estimate that, like in that of nerve injury-induced neuropathic pain states, astrocytes might be activated through some receptors on it, and then produce signaling molecules that can feed back to enhance neuronal activity, contributing to pain facilitation. However,it was still unclear,in CP conditions:which receptor(s) mediated astrocytic activation. Toll-like receptors (TLRs) play a key role in neuron-immune interactions and neuron-glial crosstalk in the spinal dorsal horn in chronic pain conditions . It has been demonstrated that TLRs are widely expressed in the CNS, particularly by glial cells. In mammals, 12 members of the TLR family have been identified so far. Among them, TLR2-4 have been clarified to be major mediators in neuropathic pain. In general,in response to stimulation by endogenous and exogenous signals,activation of TLRs could induce glial activation in which multiple TLRs could trigger and tailor innate immune responses of glia by altering production of pain-associated pro-inflammatory cytokines/chemokines. However, there is still no report on the contribution of TLRs in CP related pain. Interestingly, TLRs have been implicated in the process of pancreatitis. A recent study even showed that intraperitoneal injection of TLR 3 activator could successfully induce CP-like pathological changes. In the present study, we hypothesized that TLRs,especially TLR2-4,contributed to astrocytic activation and pain behavior in the process of CP-induced pain. To test our hypothesis, we first observed the expression changes of TLR2-4 following TNBS-induced CP. We found that TLR3, but not TLR2 or 4 was increased in the thoracic spinal dorsal horn in the process of CP. Then we detected the cellular localization of TLR3 with double immunostaining and
observed these TLR3 were highly expressed on spinal astrocytes. We further used a kind of TLR3 antisense oligodeoxynucleotide (ASO) to decrease the expression of TLR3 and observed the behavioral and biochemical changes in the spinal cord.
OBJECTIVE:
Mechanisms underlying pain in chronic pancreatitis (CP) are incompletely understood. Our previous data showed that astrocytes were actively involved. However, the molecular basis for astrocytic activation was unclear. The objective of this study is to investigate whether toll-like receptors (TLRs) contributed to astrocytic activation and pain behavior in CP-induced pain.
METHODS:
1. Induction of pancreatitis
Male Sprague–Dawley rats were used in the study. Animals were given free access to drinking water and standard food pellets until 12 h prior to induction of pancreatitis, at which point food was withdrawn. Briefly, the common bile duct was closed temporarily near the liver with a small vascular clamp. A blunt 28 gauge needle with PE 10 tubing attached was inserted into the duodenum and was guided through the papilla into the duct and was secured with suture. TNBS solution (0.5 ml, 2%) in 10% ethanol in phosphate buffered saline (PBS,pH 7.4) was infused into the pancreatic duct over a period of 2–5 min at a pressure of 50 mmHg. After 30 min exposure to TNBS, needle and tubing were removed. The hole in the duodenum was sutured and the vascular clamp was removed restoring the bile flow. All the procedures in the sham group were same to that of the TNBS group, except that the same volume of saline instead of TNBS was infused into the duct.
2. Administration of drugs
For single intrathecal injection, we performed spinal cord puncture under sevo uorane anesthesia with a 27 gauge needle between the T9 and T12 level to deliver the reagents (20 l) to the cerebrospinal ?uid. Immediately after the needle entry into subarachnoid space, a brisk tail ?ick could be observed. Intrathecal injection of LAA (Sigma, St. Louis, MO, USA; 50, 100, and 150 nmol in 10 l saline) or saline (10 l) was performed on the 34th d (1 d before 5 weeks post-operation) after pancreatitis induction. TLR3 ASO (5’- AACAATTGCTTCAAGTCC TCAAGTCC-3’) and TLR3 mismatch ODN (5’-ACTTCAACAGTAGACTACTAGACTAC-3’) were synthesized by Sangon Biotechnology Co.. For intrathecal infusion, laminectomy was performed at the level of the thoracic vertebrae, under pentobarbital anesthesia (45 mg kg-1 ,i.p.). A polyethylene (PE10) catheter (I.D. 0.28 mm and O.D. 0.61 mm) was passed caudally from the T9 to the T12 level of the spinal cord,and 2 cm of the free ending was left exposed in the upper thoracic region. The catheter was connected to an osmotic pump was filled with AS-ODN (0.01, 0.1or 1 nmol/μL) or MM-ODN (1 nmol/lL) in normal saline.
All the rats were divided into three groups: TNBS group (n = 54); sham group (n =42) and na?ve controls (n = 6). In order to study the time course of TLRs change,we sacrificed every 6 rats in TNBS group at 1,2,3,4 and 5 w after TNBS infusion. At 4 w after surgery,other rats in TNBS and sham groupwere each further divided for drug injection:TNBS-ASO (or sham-ASO) group: TLR3 ASO was intrathecally infused on pancreatitic (0.01,0.1 or 1 nmol/μL) or sham operated rats (1 nmol/μL); TNBS-MO (or sham-MO) group: TLR3 MO was injected on pancreatitic or sham operated rats. At 5 w after surgery,all the rats including na?ve rats and sham rats without intrathecal injection were sacrificed for further experiments.
3. Behavioral tests
Mechanical allodynia was measured with von Frey filaments. Briefly, the belly was shaved prior to test and areas designated for stimulation were marked. Rats were placed in a plastic cage with a mesh floor and were given 30 min for adaptation before testing. The von Frey filaments were applied from underneath through the mesh floor,in ascending order to the abdominal area at different points on the surface. A single trial consisted of 10 applications each for 1–2 s with a 15 s interval between applications allow the animal to cease any response and return to a relatively inactive position. A positive response consisted of the rat raising its belly (withdrawal response). The data were expressed as a percentage of the number of positive responses with each filament for each rat. In the time-course study, behavioral tests were performed before and once weekly for up to 5 w after induction of pancreatitis with a single force of von Frey filament (40.7 mN). In the strength-response study, rats were tested 5 w after pancreatitis induction with a series of von Frey filaments (2.29, 2.75, 6.76, 16.6, 40.7, 69.2 and 120 mN).
4. Pancreatic histology
Rats were deeply anesthetized with sodium pentobarbital (60 mg/kg,i.p.) and the pancreas was obtained and then fixed in 4% paraformaldehyde in phosphate buffered (PB,pH 7.4) at 4℃overnight. Pancreatic tissue was then transferred to xylene washes twice and was placed in cassettes and embedded in paraffin. Paraffin blocks were cut in 5-μm sections and stained with hematoxylin and eosin. Histological sections were analyzed by a pathologist in a double blinded manner. The severity of CP was morphologically assessed by semiquantitative scores according to previous reports : graded glandular atrophy (0–3),intralobular,interlobular and periductal fibrosis (0–3) and inflammatory cells infiltration (0–3).
5. reverse transcription polymerase chain reaction for Pancreatic tissues
RT-PCR was performed to analyze the mRNA expression levels of TNF-α, IL-1β, COX-2 and IL-10 in the pancreatic tissue. RNA was extracted from pancreatic tissue using Trizol reagent. One microgram of RNA was reverse transcribed to cDNA. Amplification was performed with the following cycles: 95°C for 30 s,followed by 40 cycles of denaturing at 95°C for 5 s and annealing at 60°C for 20 s. All of the reactions were performed in triplicate.
6. Biochemical Assays
Serum TNF-α, IL-1βand IL-10 were determined by ELISA. The serum TNF-α, IL-1βand IL-10 levels were evaluated using ELISA kits.
7. Western blotting
All animals were rapidly sacrificed and the thoracic (T) 10 spinal dorsal horn was rapidly harvested. Spinal cord was then homogenized with a hand-held pestle in SDS sample buffer (10 ml/mg tissue), which contained a cocktail of proteinase and phosphatase inhibitors. The electrophoresis samples were heated at 100°C for 5 min and loaded onto 10% SDS-polyacrylamide gels with standard Laemmli solutions. The proteins were electroblotted onto a polyvinylidene difluoride membrane. The membranes were placed in a blocking solution containing Tris-buffered saline with 0.02 % Tween (TBS-T) and 5% non-fat dry milk,for 1 h,and incubated overnight under gentle agitation with primary antibodies: rabbit anti-TLR2,rabbit anti-TLR3,rabbit anti-TLR4 and rabbit anti-?-actin. Bound primary antibodies were detected with the anti-rabbit horseradish peroxidase (HRP)-conjugated secondary antibody. Between each step,the immunoblots were rinsed with TBS-T. All reactions were detected by the enhanced chemiluminescence (ECL) detection method. The densities of protein blots were analyzed by using Labworks Software. The densities of target proteins and b-actin immunoreactive bands were quantified with background subtraction. The same size of square was drawn around each band to measure the density and the background near that band was subtracted. Target protein levels were normalized against ?-actin levels and expressed as relative fold changes compared to the na?ve control or to the sham-MO group.
