Fatty acid and amino acid composition in haruan as a potential role in wound healing

1. Two species of snakehead fish are available in Sabah, i.e. Channa striatus and Channa melanosoma, and are commonly known as haruan. Haruan is consumed by many Malaysians to induce healing after a clinical operations. However, there is no scientific evidence as yet to substantiate the claim, and so it was decided to analyse the biochemical composition in haruan to determine which compounds may have a possible role or potential in wound healing.2. Samples (midline fillet) of both species were extracted separately in hexane for the qualitative analysis of fatty acids by a gas chromatography, Hewlett-Packard 5890A, using a 10 meter superox 11 column (Alltech) at temperature between 190 and 245°C. Peak areas were calculated automatically using Hewlett-Packard 3393A computing integrator. Subsequently, the amino acid composition was analysed using a precolumn derivatization reverse phase HPLC waters PICO-TAG system.3. Haruan is found to contain unusually high arachidonic acid (AA) but almost no eicosapentaenoic acid (EPA). AA which is a precursor of prostaglandin may initiate blood clotting and be responsible for growth. Haruan also contains all the essential amino acids for wound healing, particularly glycine which is the most important component of human skin collagen. Therefore, haruan contained all the basic biochemical requirements for wound healing.     

Effects of icariin on the fracture healing in young and old rats and its mechanism

ContextIcariin has attracted increasing attention because of its wide variety of pharmacological effects.ObjectiveThis study investigates whether icariin could promote fracture healing in young and old rats and its mechanisms.Materials and methodsA Wistar rat model for the tibia fracture in relatively young and old rats, respectively, was established. The rats were divided into four groups: model group, L-icariin (50 mg/kg icariin), M-icariin (100 mg/kg icariin) and H-icariin (200 mg/kg icariin), and intragastric administration of icariin was performed for 10 days or 20 days. In addition, isolated and cultured rat bone mesenchymal stem cells (rBMSCs) from young and old rats were cultured with 5% and 20% of icariin-containing serum, respectively, then cell viability and alkaline phosphatase (ALP) activity were measured.ResultsIcariin administration induced the expression of Runx2, Osterix, BMP-2, p-Smad5 and osteocalcin secretion (young rats: model: 2.50 ± 0.71; L-icariin: 10.10 ± 1.55; M-icariin: 24.95 ± 2.19; H-icariin: 36.80 ± 2.26; old rats: model: 1.55 ± 0.49; L-icariin:6.55 ± 0.50; M-icariin: 15.00 ± 0.85; H-icariin:20.50 ± 2.27) at the fracture site, and increased the levels of bone formation markers (OC, BAP, NTX-1 and CTX-1) in a dose-dependent manner. In vitro, icariin treatment promoted rBMSC viability, increased ALP activity and the expression of BMP-2/Smad5/Runx2 pathway proteins.Discussion and conclusionsIcariin may accelerate fracture healing by activating the BMP-2/Smad5/Runx2 pathway in relatively young and old rats. The research on the mechanism of icariin to promote fracture healing can provide a theoretical basis for the clinical application and promotion of icariin.     

BCH, an inhibitor of system L amino acid transporters, induces apoptosis in cancer cells

PURPOSE: L-Type amino acid transporter 1 (LAT1) is highly expressed in cancer cells to support their continuous growth and proliferation. We have examined the effect of 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH), an inhibitor of system L amino acid transporters, and the mechanism by which BCH suppresses cell growth in cancer cells. METHODS: The effect of BCH and the mechanism of BCH on cell growth suppression in cancer cells were examined using amino acid transport measurement, MTT assay, DNA fragmentation analysis, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay and immunoblotting. RESULTS: BCH inhibited L-leucine transport in a concentration-dependent manner, and it inhibited cell growth in a time-dependent manner in KB human oral epidermoid carcinoma cells, Saos2 human osteogenic sarcoma cells and C6 rat glioma cells. The formation of a DNA ladder was observed, and the number of TUNEL-positive cells was increased with BCH treatment. Furthermore, the proteolytic processing of caspase-3 in KB and C6 cells and of caspase-7 in KB, Saos2 and C6 cells was increased by BCH treatment. CONCLUSION: These results suggest that the inhibition of LAT1 activity by BCH leads to apoptotic cancer cell death by inducing intracellular depletion of neutral amino acids necessary for cancer cell growth.     

