肾阳虚证动物模型造模方法综述

肾阳虚证动物模型是建立早的中医证候动物模型,20世纪60年代初,邝安等开创了中医动物模型的研究,第一次将科学实验方法引入传统中医药学的研究领域,成为中医药现代化发展方向的新的突破口.下面就笔者收集的关于肾阳虚证动物模型的造模方法按照造模依据加以简单的回顾.

　　A great deal of research work about acupuncture in terms of modern medicine has been done in China. The initial investigation about the mechanism of the clinical effect of acupuncture in the fifties, the extensive clinical practice with acupuncture in the mid-sixties, and the large scale of experimental studies on animal models about acupuncture in the seventies, especially the nineties, which have all provided many new progresses. And the following is an overview of the achievements of these research work.

药物筛选模型研究进展

药物筛选模型（drug screening model, drug screening assay method）是用于证明某种物质具有药理活性(生物活性、治疗作用)的实验方法，这些实验方法是寻找和发现药物的重要条件之一。人们在长期寻找药物的实践过程中，建立了大量用于新药筛选的各类模型，在新药发现和研究中发挥了积极作用。随着生命科学的发展，新的药物筛选模型不断出现，这些新的筛选模型不仅促进了药物的发现，而且对药物筛选的方法、理论、技术都产生了巨大影响［1］。本文对近年来药物筛选模型的研究进展作简单综述。

动物模型在腹膜透析相关研究中的应用进展

随着腹膜透析技术的普及和发展,腹膜溶质交换、局部液体转运、腹腔内药代动力学、腹膜透析液生物相容性、腹膜透析并发症等腹膜透析相关领域的研究受到肾脏病学者和医师的高度关注,而腹膜透析动物模型的应用为上述领域的体内研究提供了现实可行的手段,深化了人们对于这一肾脏替代治疗方式的认识.

食物过敏动物模型中T调节细胞的功能及活化基因foxp3、肿瘤坏死因子受体mRNA表达研究

目前有大量研究证实CD4+CD25+T调节性细胞参与了过敏性疾病,其免疫调节作用已在许多免疫性疾病动物模型中得到证实[1].

We previously found that the K141N mutation in heat shock protein B8 (HSPB8) was respon-sible for Charcot-Marie-Tooth disease type 2L in a large Chinese family. The objective of the present study was to generate a transgenic mouse model bearing the K141N mutation in the human HSPB8 gene, and to determine whether this K141NHSPB8 transgenic mouse model would manifest the clinical phenotype of Charcot-Marie-Tooth disease type 2L, and consequently be suitable for use in studies of disease pathogenesis. Transgenic mice overexpressing K141NHSPB8 were generated using K141N mutant HSPB8 cDNA cloned into a pCAGGS plasmid driven by a human cytomegalovirus expression system. PCR and western blot analysis conifrmed integra-tion of the K141NHSPB8 gene and widespread expression in tissues of the transgenic mice. The K141NHSPB8 transgenic mice exhibited decreased muscle strength in the hind limbs and impaired motor coordination, but no obvious sensory disturbance at 6 months of age by behavioral assess-ment. Electrophysiological analysis showed that the compound motor action potential amplitude in the sciatic nerve was signiifcantly decreased, but motor nerve conduction velocity remained normal at 6 months of age. Pathological analysis of the sciatic nerve showed reduced myelinated ifber density, notable axonal edema and vacuolar degeneration in K141NHSPB8 transgenic mice, suggesting axonal involvement in the peripheral nerve damage in these animals. These ifndings indicate that the K141NHSPB8 transgenic mouse successfully models Charcot-Marie-Tooth disease type 2L and can be used to study the pathogenesis of the disease.

Fluid percussion-induced traumatic brain injury models have been widely used in experimental research for years. In an experiment, the stability of impaction is inevitably affected by factors such as the appearance of liquid spikes. Management of impact pressure is a crucial factor that determines the stability of these models, and direction of impact control is another basic ele-ment. To improve experimental stability, we calculated a pressure curve by generating repeated impacts using a lfuid percussion device at different pendulum angles. A stereotactic frame was used to control the direction of impact. We produced stable and reproducible models, including mild, moderate, and severe traumatic brain injury, using the MODEL01-B device at pendulum angles of 6°, 11° and 13°, with corresponding impact force values of 1.0 ± 0.11 atm (101.32 ± 11.16 kPa), 2.6 ± 0.16 atm (263.44 ± 16.21 kPa), and 3.6 ± 0.16 atm (364.77 ± 16.21 kPa), respec-tively. Behavioral tests, hematoxylin-eosin staining, and magnetic resonance imaging revealed that models for different degrees of injury were consistent with the clinical properties of mild, moderate, and severe craniocerebral injuries. Using this method, we established lfuid percussion models for different degrees of injury and stabilized pathological features based on precise power and direction control.