8. Real-time reverse transcription polymerase chain reaction (RT-PCR)
Rats were deeply anesthetized with sodium pentobarbital (60 mg/kg,i.p.) and then thoracic (T) 10 spinal dorsal horn was rapidly harvested and RNA was extracted with Trizol. Complementary DNA (cDNA) was synthesized with oligo (dT)12-18 using SuperscriptTMⅢReverse Transcriptase for RT-PCR. The primers used in the present study were presented in Table. 1. Equal amounts of RNA (1 ?g) were used to prepare cDNA using the SYBR? Premix Ex Taq? and analyzed by real-time PCR in a detection system. The amplification protocol was: 3 min at 95°C, followed by 45 cycles of 10 s at 95℃for denaturation and 45 s at 60℃for annealing and extension. All experiments were repeated twice and, in each experiment, PCR reactions were done in triplicate. Target cDNA quantities were estimated from the threshold amplification cycle number (Ct) using Sequence Detection System software. GAPDH was served as an endogenous internal standard control for variations in RT-PCR efficiency.
9. Immunofluorescent double labeling
At 5 w after TNBS infusion,rats were perfused through the ascending aorta with 100 ml of normal saline followed by 500 ml of 0.1 M phosphate buffer containing 4% paraformaldehyde and 2% picric acid,under deep anesthesia with sodium pentobarbital (60 mg/kg,i.p.). After the perfusion,the spinal segments T10 was removed and postfixed in the same fixative for 2-4 h and then cryoprotected for 24 h at 4°C in 0.1 M PB containing 30% sucrose. Transverse spinal sections (30μm thickness) were cut in a cryostat, collected in 0.01 M phosphate-buffered saline (PBS, pH 7.3) and were then processed for immunofluorescent staining. Sections were rinsed in 0.01 M PBS (pH 7.3) for three times (10 min each),blocked with 2% goat serum in 0.01 M PBS containing 0.3% Triton X-100 for 1 h at the room temperature (RT) and then used for immunofluorescent staining. The sections were incubated overnight at 4°C with the primary antibodies: mouse anti-GFAP. The sections were then washed for three times in 0.01 M PBS (10 min each) and then incubated for 2 h at RT with the corresponding secondary antibody: FITC-conjugated horse anti-mouse IgG and Alexa 594-conjugated donkey anti-rabbit IgG. Images were obtained using a confocal laser microscope and Digital images were captured with Fluoview 1000.
RESULTS:
1. TNBS infusion induced CP and mechanical allodynia
In the na ve and sham-operated rats, the pancreas presented a normal appearance. While on 5 w after TNBS infusion, the pancreas showed significant acinar atrophy, inflammatory infiltration, and periductular and intralobular fibrosis, stromal proliferation. CP-induced persistent mechanical allodynia is characterized by increase of abdomen response frequencies shown by increase of abdomen response frequencies (RFs). We observed in the time course study that rats with CP showed a persistent mechanical hypersensitivity in the abdomen that was evident 1 w after TNBS infusion and persisted up to 5 w. There was no significant difference between sham and naive group at every corresponding time point. Then, we examined the sensitivity of rats with different treatments via von-Frey filaments of various strengths from 2.29 to 120 mN on 5 w post CP induction. We observed that RFs of rats 5 w after TNBS infusion were significantly higher at all filaments tested compared to either sham or to naive rats. Thus we confirmed that intrapancreatic infusion of TNBS produced CP and a significant increase in sensitivity to mechanical probes in the abdomen.
2. Astrocytic activation in the thoracic spinal cord in pain of CP
GFAP staining wassignificantly increased in the spinal cord 5 weeks after CP induction. This enhanced staining appeared in all areas of the spinal grey matter, with the most prominent increase in superficial dorsal horn laminae I–III.
3. Inhibition of astrocytic activation attenuated TNBS-induced mechanical allodynia
The astrocytic specific toxin decreased spinal GFAP expression in the sham- operated rats (P 0.05 vs. sham-saline group and pancreatic rats. Meanwhile, we analyzed behavioral changes of the rats following different treatments. Treatment with LAA significantly, even though not completely, attenuated the allodynia.
4. CP significantly up-regulated TLR3 expression in the thoracic spinal dorsal horn
We then sacrificed the rats at different time points to observe the expression changes of TLRs expressions. Western blot indicated that in na?ve or sham operated rats, TLRs expressions in the thoracic spinal dorsal horn were at very low levels. After intrapancreatic infusion of TNBS, TLR 2 and TLR 4 were still at a very low level, compared with that of na?ve and sham group. Very interestingly, TLR3 was significantly increased in the spinal cord, since 1 w after CP induction. TLR3 was increased gradually and maintained at a very high level up to 5 w, which correlated with the changing course of mechanical allodynia. In our previous study, we observed a similar changing course of spinal astrocytic activation after CP induction, so we made a hypothesis that TLR3 could be expressed on spinal astrocytes and mediate CP-induced astrocytic activation. We thus observed the cellular localization of TLR3 in the thoracic spinal dorsal horn 5 w after CP induction. By double immunostaining with antibodies against astrocytic specific marker GFAP and TLR3, we observed that TLR3 was highly expressed on spinal astrocytes.
5. Intrathecal infusion of TLR3 ASO significantly attenuated CP-induced mechanical allodynia
In order to testify our hypothesis that TLR3 contributed to CP-induced neuropathic pain, we used a kind of TLR3 ASO to knockdown the expression of TLR3 and observe the behavioral consequences and cellular and molecular changes. Infusion with TLR3 ASO (0.1 nmol/μl, from 4th w to 5th w following TNBS infusion) significantly, even though not completely, attenuated the allodynia. However, TLR3 ASO did not influence RFs of sham operated rats. We further observed that TNBS-induced allodynia was significantly attenuated by TLR3 ASO, in a dose-dependent manner. Western blot confirmed that intrathecal infusion of TLR3 ASO, but not TLR3 MO, significantly blocked CP-induced TLR3 up-regulation. These results suggested that TLR3 might contribute to CP-induced mechanical allodynia. Considering the high expression of TLR3 on spinal astrocytes after CP induction, we thus investigated the role of TLR3 ASO on CP-induced astrocytic activation.
6. TLR3 ASO significantly reversed CP-induced astrocytic activation,as well as cytokines expressions
Very interestingly, we observed that CP-induced astrocytic activation in the thoracic spinal dorsal horn was remarkably suppressed by TLR3 ASO. GFAP expression in the TNBS-MO group was significantly higher than that of sham group. However, GFAP levels in the TLR3 ASO groups were much lower than that of the TNBS-MO group, although still higher than that of the sham groups. We then further observed the cytokines expression in the rat thoracic spinal dorsal horn following different treatments. A significant up-regulation of cytokines was observed after CP-induced chronic pain. In the TNBS-MO group,we observed significant increases of IL-1?,TNF-?,IL-6 and MCP-1 compared with that of sham groups. Intrathecal infusion of TLR3 ASO could significantly attenuated CP-induced up-regulation of IL-1? and monocyte chemotactic protein-1 (MCP-1), in a dose dependent manner. However, TNF-? or IL-6 was not significantly influenced by TLR3 ASO. Besides, different from other cytokines observed, COX2 in the thoracic spinal dorsal was not increased in CP conditions,either influenced by intrathecal infusion of TLR3 ASO.
CONCLUSIONS:
Our results provide evidence for the involvement of spinal TLR3 in CP-induced chronic pain. And we present a probable“TLR3 - astrocytes - IL-1β/MCP-1”pathway as a positive feedback loop in the spinal dorsal horn in CP conditions,which could be new targets for treating severe and persistent pain in CP patients.
引文
1.王兴鹏,龚自华.慢性胰腺炎的治疗进展.中华消化杂志,2001,21(1):36-38
2.王兴鹏,龚自华.慢性胰腺炎的实验室诊断.胃肠病学,2001,6(3):176-177
3. Su SB,Motoo Y,Xie MJ,Miyazono K,Sawabu N. Expression of transforming growth factor-beta in spontaneous chronic pancreatitis in the WBN/Kob rat. Dig Dis Sci. 2000 Jan;45(1):151-9.