MMP-2、MMP-9在大鼠皮肤创面愈合过程中的表达

目的探讨基质金属蛋白酶2(MMP-2)、MMP-9在大鼠皮肤创面愈合过程中表达的动态及其相关性。方法 42只大鼠制作大鼠背部皮肤创面模型后按取创面皮肤时间随机均分为七组,即在创面形成后1、12、24h及3、7、14、21d切取背部皮肤创面标本,采用免疫组化法检测标本中MMP-2及MMP-9的表达。另取3只正常大鼠背部皮肤标本作为对照。结果正常大鼠皮肤中MMP-2、MMP-9几无表达;背部皮肤创面形成后,MMP-2逐渐升高,14d达峰,21d仍高于正常水平;MMP-9迅速升高,在24h-7d呈高水平表达,后逐渐降至较低水平;MMP-2与MMP-9的表达水平在12h,3、7、14、21d时呈正相关,在24h时呈负相关。结论在大鼠创面愈合过程中MMP-2和MMP-9的表达有一定规律,并且二者相互影响。     

虎杖苷对大鼠皮肤创伤愈合的治疗作用的实验研究

目的 探讨虎杖苷对大鼠皮肤急性创面愈合的治疗作用及可能机制.方法 54只大鼠分为三组:正常组(n=6)、对照组(n=24)及虎杖苷组(n=24).正常组为无处理,建立背部急性全层皮肤缺损模型后虎杖苷组予以虎杖苷20 mg/kg治疗10天,对照组大鼠予以等量生理盐水治疗.统计各组大鼠创面愈合时间、愈合率,HE染色观察创面形态学变化,试剂盒检测创面组织Ⅰ型、Ⅲ型胶原蛋白,TUNEL法检测创面组织细胞凋亡,westem blot法检测创面组织Bax及Bcl-2表达,荧光法检测创面组织caspase-3活性.结果 与对照组比较,虎杖苷组创面愈合率明显提高,愈合时间明显缩短;与对照组比较,虎杖苷组创伤组织中Ⅰ型胶原含量比同期对照组低,而Ⅲ型胶原含量比同期对照组高;与对照组比较,虎杖苷组凋亡细胞数减少,Bax表达降低,Bcl-2表达增加,caspase-3活性减低.结论 虎杖苷治疗可以促进创面愈合,其机制可能与抑制细胞凋亡有关.     

独一味胶囊对大鼠机械性皮肤损伤的促愈合作用

目的研究独一味胶囊对大鼠机械性皮肤损伤的促愈合作用及其可能机制。方法在SD大鼠背部皮肤造成机械性损伤模型,将损伤模型大鼠分为独一味(生药)4、2、1 g.kg-1组、模型组、阳性对照组,给予大鼠ig独一味胶囊药液或阳性药,连续16 d,观察各创面的愈合情况,计算各创面的愈合率,并于造模后第4、7、10、14、17天分别处死大鼠,取创面标本,分别进行病理组织学检查、毛细血管计数和血管内皮生长因子(VEGF)、表皮生长因子(EGF)的免疫组化测定。结果大鼠灌服独一味胶囊后可明显缩短伤口的愈合时间。造模后第4~14天,生药4.0 g.kg-1组大鼠的创口愈合率、创面组织EGF、VEGF的表达明显高于模型组(P<0.05);造模后第4~7天,生药4.0 g.kg-1组大鼠新生毛细血管计数明显多于模型组(P<0.01)。组织形态学评价结果表明:造模后第4~14天,生药4.0 g.kg-1组大鼠的创口修复情况明显优于模型组,在创口修复早期,能加速成纤维细胞的增生,促进新生毛细血管的生成,减少炎症反应。结论独一味胶囊能促进机械性皮肤损伤的愈合,其作用机制可能与促进新生毛细血管生成,促进组织中内源性EGF、VEGF等相关因子的表达有关。     

蜗牛多肽对大鼠皮肤烫伤创面的愈合及碱性成纤维生长因子 mRNA表达的影响

目的：探讨蜗牛多肽（ SP）对大鼠皮肤烫伤的创面愈合及碱性成纤维生长因子（ bFGF） mRNA表达的影响。方法60只大鼠用90℃热水建立深Ⅱ度烫伤模型,建模后随机分为模型组和实验组,实验组大鼠用SP溶液（10 mg? mL－1）涂抹烫伤部位,模型组大鼠皮肤烫伤处涂抹等量0．9％NaCl。观察各组大鼠的皮肤愈合率、肤愈时间及脱痂时间, RT－PCR检测皮肤组织中bFGF mRNA表达。结果实验组的烫伤面积愈合率在用药后不同时段均明显高于模型组（P＜0．05,P＜0．01）;与模型组比较,实验组烫伤皮肤的愈合时间与脱痂时间均明显缩短（P＜0．05）;同时,SP增加了不同时间bFGF mRNA的表达。结论 SP可促进大鼠皮肤烫伤创面愈合,其作用机制可能与调节bFGF mRNA表达有关。     

Inhibitors of mammalian central nervous system selective amino acid transporters

Central nervous system (CNS) selective amino acid transporters provide an important function in maintaining tonic extracellular levels of amino acids that act as neurotransmitters, synaptic modulators or neurotransmitter precursors. Small molecule inhibitors of these transporters have been postulated and in some cases demonstrated to be useful in the treatment of a range of CNS driven disorders such as epilepsy, anxiety, psychosis, depression, pain and neurodegenerative disease. Although much of the research to date in this field has focussed on inhibition of the gama-amino butyric acid (GABA) transporters more recent reports have also generated interest in modulation of glycine, glutamate and proline transporters. This article will review the current medicinal chemistry literature and structure activity relationships known for mammalian CNS selective amino acid transporters.     