Experimental al ergic encephalomyelitis is a mouse model of human multiple sclerosis with similar pathology and pathogenesis. Th1 cells play an important role in the pathogenesis of experimental al ergic encephalomyelitis. This study determined the potential effect of programmed celldeath 1 ligand 1 in the pathogenesis of experimental al ergic encephalomyelitis induced by injecting myelin oligodendrocyte glycoprotein, complete Freund’s adjuvant and Bordetel a pertussis toxin into C57BL/6J mice. Experimental al ergic encephalomyelitis mice developed disease and showed in-flammatory changes in the central nervous system by hematoxylin-eosin staining of spinal cord pathological sections, demyelination by Luxol fast-blue staining and clinical manifestations. The expression of programmed celldeath 1 ligand 1 in mice was detected by immunohistochemistry, flow cytometry and western blot analysis. The expression of programmed celldeath 1 ligand 1 in the spinal cord and splenocytes of mice was significantly increased compared with normal mice. Our findings suggest the involvement of programmed celldeath 1 ligand 1 in the pathogenesis of expe-rimental al ergic encephalomyelitis and suggest this should be studied in multiple sclerosis.

Spastic cerebral palsy is general y considered to result from cerebral cortical or pyramidal tract damage. Here, we precisely targeted the left pyramidal tract of 2-month-old Sprague-Dawley rats placed on a stereotaxic instrument under intraperitoneal anesthesia. Based on the rat brain ste-reotaxic map, a 1-mm hole was made 10 mm posterior to bregma and 0.8 mm left of sagittal suture. A microsyringe was inserted perpendicularly to the surface of the brain to a depth of 9.7 mm, and 15μL of ethanol was slowly injected to establish a rat model of spastic cerebral palsy. After modeling, the rats appeared to have necrotic voids in the pyramidal tract and exhibited typical signs and symptoms of flexion spasms that lasted for a long period of time. These findings indicate that this is an effective and easy method of establishing a rat model of spastic cerebral palsy with good re-producibility. Ethanol as a chemical ablation agent specifical y and thoroughly damages the pyra-midal tract, and therefore, the animals display flexion spasms, which are a typical symptom of the disease.

高压氧对动物有机磷中毒后神经损害恢复的实验研究

Objective To investigate the effect of hyperbarci oxygen(HBO) on recovery of nerves injury in rats suffered from acute organophosphorus poisoning. Method We established organophosphorus poisoning models and observed effect of HBO on recovery of injure nerves. Results Compared with control group, cerebrospinal fluid induced peak potential and incubation period in HBO group were significantly recovered(P<0.05).HBO could accelerated repair of injured nerves. Conclusion HBO could relieve injury of nerves during treatment of organophosphorus poisoning. ``

一种新的兔株网膜下腔出血后症状性脑血管痉挛模型的建立

Objective To establish an experimental model of symptomatic cerebral vasospasm(CVS) after subarachnoid hemorrhage( SAH) in rabbits. Method 2 weeks after the ligation of bilateral common carotid arteries, We induced CVS by injecting arterial blood twice via a cranial hole 2 mm× 2 mm and then neurological symptoms ,cerebral blood flow(rCBF) and food intake were evaluated. Results Food intake and rCBF decreased and neurological disorders were observed. Conclusion An experimental rabbit model of symptomatic CVS can be established by injecting blood via a cranial hole after bilateral common carotid arteries ligation. ``

肝衰竭模型研究进展

肝衰竭(HF)病因主要包括病毒、药物、血流动力学异常、代谢异常等[1-2].研究合适HF的模型不仅可以更好的理解HF的病理生理学状态,还可为发展和改进HF的治疗措施提供条件.在过去30年中,已发展了多种动物模型,多为急性肝衰竭(FHF)模型,方法主要包括外科手术、药物和基因敲除.近年来,有学者用肝衰竭患者血浆培养HepG2,C3A,HHY41,猪肝,成人肝等细胞.观察其对细胞影响,方法多集中于对细胞生长和活力的影响,抑制细胞大分子生物合成,对肝细胞凋亡及肝细胞解毒作用的影响.