4. Su SB,Motoo Y,Xie MJ,Sakai J,Taga H,Sawabu N. Expression of pancreatitis-associated protein (PAP) in rat spontaneous chronic pancreatitis:effect of herbal medicine Saiko-keishi-to (TJ-10). Pancreas. 1999 Oct;19(3):239-47.
5. Su SB,Motoo Y,Xie MJ,Taga H,Sawabu N. Antifibrotic effect of the herbal medicine Saiko-keishi-to (TJ-10) on chronic pancreatitis in the WBN/Kob rat. Pancreas. 2001 Jan;22(1):8-17.
6. Motoo Y,Su SB,Xie MJ,Taga H,Sawabu N. Effect of herbal medicine Saiko-keishi-to (TJ-10) on rat spontaneous chronic pancreatitis:comparison with other herbal medicines. Int J Pancreatol. 2000 Apr;27(2):123-9.
7. Tsukamoto H,Towner SJ,Yu GSM,et al. Potentiation of ethanol induced pancreatic injury by dietary fat induction of chronic pancreatitis by alcohol in rats.Am J Pathol,1988;131(2):246-57.
8.刘庆彬,白静丽.慢性胰腺炎动物模型研究现状.中国老年学杂志. 2009,29(9):173-176.
9. Neuschwander-Tetri BA,Burton FR,Presti ME,Britton RS,Janney CG,Garvin PR,Brunt EM,Galvin NJ,Poulos JE. Repetitive self-limited acutepancreatitis induces pancreatic fibrogenesis in the mouse. Dig Dis Sci. 2000 Apr;45(4):665-74.
10. van Westerloo DJ,Florquin S,de Boer AM,Daalhuisen J,de Vos AF,Bruno MJ,van der Poll T. Therapeutic effects of troglitazone in experimental chronic pancreatitis in mice. Am J Pathol. 2005 Mar;166(3):721-8
11.万远太,王天才,赵秋,等.蛙皮素致大鼠胰腺慢性纤维化模型的建立.胰腺病学,2005;9(3):158-61.
12. Merkord J,Weber H,Sparmann G,Jonas L,Hennighausen G. The course of pancreatic fibrosis induced by dibutyltin dichloride (DBTC). Ann N Y Acad Sci. 1999 Jun 30;880:231-7.
13. Merkord J,Weber H,Jonas L,Nizze H,Hennighausen G. The influence of ethanol on long-term effects of dibutyltin dichloride (DBTC) in pancreas and liver of rats. Hum Exp Toxicol. 1998 Mar;17(3):144-50.
14. Sparmann G,Behrend S,Merkord J,Kleine HD,Graser E,Ritter T,Liebe S,Emmrich J. Cytokine mRNA levels and lymphocyte infiltration in pancreatic tissue during experimental chronic pancreatitis induced by dibutyltin dichloride. Dig Dis Sci. 2001 Aug;46(8):1647-56.
15.何新红,陆建平,廖专,等.二丁基二氯化物尾静脉注射建立大鼠慢性胰腺炎模型.胰腺病学,2007;7(1):17-20.
16. Vera-Portocarrero LP , Lu Y , Westlund KN. Nociception in persistent pancreatitis in rats : effects of morphine and neuropeptide alterations. Anesthesiology. 2003 Feb;98(2):474-84.
17. Kataoka K,Sasaki T,Yorizumi H,Sakagami J,Kashima K. Pathophysiologic studies of experimental chronic pancreatitis in rats induced by injection of zein-oleic acid-linoleic acid solution into the pancreatic duct. Pancreas. 1998 Apr;16(3):289-99.
18. Haber PS,Keogh GW,Apte MV,Moran CS,Stewart NL,Crawford DH,Pirola RC,McCaughan GW,Ramm GA,Wilson JS. Activation of pancreatic stellate cells in human and experimental pancreatic fibrosis. Am J Pathol. 1999 Oct;155(4):1087-95.
19.王兴鹏,龚自华,吴恺,等.慢性胰腺炎动物模型的建立.胰腺病学.2001,1(1):52-53
20. Andren-Sandberg A,Hoem D,Gislason H. Pain management in chronic pancreatitis. Eur J Gastroenterol Hepatol 2002;14:957–70.
21. Friess H,Shrikhande S,Shrikhande M,Martignoni M,Kulli C,Zimmermann A,Kappeler A,Ramesh H,Büchler M. Neural alterations in surgical stage chronic pancreatitis are independent of the underlying aetiology. Gut 2002;50:682-686
22. Di Sebastiano P,Fink T,Weihe E,Friess H,Innocenti P,Beger HG,Büchler MW. Immune cell infiltration and growth-associated protein 43 expression correlate with pain in chronic pancreatitis. Gastroenterology 1997;112: 1648-1655
23. Elitt CM,McIlwrath SL,Lawson JJ,Malin SA,Molliver DC,Cornuet PK,Koerber HR,Davis BM,Albers KM. Artemin overexpression in skin enhances expression of TRPV1 and TRPA1 in cutaneous sensory neurons and leads to behavioral sensitivity to heat and cold. J Neurosci 2006; 26: 8578-8587
24. Malin SA,Molliver DC,Koerber HR,Cornuet P,Frye R,Albers KM,Davis BM. Glial cell line-derived neurotrophic factor fami ly members sensitize nociceptors in vitro and produce thermal hyperalgesia in vivo. J Neurosci 2006;26:8588-8599
25.廖兴志,熊源长.神经性疼痛机制在慢性胰腺炎中的作用.世界华人消化杂志. 2009;17(31):3229-3232
26. Joos GF,Pauwels RA. Pro-inflammatory effects of substance P:new perspectives for the treatment of airway diseases? Trends Pharmacol Sci. 2000 Apr;21(4):131-3.
27. Bhatia M,Brady M,Shokuhi S,Christmas S,Neoptolemos JP,Slavin J. Inflammatory mediators in acute pancreatitis. J Pathol. 2000 Feb;190(2):117-25
28.徐飞,汤雨,千年松.坐骨神经分支选择性损伤后降钙素相关肽在大鼠中央杏仁核中的表达变化.神经解剖学杂志,2011:27(1):14-18
29. Friess H,Zhu ZW,di Mola FF,Kulli C,Graber HU,Andren-Sandberg A,Zimmermann A,Korc M,Reinshagen M,Büchler MW. Nerve growth factor and its high-affinity receptor in chronic pancreatitis. Ann Surg. 1999 Nov;
30. Cho HJ,Kim SY,Park MJ,Kim DS,Kim JK,Chu MY. Expression of mRNA for brain-derived neurotrophic factor in the dorsal root ganglion following peripheral inflammation. Brain Res. 1997 Feb 28;749(2):358-62.
31. Liddle RA. The role of Transient Receptor Potential Vanilloid 1 (TRPV1) channels in pancreatitis. Biochim Biophys Acta 2007;1772:869-878
32. Immke DC,Gavva NR. The TRPV1 receptor and nociception. Semin Cell Dev Biol 2006;17:582-591
33. Hartel M,di Mola FF,Selvaggi F,Mascetta G,Wente MN,Fel ix K,Giese NA,Hinz U,Di Sebastiano P,Büchler MW,Friess H. Vanilloids in pancreatic cancer:potential for chemotherapy and pain management. Gut 2006;55:519-528
34. DeRosier LC,Huang ZQ,Sellers JC,Buchsbaum DJ,Vickers SM. Treatment with gemcitabine and TRA-8 anti-death receptor-5 mAb reduces pancreatic adenocarcinoma cell viability in vitro and growth in vivo. J Gastrointest Surg.2006 Nov;10(9):1291-300;
35. Loeser JD,Treede RD. The Kyoto protocol of IASP basic pain terminology. Pain 2008;137:473–7.
36. Liu XG,Sandkuhler J . Activat ion of spinal N-methyl-D-aspartate or neurokinin receptors induces long-term potentiation of spinal C-fibre-evoked potentials. Neuroscience 1998;86:1209-1216
37. van Esch AA,Wilder-Smith OH,Jansen JB,van Goor H,Drenth JP. Pharmacological management of pain in chronic pancreatitis. Dig Liver Dis 2006;38:518-526
38. Giamberardino MA. Recent and forgotten aspects of visceral pain. Eur J Pain 1999;3:77-92
39. Vera-Portocarrero LP,Xie JY,Kowal J,Ossipov MH,King T,Porreca F. Descending facilitation from the rostral ventromedial medulla maintains visceral pain in rats with experimental pancreatitis. Gastroenterology 2006; 130:2155-2164
40. Drewes AM,Reddy H,Pedersen J,Funch-Jensen P,Gregersen H,Arendt-Nielsen L. Multimodal pain stimulations in patients with grade B oesophagitis. Gut 2006; 55: 926-932
41. Dimcevski G,Sami SA,Funch-Jensen P,Le Pera D,Valeriani M,Arendt-Nielsen L,Drewes AM. Pain in chronic pancreatitis: the role of reorganization in the central nervous system. Gastroenterology 2007; 132: 1546-1556
42. Cotter DR,Pariante CM,Everall IP. Glial cell abnormalities in major psychiatric disorders:the evidence and implications. Brain Res Bull. 2001 Jul 15;55(5):585-95.
43. Rouach N,Glowinski J,Giaume C. Activity-dependent neuronal control ofgap-junctional communication in astrocytes. J Cell Biol. 2000 Jun 26;149(7):1513-26
44. Dreyer EB,Leifer D,Heng JE,McConnell JE,Gorla M,Levin LA, Barnstable CJ,Lipton SA. An astrocytic binding site for neuronal Thy-1 and its effect on neurite outgrowth. Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):11195-9.
45. Garrison CJ,Dougherty PM,Kajander KC,Carlton SM.Staining of glial fibrillary acidic protein (GFAP) in lumbar spinal cord increases following a sciatic nerve constriction injury. Brain Res. 1991 Nov 22;565(1):1-7.
46. Raghavendra V,Tanga FY,DeLeo JA. Complete Freunds adjuvant-induced peripheral inflammation evokes glial activation and proinflammatory cytokine expression in the CNS. Eur J Neurosci. 2004 Jul;20(2):467-73.
47. Sweitzer SM,Colburn RW,Rutkowski M,DeLeo JA. Acute peripheral inflammation induces moderate glial activation and spinal IL-1beta expression that correlates with pain behavior in the rat. Brain Res. 1999 May 22;829(1-2):209-21.
48. Hald A,Nedergaard S,Hansen RR,Ding M,Heegaard AM. Differential activation of spinal cord glial cells in murine models of neuropathic and cancer pain. Eur J Pain. 2009 Feb;13(2):138-45.
49. Zhang J,De Koninck Y. Spatial and temporal relationship between monocyte chemoattractant protein-1 expression and spinal glial activation following peripheral nerve injury. J Neurochem. 2006 May;97(3):772-83.
50. O'Callaghan JP,Sriram K. Glial fibrillary acidic protein and related glial proteins as biomarkers of neurotoxicity. Expert Opin Drug Saf. 2005 May;4(3):433-42
51. Ahmed Z,Lewis CA,Faber DS. Glutamate stimulates release of Ca2+ frominternal stores in astroglia. Brain Res. 1990 May 14 ; 516(1) : 165-9. Cholewinski AJ,Hanley MR,Wilkin GP. A phosphoinositide-linked peptide response in astrocytes:evidence for regional heterogeneity. Neurochem Res. 1988 Apr;13(4):389-94.
52. Gallagher CJ,Salter MW. Differential properties of astrocyte calcium waves mediated by P2Y1 and P2Y2 receptors. J Neurosci. 2003 Jul 30;23(17):6728-39.
53. Gu JG. P2X receptor-mediated modulation of sensory transmission to the spinal cord dorsal horn. Neuroscientist. 2003 Oct;9(5):370-8.
54. Haas CA,Reddington M,Kreutzberg GW. Calcitonin Gene-related Peptide Stimulates the Induction of c-fos Gene Expression in Rat Astrocyte Cultures. Eur J Neurosci. 1991 Jul;3(7):708-712.
55. Lazar P,Reddington M,Streit W,Raivich G,Kreutzberg GW. The action of calcitonin gene-related peptide on astrocyte morphology and cyclic AMP accumulation in astrocyte cultures from neonatal rat brain. Neurosci Lett. 1991 Sep 2;130(1):99-102
56. Neary JT,Baker L,Jorgensen SL,Norenberg MD. Extracellular ATP induces stellation and increases glial fibrillary acidic protein content and DNA synthesis in primary astrocyte cultures. Acta Neuropathol. 1994;87(1):8-13.
57. Noble LJ,Hall JJ,Chen S,Chan PH. Morphologic changes in cultured astrocytes after exposure to glutamate. J Neurotrauma. 1992 Fall;9(3):255-67.
58. Garrison CJ,Dougherty PM,Carlton SM. GFAP expression in lumbar spinal cord of naive and neuropathic rats treated with MK-801. Exp Neurol. 1994 Oct;129(2):237-43.
59. Steward O,Torre ER,Tomasulo R,Lothman E. Neuronal activity up-regulates astroglial gene expression. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6819-23.
60. Del Bigio MR,Fedoroff S,Qualtiere LF. Morphology of astroglia in colony cultures following transient exposure to potassium ion,hypo?smolarity and vasopressin. J Neurocytol. 1992 Jan;21(1):7-18.
61. Del Bigio MR,Fedoroff S,Qualtiere LF. Morphology of astroglia in colony cultures following transient exposure to potassium ion, hypo?smolarity and vasopressin. J Neurocytol. 1992 Jan;21(1):7-18.
62. Neprasova H,Anderova M,Petrik D,Vargova L,Kubinova S,Chvatal A, Sykova E. High extracellular K(+) evokes changes in voltage-dependent K(+) and Na (+) currents and volume regulation in astrocytes. Pflugers Arch. 2007 Mar;453(6):839-49.
63. Guo W,Zingg JM,Meydani M,Azzi A. Alpha-Tocopherol counteracts ritonavir-induced proinflammatory cytokines expression in differentiated THP-1 cells. Biofactors. 2007;31(3-4):171-9.
64. Hald A,Lotharius J. Oxidative stress and inflammation in Parkinson's disease:is there a causal link? Exp Neurol. 2005 Jun;193(2):279-90.
65. Honore P,Rogers SD,Schwei MJ,Salak-Johnson JL,Luger NM,Sabino MC,Clohisy DR,Mantyh PW. Murine models of inflammatory, neuropathic and cancer pain each generates a unique set of neurochemical changes in the spinal cord and sensory neurons. Neuroscience. 2000;98(3):585-98.
66. Murray M,Wang SD,Goldberger ME,Levitt P. Modification of astrocytes in the spinal cord following dorsal root or peripheral nerve lesions. Exp Neurol. 1990 Dec;110(3):248-57.
67. Kozlova EN. Differentiation and migration of astrocytes in the spinal cord following dorsal root injury in the adult rat. Eur J Neurosci. 2003 Feb;17(4):782-90.
68. Ji RR,Kawasaki Y,Zhuang ZY,Wen YR,Decosterd I. Possible role of spinal astrocytes in maintaining chronic pain sensitization: review of current evidence with focus on bFGF/JNK pathway. Neuron Glia Biol. 2006 Nov;2(4):259-269.
69. Krishna M,Narang H. The complexity of mitogen-activated protein kinases (MAPKs) made simple. Cell Mol Life Sci. 2008 Nov;65(22):3525-44.
70. Ma W,Quirion R. Partial sciatic nerve ligation induces increase in the phosphorylation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) in astrocytes in the lumbar spinal dorsal horn and the gracile nucleus. Pain. 2002 Sep;99(1-2):175-84.
71. Falsig J,P?rzgen P,Lotharius J,Leist M. Specific modulation of astrocyte inflammation by inhibition of mixed lineage kinases with CEP-1347. J Immunol. 2004 Aug 15;173(4):2762-70.
72. Zhuang ZY,Wen YR,Zhang DR,Borsello T,Bonny C,Strichartz GR,Decosterd I,Ji RR. A peptide c-Jun N-terminal kinase (JNK) inhibitor blocks mechanical allodynia after spinal nerve ligation: respective roles of JNK activation in primary sensory neurons and spinal astrocytes for neuropathic pain development and maintenance. J Neurosci. 2006 Mar 29;26(13):3551-60.
73. Gao YJ,Ji RR. Activation of JNK pathway in persistent pain. Neurosci Lett. 2008 Jun 6;437(3):180-3.
74. Bogoyevitch MA,Kobe B. Uses for JNK:the many and varied substrates of the c-Jun N-terminal kinases. Microbiol Mol Biol Rev. 2006 Dec;70(4):1061-95.
75. Gühring H,G?rig M,Ates M,Coste O,Zeilhofer HU,Pahl A,Rehse K,Brune K. Suppressed injury-induced rise in spinal prostaglandin E2 production and reduced early thermal hyperalgesia in iNOS-deficient mice. J Neurosci. 2000 Sep 1;20(17):6714-20.