隐孢子虫的体外培养和感染动物模型的研究进展

隐孢子虫(Cryptosporidium)于1907年由Tyzzer[1]在实验小鼠胃内首次发现并描述,而此后的70年中并未得到医学及兽医学界的关注.1976年Nime等[2]和Meisel[3]相继报道了一名免疫功能正常的儿童和一名免疫抑制的成人的隐孢子虫病病例.1982年以来,随着艾滋病(AIDS)的出现和流行,对本虫的流行病学、诊断和治疗的研究明显增多.隐孢子虫属属于隐孢子虫科(Cryptosporidiids),艾美球虫亚目(Einerioiine),真球虫目(Eucoccidiorida),球虫亚纲(Asin Coccidia),孢子虫纲(Sporozoasida),端复亚门(Apicanplex).公认有6个种:小鼠隐孢子虫(C.muris)、微小隐孢子虫(C.parvum)、火鸡隐孢子虫(C.meleagridis)、贝氏隐孢子虫(C.baileyi)、鱼隐孢子虫(C.nasorum)、蛇隐孢子虫(C.serpentis).人的隐孢子虫病(cryptosporidiosis)是由C.parvum引起.隐孢子虫为一种重要的引起人和动物腹泻的机会性致病原虫.隐孢子虫能够感染多种脊椎动物并导致相应疾病,通常会引起哺乳动物稀水样腹泻便、鸟类的腹泻或呼吸道症状和鱼类及两栖类的胃炎[4],为人兽共患寄生虫病.

It is well known that the arterial baroreflex(ABR)plays a key role in the regulation of heart rate and stabilization of blood pressure.Currently,it appears that ABR dysfunction is involved in the pathophysiology of cardiovascular disease states.Since the mid-1990s,a number of studies have been carried out in our laboratory to explore the pathological significance of ABR function in cardiovascular damage.This minireview summarizes our research work on the topic of ABR and left ventricular hypertrophy(LVH).On the basis of discussion concerning the importance of ABR dysfunction in hypertensive LVH and sinoaortic denervation-induced LVH,we advance a new strategy for reversal of LVH,that is,restoration of impaired ABR function.We tested this hypothesis in animal models with ABR deficiency.It was found that improvement of impaird ABR function with long-term treatment of ketanserin or candesartan was accompanied by reversal of LVH.The preliminary results indicate that it is feasible to target ABR for treatment of LVH.