76. Hashizume H,DeLeo JA,Colburn RW,Weinstein JN. Spinal glial activation and cytokine expression after lumbar root injury in the rat. Spine (Phila Pa 1976). 2000 May 15;25(10):1206-17.
77. Winkelstein BA,Rutkowski MD,Sweitzer SM,Pahl JL,DeLeo JA. Nerve injury proximal or distal to the DRG induces similar spinal glial activation and selective cytokine expression but differential behavioral responses to pharmacologic treatment. J Comp Neurol. 2001 Oct 15;439(2):127-39.
78. Liu GJ,Kalous A,Werry EL,Bennett MR. Purine release from spinal cord microglia after elevation of calcium by glutamate. Mol Pharmacol. 2006 Sep;70(3):851-9.
79. Sachs D,Cunha FQ,Poole S,Ferreira SH. Tumour necrosis factor-alpha, interleukin-1beta and interleukin-8 induce persistent mechanical nociceptor hypersensitivity. Pain. 2002 Mar;96(1-2):89-97
80. Trebino CE,Stock JL,Gibbons CP,Naiman BM,Wachtmann TS,Umland JP,Pandher K,Lapointe JM,Saha S,Roach ML,Carter D,Thomas NA,Durtschi BA,McNeish JD,Hambor JE,Jakobsson PJ,Carty TJ,Perez JR, Audoly LP. Impaired inflammatory and pain responses in mice lacking an inducible prostaglandin E synthase. Proc Natl Acad Sci U S A. 2003 Jul 22;100(15):9044-9.
81. Benveniste EN,Sparacio SM,Norris JG,Grenett HE,Fuller GM. Induction and regulation of interleukin-6 gene expression in rat astrocytes. J Neuroimmunol. 1990 Dec;30(2-3):201-12.
82. Krasowska-Zoladek A,Banaszewska M,Kraszpulski M,Konat GW. Kinetics of inflammatory response of astrocytes induced by TLR 3 and TLR4 ligation. J Neurosci Res. 2007 Jan;85(1):205-12.
83. Lee SC,Dickson DW,Liu W,Brosnan CF. Induction of nitric oxide synthaseactivity in human astrocytes by interleukin-1 beta and interferon-gamma. J Neuroimmunol. 1993 Jul;46(1-2):19-24.
84. McNaught KS,Jenner P. Extracellular accumulation of nitric oxide, hydrogen peroxide,and glutamate in astrocytic cultures following glutathione depletion,complex I inhibition,and/or lipopolysaccharide-induced activation. Biochem Pharmacol. 2000 Oct 1;60(7):979-88.
85. Li Y , Ji A , Weihe E , Sch?fer MK. Cell-specific expression and lipopolysaccharide-induced regulation of tumor necrosis factor alpha (TNFalpha) and TNF receptors in rat dorsal root ganglion. J Neurosci. 2004 Oct 27;24(43):9623-31.
86. Ohtori S,Takahashi K,Moriya H,Myers RR. TNF-alpha and TNF-alpha receptor type 1 upregulation in glia and neurons after peripheral nerve injury:studies in murine DRG and spinal cord. Spine (Phila Pa 1976). 2004 May 15;29(10):1082-8.
87. Sch?fers M,Lee DH,Brors D,Yaksh TL,Sorkin LS. Increased sensitivity of injured and adjacent uninjured rat primary sensory neurons to exogenous tumor necrosis factor-alpha after spinal nerve ligation. J Neurosci. 2003 Apr 1;23(7):3028-38.
88. Xu JT,Xin WJ,Zang Y,Wu CY,Liu XG. The role of tumor necrosis factor-alpha in the neuropathic pain induced by Lumbar 5 ventral root transection in rat. Pain. 2006 Aug;123(3):306-21.
89. Ching DW,McClintock A,Beswick F. Successful treatment with low-dose thalidomide in a patient with both Beh?et's disease and complex regional pain syndrome type I:case report. J Clin Rheumatol. 2003 Apr;9(2):96-8
90. Baron R,Wasner G. Complex regional pain syndromes. Curr Pain Headache Rep. 2001 Apr;5(2):114-23.
91. Tracey DJ,Walker JS. Pain due to nerve damage:are inflammatory mediators involved? Inflamm Res. 1995 Oct;44(10):407-11.
92. Ferreira SH,Lorenzetti BB,Bristow AF,Poole S. Interleukin-1 beta as a potent hyperalgesic agent antagonized by a tripeptide analogue. Nature. 1988 Aug 25;334(6184):698-700.
93. Maier SF,Wiertelak EP,Martin D,Watkins LR. Interleukin-1 mediates the behavioral hyperalgesia produced by lithium chloride and endotoxin. Brain Res. 1993 Oct 1;623(2):321-4.
94. Copray JC,Mantingh I,Brouwer N,Biber K,Küst BM,Liem RS,Huitinga I,Tilders FJ,Van Dam AM,Boddeke HW. Expression of interleukin-1 beta in rat dorsal root ganglia. J Neuroimmunol. 2001 Aug 30;118(2):203-11.
95. Wang XF,Huang LD,Yu PP,Hu JG,Yin L,Wang L,Xu XM,Lu PH. Upregulation of type I interleukin-1 receptor after traumatic spinal cord injury in adult rats. Acta Neuropathol. 2006 Mar;111(3):220-8.
96. Wolf G,Gabay E,Tal M,Yirmiya R,Shavit Y. Genetic impairment of interleukin-1 signaling attenuates neuropathic pain,autotomy,and spontaneous ectopic neuronal activity, following nerve injury in mice. Pain. 2006 Feb;
97. Zelenka M,Sch?fers M,Sommer C. Intraneural injection of interleukin-1beta and tumor necrosis factor-alpha into rat sciatic nerve at physiological doses induces signs of neuropathic pain. Pain. 2005 Aug;116(3):257-63.
98. Inoue Y,Fujisawa M,Kawamoto S,Shoji M,Hashizume S,Fujii M,Katakura Y,Shirahata S. Effectiveness of vitamin A acetate for enhancing the production of lung cancer specific monoclonal antibodies. Cytotechnology. 1999 Sep;31(1-2):77-83.
99. Araque A,Carmignoto G,Haydon PG. Dynamic signaling between astrocytesand neurons. Annu Rev Physiol. 2001;63:795-813.
100. Anderson ES,Lennox A,Petersen SA. New opportunities for nurses in medical education:facilitating valuable community learning experiences. Nurse Educ Pract. 2004 Jun;4(2):135-42.
101. Inoue M,Hara M,Zeng XT,Hirose T,Ohnishi S,Yasukura T,Uriu T,Omori K,Minato A,Inagaki C. Neurosci Lett. An ATP-driven Cl- pump regulates Cl- concentrations in rat hippocampal neurons.1991 Dec 16;134(1):75-8.
102. Stevens B,Fields RD. Response of Schwann cells to action potentials in development. Science. 2000 Mar 24;287(5461):2267-71.
103. Dani JW,Chernjavsky A,Smith SJ. Neuronal activity triggers calcium waves in hippocampal astrocyte networks. Neuron. 1992 Mar;8(3):429-40.
104. Parpura V,Basarsky TA,Liu F,Jeftinija K,Jeftinija S,Haydon PG. Glutamate-mediated astrocyte-neuron signalling. Nature. 1994 Jun 30;369(6483):744-7.
105. Araque A,Parpura V,Sanzgiri RP,Haydon PG. Tripartite synapses: glia, the unacknowledged partner. Trends Neurosci. 1999 May;22(5):208-15.
106. Beattie EC,Stellwagen D,Morishita W,Bresnahan JC,Ha BK,Von Zastrow M,Beattie MS,Malenka RC. Control of synaptic strength by glial TNFalpha. Science. 2002 Mar 22;295(5563):2282-5.
107. Scemes E,Giaume C. Astrocyte calcium waves: what they are and what they do. Glia. 2006 Nov 15;54(7):716-25.
108. Innocenti B,Parpura V,Haydon PG. Imaging extracellular waves of glutamate during calcium signaling in cultured astrocytes. J Neurosci. 2000 Mar 1;20(5):1800-8.
109. Fellin T,Pascual O,Gobbo S,Pozzan T,Haydon PG,Carmignoto G.Neuronal synchrony mediated by astrocytic glutamate through activation of extrasynaptic NMDA receptors. Neuron. 2004 Sep 2;43(5):729-43.
110. Zonta M,Sebelin A,Gobbo S,Fellin T,Pozzan T,Carmignoto G. Glutamate-mediated cytosolic calcium oscillations regulate a pulsatile prostaglandin release from cultured rat astrocytes. J Physiol. 2003 Dec 1;553(Pt 2):407-14.
111. Gu JG,MacDermott AB. Activation of ATP P2X receptors elicits glutamate release from sensory neuron synapses. Nature. 1997 Oct 16;389(6652):749-53.
112. Jahr CE,Jessell TM. ATP excites a subpopulation of rat dorsal horn neurones. Nature. 1983 Aug 25-31;304(5928):730-3.
113. Chen CC,Akopian AN,Sivilotti L,Colquhoun D,Burnstock G,Wood JN. A P2X purinoceptor expressed by a subset of sensory neurons. Nature. 1995 Oct 5;377(6548):428-31.
114. Kennedy. P2X receptors:targets for novel analgesics? Neuroscientist. 2005 Aug;11(4):345-56.
115. Fyffe RE,Perl ER. Is ATP a central synaptic mediator for certain primary afferent fibers from mammalian skin? Proc Natl Acad Sci U S A. 1984 Nov;81(21):6890-3.
116. Werry EL,Liu GJ,Bennett MR. Glutamate-stimulated ATP release from spinal cord astrocytes is potentiated by substance P. J Neurochem. 2006 Nov;99(3):924-36.
117. Meller ST,Dykstra C,Grzybycki D,Murphy S,Gebhart GF. The possible role of glia in nociceptive processing and hyperalgesia in the spinal cord of the rat. Neuropharmacology. 1994 Nov;33(11):1471-8.
118. Magistretti PJ,Pellerin L. Astrocytes Couple Synaptic Activity to GlucoseUtilization in the Brain. News Physiol Sci. 1999 Oct;14:177-182.
119. Ji RR,Kohno T,Moore KA,Woolf CJ. Central sensitization and LTP:do pain and memory share similar mechanisms? Trends Neurosci. 2003 Dec;26(12):696-705.
120. Ikeda H,Tsuda M,Inoue K,Murase K. Long-term potentiation of neuronal excitation by neuron-glia interactions in the rat spinal dorsal horn. Eur J Neurosci. 2007 Mar;25(5):1297-306.
121. Hald A. Spinal astrogliosis in pain models:cause and effects. Cell Mol Neurobiol. 2009 Jul;29(5):609-19.
122.王国兴,王伯瑶. Toll样受体及其信号转导.细胞与分子免疫学杂志. 2002,18 (6):666-669
123. Medzhitov R,Preston-Hurlburt P,Janeway CA Jr. A human homologue of the Drosophila Toll protein signals activation of adaptive immunity.Nature. 1997 Jul 24;388(6640):394-7.
124. Chaudhary PM,Ferguson C,Nguyen V,Nguyen O,Massa HF,Eby M,Jasmin A,Trask BJ,Hood L,Nelson PS. Cloning and characterization of two Toll/Interleukin-1 receptor-like genes TIL3 and TIL4 : evidence for a multi-gene receptor family in humans. Blood. 1998 Jun 1;91(11):4020-7.
125. Takeuchi O,Kawai T,Sanjo H,Copeland NG,Gilbert DJ,Jenkins NA, Takeda K,Akira S. TLR6:A novel member of an expanding toll-like receptor family. Gene. 1999 Apr 29;231(1-2):59-65.
126. Du X,Poltorak A,Wei Y,Beutler B. Three novel mammalian toll-like receptors: gene structure, expression, and evolution. Eur Cytokine Netw. 2000 Sep;11(3):362-71.
127. Chuang T,Ulevitch RJ. Identification of hTLR10:a novel human Toll-like receptor preferentially expressed in immune cells. Biochim Biophys Acta.2001 Mar 19;1518(1-2):157-61.
128. Toll样受体及其信号转导.王国兴,王伯瑶.细胞与分子免疫学杂志. 2002,18 (6):666-669
129. Schwandner R , Dziarski R , Wesche H , Rothe M , Kirschning CJ. Peptidoglycan- and lipoteichoic acid-induced cell activation is mediated by toll-like receptor 2. J Biol Chem. 1999 Jun 18;274(25):17406-9.
130. Ozinsky A,Underhill DM,Fontenot JD,Hajjar AM,Smith KD,Wilson CB,Schroeder L,Aderem A. The repertoire for pattern recognition of pathogens by the innate immune system is defined by cooperation between toll-like receptors. Proc Natl Acad Sci U S A. 2000 Dec 5;97(25):13766-71.
131. Alexopoulou L,Holt AC,Medzhitov R,Flavell RA. Recognition of double-stranded RNA and activation of NF-kappaB by Toll-like receptor 3. Nature. 2001 Oct 18;413(6857):732-8.
132.臧丽,王建平. TLR4与糖尿病肾病的研究进展.国际病理科学与临床杂志. 2010,30(6):523-526
133. Guo LH,Schluesener HJ. The innate immunity of the central nervous system in chronic pain: the role of Toll-like receptors. Cell Mol Life Sci. 2007 May;64(9):1128-36.
134. DeLeo JA , Tanga FY , Tawfik VL. Neuroimmune activation and neuroinflammation in chronic pain and opioid tolerance/hyperalgesia. Neuroscientist. 2004 Feb;10(1):40-52.
135. Olson JK,Miller SD. Microglia initiate central nervous system innate and adaptive immune responses through multiple TLRs. J Immunol. 2004 Sep 15;173(6):3916-24.
136. Lee SJ,Lee S. Toll-like receptors and inflammation in the CNS. Curr Drug Targets Inflamm Allergy. 2002 Jun;1(2):181-91.
137. Kielian T. Toll-like receptors in central nervous system glial inflammation and homeostasis. J Neurosci Res. 2006 Apr;83(5):711-30.
138. Ebert S,Gerber J,Bader S,Mühlhauser F,Brechtel K,Mitchell TJ, Nau R. Dose-dependent activation of microglial cells by Toll-like receptor agonists alone and in combination. J Neuroimmunol. 2005 Feb;159(1-2):87-96.
139. Jack CS,Arbour N,Manusow J,Montgrain V,Blain M,McCrea E,Shapiro A,Antel JP. TLR signaling tailors innate immune responses in human microglia and astrocytes. J Immunol. 2005 Oct 1;175(7):4320-30.
140. McMahon SB,Cafferty WB,Marchand F. Immune and glial cell factors as pain mediators and modulators. Exp Neurol. 2005 Apr;192(2):444-62.
141. De Leo JA,Tawfik VL,LaCroix-Fralish ML,De Leo JA,Tawfik VL, LaCroix-Fralish ML. Pain. 2006 May;122(1-2):17-21.
142. Durrenberger PF,Facer P,Gray RA,Chessell IP,Naylor A,Bountra C,Banati RB,Birch R,Anand P. Cyclooxygenase-2 (Cox-2) in injured human nerve and a rat model of nerve injury. J Peripher Nerv Syst. 2004 Mar;9(1):15-25.
143. Ma W,Eisenach JC. Cyclooxygenase 2 in infiltrating inflammatory cells in injured nerve is universally up-regulated following various types of peripheral nerve injury. Neuroscience. 2003;121(3):691-704.
144. Durrenberger PF,Facer P,Casula MA,Yiangou Y,Gray RA,Chessell IP,Day NC,Collins SD,Bingham S,Wilson AW,Elliot D,Birch R, Anand P. Prostanoid receptor EP1 and Cox-2 in injured human nerves and a rat model of nerve injury: a time course study. BMC Neurol. 2006 Jan 4;6:1.
145. Lee JY,Sohn KH,Rhee SH,Hwang D. Saturated fatty acids,but notunsaturated fatty acids,induce the expression of cyclooxygenase-2 mediated through Toll-like receptor 4. J Biol Chem. 2001 May 18;276(20):16683-9.
146. Yeo SJ,Yoon JG,Yi AK. Myeloid differentiation factor 88-dependent post-transcriptional regulation of cyclooxygenase-2 expression by CpG DNA:tumor necrosis factor-alpha receptor-associated factor 6,a diverging point in the Toll-like receptor 9-signaling. J Biol Chem. 2003 Oct 17;278(42):40590-600.
147. Villamón E,Roig P,Gil ML,Gozalbo D. Toll-like receptor 2 mediates prostaglandin E(2) production in murine peritoneal macrophages and splenocytes in response to Candida albicans. Res Microbiol. 2005 Jan-Feb;156(1):115-8.