Sepsis and septic shock have continued to produce significant morbidity and mortality, in recent times, in acutely injured patients as well as patients in the intensive care units despite advances in antibiotic therapy and in the cardi ovascular and pulmonary support for these patients. This emphasizes the need to gain a better understanding of the fundamental mechanisms of the pathogenesis of sepsis syndrome in the critically ill or injured patients. The present knowled ge of the mechanisms of septic pathogenesis clearly indicates involvement of mod ulations in the functions of the cells of body's immune defense system, namely, monocytes/macrophages, polymorphonuclear leukocytes, and lymphocytes. Most of su ch knowledge has been derived from laboratory experiments in animal models of se ptic injury, or from studies of immune-system cells, in vitro. Clinical stud ies have also supported the role of immune perturbations. Yet, to date, very few the rapeutic approaches are available to effectively counter the sepsis-related immu ne disturbances. Functional modulations in the immune system cells, in the injured/septic hos ts, can exert not only adaptive/beneficial effects but also profoundly adverse effects on the non-immune-system cells such as endothelial, epithelial, neurona l, endocrine, neuro-endocrine, smooth muscle, skeletal muscle, and cardiac muscl e cells. The primary functional modulation in the immune-system cells after inju ry or with critical illness is activation of such cells via molecules from pat hogens, for example, lipopolysaccharide from gram negative organisms, lipoteic hoic acid/peptidoglycan from gram positive organisms, or zymosan from fungi. Suc h pathogenic molecules activate monocytes and tissue macrophages to result in th e expression and release of cytokine mediators (TNFα, IL-1, IL-6, IL-8, and IL- 10), as well as certain lipid mediators (PGE2, LTB4, PAF). While these media tors could play a host-defense role in support of the host via containment/destr uctio n of the pathogens, they could also exert detrimental effects in the host and co ntribute to host morbidity and mortality. Some of these mediators (TNFα, IL-1, IL-6, IL-8, LTB4, PAF) have been shown to be “pro-inflammatory”,and to potenti al ly exert a harmful effect on non-immune cell systems such as endothelial cells, epithelial cells, and muscle cells. Among the pro-inflammatory mediators, TNFα a nd IL-1 could play major harmful roles, and thus contribute to the injured host morbidity and mortality. Mediators, IL-10 and PGE2 have been shown to be “an ti-inflammatory” and to potentially contribute to a dreadful state of immuno-s uppression in the injured hosts, which can also lead to morbidity and mortality. Whi ch is worse, a harmful pro-inflammatory phase or harmful immuno-suppression ? Or Which occurs first, a harmful pro-inflammatory condition or a harmful immuno-su ppression? These are questions which can not be definitively answered for the g eneral population of critically ill/injured patients. It is reasonable to assum e that the answers to these question would vary from one patient subset to anot her patient subset. Thus, whether to treat the patient with a putative anti-pro- inflammatory agent or with a putative anti-anti-inflammatory agent remai ns unresolved for the general sepsis patient population. Paradoxically, TNFα, IL-1, and other pro-inflammatory mediators under certa in circumstances may serve as natural “blockers” of immuno-suppression or “pr omoters” of immune stimulation. This may be true in the case of some of the se ptic patients. Understandably, these patients should not be treated with anti-pro-inf lammatory agents. On the other hand, the anti-inflammatory mediators such as PGE 2, IL-10, and certain other naturally occurring antagonists of TNFα and IL-10 a ctions (TNFα, and IL-1 receptor antagonists) could not only be producing a cert ai n level of immune-suppression but also serving as important feed back controller of the pro-inflammatory mediators. Thus some of the naturally occurring anti-I nflammatory agents could indeed serve as adaptive/beneficial “anti-pro-inflamma tory”, therapeutic agents in certain subset of sepsis patients. Although there is little doubt that effective therapeutic control of the sep sis syndrome could be achieved via appropriate modulation of the cells of the I mmune system, at the present time we do not have an immune therapeutic regimen w hich can singly be efficacious for the general population of patients with the s eptic complication. This implies that before an effective treatment of sepsis p atients, the patients must be identified, by some diagnostic procedure, as to wh ether they need an anti-pro-inflammatory therapy or an anti-anti-inflammatory th erapy. Thus, although immuno-therapy of sepsis remains promising, its efficacy a waits further investigative work particularly through clinical studies in sepsis patients.

AIM:The direct renin inhibitor aliskiren displays antihypertensive and antialbuminuric effects in humans and in animal models . Emerging evidence has shown that aliskiren localizes and persists in medullary collecting ducts even after treatment was discontinued . The purpose of the present study was to investigate whether aliskiren regulates renal aquaporin expression and improves urinary concen -trating defect induced by lithium .METHODS:The mice were either fed with normal chow or LiCl diet (40 mmol/kg dry food per day for first 4 days and 20 mmol/kg dry food per day for last 3 days ) for seven days .Some mice were intraperitoneally injected aliskiren ( 50 mg/kg BW per day in saline ) .RESULTS:Mice injected aliskiren developed decreased urine output and increased urine osmolal -ity when compared with controls .Aliskiren significantly increased protein abundance of AQP 2 and phosphorylated-S256 AQP2 in the kidney inner medulla .Immunohistochemistry and immunofluoresence showed increased apical and intracellular labeling of AQP 2 and pS256-AQP2 in collecting duct principal cells of kidneys in mice treated with aliskiren .Aliskiren treatment prevented urinary concen-trating defect in lithium-treated mice , and improved the downregulation of AQP 2 and pS256-AQP2 protein abundance in inner medulla of the kidney .In primary cultured rat inner medulla collecting duct cells , aliskiren dramatically increased AQP 2 protein abundance which was significantly inhibited either by PKA inhibitor H 89 or by adenylyl cyclase inhibitor MDL 12330, indicating an involvement of the cAMP signalling pathway in mediating aliskiren-induced increased AQP 2 expression .CONCLUSION: The direct renin inhibitor aliskiren upregulates AQP 2 protein expression in inner medullary collecting duct principal cells and prevents lithium -induced nephro-genic diabetes insipidus ( NDI) likely via PKA-cAMP pathways .