148. Hosoi T,Suzuki S,Nomura J,Ono A,Okuma Y,Akira S,Nomura Y. Bacterial DNA induced iNOS expression through MyD88-p38 MAP kinase in mouse primary cultured glial cells. Brain Res Mol Brain Res. 2004 May 19;124(2):159-64
149. Kakimura J,Kitamura Y,Takata K,Umeki M,Suzuki S,Shibagaki K,Taniguchi T,Nomura Y,Gebicke-Haerter PJ,Smith MA,Perry G, Shimohama S. Microglial activation and amyloid-beta clearance induced by exogenous heat-shock proteins. FASEB J. 2002 Apr;16(6):601-3.
150. Bussini S,Meda L,Scarpini E,Clementi E,Conti G,Tiriticco M,Bresolin N,Baron P. Heparan sulfate proteoglycan induces the production of NO and TNF-alpha by murine microglia. Immun Ageing. 2005 Jul 16;2:11.
151. Hwang D. Modulation of the expression of cyclooxygenase-2 by fatty acids mediated through toll-like receptor 4-derived signaling pathways. FASEB J. 2001 Dec;15(14):2556-64.
152. Takeda K,Akira S. Toll-like receptors in innate immunity. Int Immunol.2005 Jan;17(1):1-14.
153. Noubir S,Hmama Z,Reiner NE. Dual receptors and distinct pathways mediate interleukin-1 receptor-associated kinase degradation in response to lipopolysaccharide. Involvement of CD14/TLR4 , CR3 , and phosphatidylinositol 3-kinase. J Biol Chem. 2004 Jun 11;279(24):25189-95.
154. Dell'Antonio G,Quattrini A,Dal Cin E,Fulgenzi A,Ferrero ME. Antinociceptive effect of a new P(2Z)/P2X7 antagonist,oxidized ATP,in arthritic rats. Neurosci Lett. 2002 Jul 19;327(2):87-90.
155. Tsuda M,Shigemoto-Mogami Y,Koizumi S,Mizokoshi A,Kohsaka S,Salter MW,Inoue K. P2X4 receptors induced in spinal microglia gate tactile allodynia after nerve injury. Nature. 2003 Aug 14;424(6950):778-83.
156. Trang T,Beggs S,Salter MW. Purinoceptors in microglia and neuropathic pain. Pflugers Arch. 2006 Aug;452(5):645-52.
157. Nasu-Tada K,Koizumi S,Tsuda M,Kunifusa E,Inoue K. Possible involvement of increase in spinal fibronectin following peripheral nerve injury in upregulation of microglial P2X4,a key molecule for mechanical allodynia. Glia. 2006 May;53(7):769-75.
158. Guo LH,Guo KT,Wendel HP,Schluesener HJ. Combinations of TLR and NOD2 ligands stimulate rat microglial P2X4R expression. Biochem Biophys Res Commun. 2006 Oct 27;349(3):1156-62.
159. Kahlenberg JM,Lundberg KC,Kertesy SB,Qu Y,Dubyak GR. Potentiation of caspase-1 activation by the P2X7 receptor is dependent on TLR signals and requires NF-kappaB-driven protein synthesis. J Immunol. 2005 Dec 1;175(11):7611-22.
160. Bachmann M,Horn K,Poleganov MA,Paulukat J,Nold M, Pfeilschifter J,Mühl H. Interleukin-18 secretion and Th1-like cytokine responses inhuman peripheral blood mononuclear cells under the influence of the toll-like receptor-5 ligand flagellin. Cell Microbiol. 2006 Feb;8(2):289-300.
161. Tanga FY,Nutile-McMenemy N,DeLeo JA. The CNS role of Toll-like receptor 4 in innate neuroimmunity and painful neuropathy. Proc Natl Acad Sci U S A. 2005 Apr 19;102(16):5856-61.
162. Raghavendra V,Tanga F,DeLeo JA. Inhibition of microglial activation attenuates the development but not existing hypersensitivity in a rat model of neuropathy. J Pharmacol Exp Ther. 2003 Aug;306(2):624-30.
163. Watkins LR,Maier SF. Immune regulation of central nervous system functions: from sickness responses to pathological pain. J Intern Med. 2005 Feb;257(2):139-55.
164. Inoue K. The function of microglia through purinergic receptors : neuropathic pain and cytokine release. Pharmacol Ther. 2006 Jan;109(1-2):210-26
165. Ledeboer A,Sloane EM,Milligan ED,Frank MG,Mahony JH,Maier SF,Watkins LR. Minocycline attenuates mechanical allodynia and proinflammatory cytokine expression in rat models of pain facilitation. Pain. 2005 May;115(1-2):71-83.
166. Marchand F,Perretti M,McMahon SB. Role of the immune system in chronic pain. Nat Rev Neurosci. 2005 Jul;6(7):521-32
167. Lehnardt S,Massillon L,Follett P,Jensen FE,Ratan R,Rosenberg PA, Volpe JJ,Vartanian T. Activation of innate immunity in the CNS triggers neurodegeneration through a Toll-like receptor 4-dependent pathway. Proc Natl Acad Sci U S A. 2003 Jul 8;100(14):8514-9.
168. Tanga FY,Raghavendra V,DeLeo JA. Quantitative real-time RT-PCR assessment of spinal microglial and astrocytic activation markers in a ratmodel of neuropathic pain. Neurochem Int. 2004 Jul-Aug;45(2-3):397-407.
169. Zhang Z,Trautmann K,Schluesener HJ. Spinal cord glia activation following peripheral polyinosine-polycytidylic acid administration. Neuroreport. 2005 Sep 8;16(13):1495-9.
170. Guo LH,Schluesener HJ. Acute but not chronic stimulation of glial cells in rat spinal cord by systemic injection of lipopolysaccharide is associated with hyperalgesia. Acta Neuropathol. 2006 Dec;112(6):703-13.
171. Zhang Z,Trautmann K,Schluesener HJ. Microglia activation in rat spinal cord by systemic injection of TLR3 and TLR7/8 agonists. J Neuroimmunol. 2005 Jul;164(1-2):154-60.
172. Su Y,Zhang Z,Trautmann K,Xu S,Schluesener HJ. TLR and NOD2 ligands induce cell proliferation in the rat intact spinal cord. J Neuropathol Exp Neurol. 2005 Nov;64(11):991-7.
173. Rivest S. Molecular insights on the cerebral innate immune system. Brain Behav Immun. 2003 Feb;17(1):13-9.
174. Watkins LR,Hutchinson MR,Johnston IN,Maier SF. Glia:novel counter-regulators of opioid analgesia. Trends Neurosci. 2005 Dec;28(12):661-9.
175. Johnston IN, Westbrook RF. Acute and conditioned sickness reduces morphine analgesia. Behav Brain Res. 2003 Jun 16;142(1-2):89-97.
176. Johnston IN,Westbrook RF. Inhibition of morphine analgesia by LPS: role of opioid and NMDA receptors and spinal glia. Behav Brain Res. 2005 Jan 6;156(1):75-83.
177. Sun Y,Cai TT,Shen Y,Zhou XB,Chen T,Xu Q. Si-Ni-San,a traditional Chinese prescription , and its active ingredient glycyrrhizin ameliorate experimental colitis through regulating cytokine balance. IntImmunopharmacol. 2009 Nov; 9(12):1437-43.
178. Adler G,Schmid RM. Chronic pancreatitis: still puzzling? Gastroenterology. 1997; 112(5):1762-5.
179. Feng QX,Wang W,Feng XY,Mei XP,Zhu C,Liu ZC,Li YQ,Dou KF,Zhao QC:Astrocytic activation in thoracic spinal cord contributes to persistent pain in rat model of chronic pancreatitis. Neuroscience 2010, 167:501-509.
180. Wang W,Wang W,Wang Y,Huang J,Wu S,Li YQ. Temporal changes of astrocyte activation and glutamate transporter-1 expression in the spinal cord after spinal nerve ligation-induced neuropathic pain. Anat Rec (Hoboken). 2008 May;291(5):513-8.
181. Mei X,Wang W,Wang W,Li Y,Zhang H,Wu S,Li Y,Xu L. Inhibiting astrocytic activation:a novel analgesic mechanism of ketamine at the spinal level? J Neurochem. 2009 Jun;109(6):1691-700.
182. Raghavendra V,Tanga FY,DeLeo JA. Attenuation of morphine tolerance, withdrawal-induced hyperalgesia , and associated spinal inflammatory immune responses by propentofylline in rats. Neuropsychopharmacology. 2004 Feb;29(2):327-34.
183. Zhang RX,Liu B,Wang L,Ren K,Qiao JT,Berman BM,Lao L. Spinal glial activation in a new rat model of bone cancer pain produced by prostate cancer cell inoculation of the tibia. Pain. 2005 Nov;118(1-2):125-36.
184. Saab CY,Wang J,Gu C,Garner KN,Al-Chaer ED. Microglia: a newly discovered role in visceral hypersensitivity? Neuron Glia Biol. 2007 Apr 13;2(4):271-277.
185. Watkins LR,Milligan ED,Maier SF. Glial activation:a driving force for pathological pain. Trends Neurosci. 2001 Aug;24(8):450-5.
186. Zhuang ZY,Gerner P,Woolf CJ,Ji RR. ERK is sequentially activated in neurons, microglia, and astrocytes by spinal nerve ligation and contributes to mechanical allodynia in this neuropathic pain model. Pain. 2005 Mar;114(1-2):149-59.
187. Watkins LR,Hutchinson MR,Ledeboer A,Wieseler-Frank J,Milligan ED,Maier SF. Norman Cousins Lecture. Glia as the "bad guys": implications for improving clinical pain control and the clinical utility of opioids. Brain Behav Immun. 2007 Feb;21(2):131-46.
188. Ceyhan GO,Bergmann F,Kadihasanoglu M,Altintas B,Demir IE,Hinz U,Müller MW,Giese T,Büchler MW,Giese NA,Friess H. Pancreatic neuropathy and neuropathic pain--a comprehensive pathomorphological study of 546 cases. Gastroenterology. 2009 Jan;136(1):177-186.e1.
189. Aicher SA,Sharma S,Cheng PY,Pickel VM. The N-methyl-D-aspartate (NMDA) receptor is postsynaptic to substance P-containing axon terminals in the rat superficial dorsal horn. Brain Res. 1997 Oct 24;772(1-2):71-81.
190. Araque A,Parpura V,Sanzgiri RP,Haydon PG. Glutamate-dependent astrocyte modulation of synaptic transmission between cultured hippocampal neurons. Eur J Neurosci. 1998 Jun;10(6):2129-42.
191. Watkins LR,Milligan ED,Maier SF. Spinal cord glia:new players in pain. Pain. 2001 Sep;93(3):201-5. Watkins LR, Milligan ED, Maier SF. Glial activation:a driving force for pathological pain. Trends Neurosci. 2001 Aug;24(8):450-5.
192. Wieseler-Frank J,Maier SF,Watkins LR. Central proinflammatory cytokines and pain enhancement. Neurosignals. 2005;14(4):166-74.
193. Guo W,Wang H,Watanabe M,Shimizu K,Zou S,LaGraize SC,Wei F,Dubner R,Ren K. Glial-cytokine-neuronal interactions underlying themechanisms of persistent pain. J Neurosci. 2007 May 30;27(22):6006-18.
194. Gao YJ,Zhang L,Samad OA,Suter MR,Yasuhiko K,Xu ZZ,Park JY,Lind AL,Ma Q,Ji RR. JNK-induced MCP-1 production in spinal cord astrocytes contributes to central sensitization and neuropathic pain. J Neurosci. 2009 Apr 1;29(13):4096-108
195. Wang W,Wang W,Mei X,Huang J,Wei Y,Wang Y,Wu S,Li Y. Crosstalk between spinal astrocytes and neurons in nerve injury-induced neuropathic pain. PLoS One. 2009 Sep 11;4(9):e6973.
196. Tiscornia OM,Martinez JL,Sarles H. Some aspects of human and canine macroscopic pancreas innervation. Am J Gastroenterol. 1976 Oct;66(4):353-61.
197. Schloithe AC,Sutherland K,Woods CM,Blackshaw LA,Davison JS,Toouli J,Saccone GT. A novel preparation to study rat pancreatic spinal and vagal mechanosensitive afferents in vitro. Neurogastroenterol Motil. 2008 Sep;20(9):1060-9.
198. Hutchinson MR,Lewis SS,Coats BD,Rezvani N,Zhang Y,Wieseler JL,Somogyi AA,Yin H,Maier SF,Rice KC,Watkins LR:Possible involvement of toll-like receptor 4/myeloid differentiation factor-2 activity of opioid inactive isomers causes spinal proinflammation and related behavioral consequences. Neuroscience 2010,167:880-893.
199. Bettoni I,Comelli F,Rossini C,Granucci F,Giagnoni G,Peri F, Costa B:Glial TLR4 receptor as new target to treat neuropathic pain:efficacy of a new receptor antagonist in a model of peripheral nerve injury in mice. Glia 2008,56:1312-1319.
200. Obata K,Katsura H,Miyoshi K,Kondo T,Yamanaka H,Kobayashi K,Dai Y,Fukuoka T,Akira S,Noguchi K:Toll-like receptor 3 contributes tospinal glial activation and tactile allodynia after nerve injury. J Neurochem. 2008 Jun 1;105(6):2249-59.
201. Kim D,Kim MA,Cho IH,Kim MS,Lee S,Jo EK,Choi SY,Park K,Kim JS,Akira S,Na HS,Oh SB,Lee SJ. A critical role of toll-like receptor 2 in nerve injury-induced spinal cord glial cell activation and pain hypersensitivity. J Biol Chem 2007,282:14975-14983.
202. Cao L,Tanga FY,Deleo JA: The contributing role of CD14 in toll-like receptor 4 dependent neuropathic pain. Neuroscience 2009,158:896-903.
203. Bsibsi M,Ravid R,Gveric D,van Noort JM:Broad expression of Toll-like receptors in the human central nervous system. J Neuropathol Exp Neurol 2002,61:1013-1021.
204. McKimmie CS,Fazakerley JK:In response to pathogens, glial cells dynamically and differentially regulate Toll-like receptor gene expression. J Neuroimmunol 2005,169:116-125
205. McKimmie CS,Johnson N,Fooks AR,Fazakerley JK:Viruses selectively upregulate Toll-like receptors in the central nervous system. Biochem Biophys Res Commun 2005,336:925-933.
206. Carpentier PA,Begolka WS,Olson JK,Elhofy A, Karpus WJ,Miller SD:Differential activation of astrocytes by innate and adaptive immune stimuli. Glia 2005, 49:360-374.
207. Wang W,Wang YY,Huang J,Wu SX,Li YQ:Temporal changes of astrocyte activation and glutamate transporter-1 expression in the spinal cord after spinal nerve ligation-induced neuropathic pain. Anat Rec (Hoboken) 2008,291:513-518.
208. Akira S,Takeda K,Kaisho T:Toll-like receptors:critical proteins linking innate and acquired immunity. Nat Immunol 2001,2:675-680.
209. Bsibsi M,Persoon-Deen C,Verwer RW,Meeuwsen S,Ravid R,Van Noort JM:Toll-like receptor 3 on adult human astrocytes triggers production of neuroprotective mediators. Glia 2006, 53:688-695.
210. Park C,Lee S,Cho IH,Lee HK,Kim D,Choi SY,Oh SB,Park K, Kim JS,Lee SJ:TLR3-mediated signal induces proinflammatory cytokine and chemokine gene expression in astrocytes: differential signaling mechanisms of TLR3-induced IP-10 and IL-8 gene expression. Glia 2006, 53:248-256.
211. Zhang RX,Li A,Liu B,Wang L,Ren K,Zhang H,Berman BM,Lao L:IL-1ra alleviates inflammatory hyperalgesia through preventing phosphorylation of NMDA receptor NR-1 subunit in rats. Pain 2008, 135:232-239.
212. Choi JI,Svensson CI,Koehrn FJ,Bhuskute A,Sorkin LS:Peripheral inflammation induces tumor necrosis factor dependent AMPA receptor trafficking and Akt phosphorylation in spinal cord in addition to pain behavior. Pain. 2010 May;149(2):243-53.
213. Dominguez E , Mauborgne A , Mallet J , Desclaux M , Pohl M : SOCS3-mediated blockade of JAK/STAT3 signaling pathway reveals its major contribution to spinal cord neuroinflammation and mechanical allodynia after peripheral nerve injury. J Neurosci 2010,30:5754-5766
214. Vardeh D,Wang D,Costigan M,Lazarus M,Saper CB,Woolf CJ, Fitzgerald GA,Samad TA:COX2 in CNS neural cells mediates mechanical inflammatory pain hypersensitivity in mice. J Clin Invest 2009,119:287-294.
215. Puig-Divi V,Molero X,Salas A,Guarner F,Guarner L,Malagelada JR: Induction of chronic pancreatic disease by trinitrobenzene sulfonic acid infusion into rat pancreatic ducts. Pancreas 1996,13:417-424.