创面愈合过程中内源性EGF变化的研究

本文通过免疫组织化学方法,对大鼠创面愈合过程中内源性表皮生长因子(EGF)变化进行了研究,结果表明:创面愈合过程中内源性EGF有规律性变化。对内源性表皮生长因子的作用进行了讨论,创面愈合过程中,结合内源性EGF的变化,合理应用EGF可能会取得更好的效果。     

Application of chitosan-based nanoparticles in skin wound healing

The rising prevalence of impaired wound healing and the consequential healthcare burdens have gained increased attention over recent years. This has prompted research into the development of novel wound dressings with augmented wound healing functions. Nanoparticle(NP)-based delivery systems have become attractive candidates in constructing such wound dressings due to their various favourable attributes. The non-toxicity, biocompatibility and bioactivity of chitosan(CS)-based NPs make them ideal...     

Ethanol extract of Cotinus coggygria leaves accelerates wound healing process in diabetic rats

Context: Cotinus coggygria Scop. (Anacardiaceae) leaves that were used as wound healing in traditional Balkan and Anatolian folk medicine, could be potentially effective in treating diabetic wounds.

Objective: This study investigates biochemical and histological effects of ethanol extract of C. coggygria (CCE) on excision wound model in diabetic rats.

Materials and methods: This study was conducted on diabetic Wistar albino rats, which were injected by a single dose (50?mg/kg i.p.) streptozotocin. Afterward an excision wound model was created in all animals; diabetic control rats were applied topically simple ointment and diabetic treatment rats were applied topically 5% (w/w) ointment with CC, once a day during the experimental period. Malondialdehyde, glutathione and hydroxyproline levels in wound tissues were investigated at the end of 3rd, 7th, and 14th days. Histopathological examination was also performed.

Results: Hydroxyproline content was significantly increased in the CCE treated group versus control after the 3rd and 7th days (15.33 versus 11.83; 19.67 versus 15.67?mg/g, p?<?0.05; respectively). A statistically significant elevation in glutathione at the end of 3rd, 7th, and 14th days (5.13 versus 1.58, p?<?0.05; 4.72 versus 1.88, p?<?0.05; 3.83 versus 1.88?μmol/g, p?<?0.05, respectively) and a statistically significant decrease in malondialdehyde level at the end of 7th day (4.49 versus 1.48?nmol/g, p?<?0.05) were determined in the treated group versus control group. These results were also supported by histological analyses.

Discussion and conclusion: These findings indicate that CCE accelerated the cutaneous wound healing process in diabetic wounds, in confirmation of its traditional use.     

重组人角质细胞生长因子-2涂膜剂促进大鼠皮肤伤口愈合

目的研究重组人角质细胞生长因子-2(Recombinanthumankeratinocytegrowthfactor-2,rhKGF-2)涂膜剂对大鼠皮肤伤口愈合速度的影响。方法(1)用预热100℃的100g砝码在大鼠背部脱毛部位烫8s,形成4个直径约2.5cm的深II度烫伤口;(2)手术方法切除大鼠背部部分皮肤,形成2个1.5cm×4cm的皮肤全层缺损伤口。造模后第2天开始每天局部给药1次,连续15d,每4d测量伤口面积并计算愈合率,第16天取伤口及其周围皮肤组织,石蜡切片,Masson’s三色法染色,MOTICAdvanced3.2形态分析软件分析新生上皮面积、厚度和上皮细胞迁移距离。结果rhKGF-2涂膜剂对大鼠皮肤深II度烫伤和皮肤全层切除伤口的愈合均有明显的促进作用,使新生上皮面积、平均厚度和上皮细胞移行距离等明显增加。作用强度比同剂量的单纯的rhKGF-2溶液剂好。结论rhKGF-2涂膜剂能明显加快大鼠皮肤烫伤和皮肤全层切除伤口周围上皮细胞的增生与迁移,加快创面愈合过程。     

Changes in the biosynthetic activity of gastric glycoproteins during the healing process of acetic acid ulcer in rats

Changes in the incorporation of 3H-glucosamine into the macromolecular glycoprotein during the healing process of acetic acid induced gastric ulcer in rats were sequentially examined in the ulcer region and the intact region at 2, 10, 40, 80 and 365 days after the operation. 1) The total radioactivity (Tissue + Medium) and the radioactivity which remained in the tissue after incubation of the ulcer region were increased significantly as compared with those of the control at 2 days after the operation (275, 175% of the control, respectively), and then total radioactivity returned to the control level. On the other hand, the radioactivity in the tissue was gradually decreased, and then it became 50% of the control at 365 days. In contrast, the incorporating activity into the macromolecular glycoproteins was decreased to 50% of the control at 2 days, and was once recovered to the control level at 10 days. After 40 days, it was again decreased to 50% of the control and became 30% at 365 days. 2) Changes in the incorporation of 3H-glucosamine into the macromolecular glycoproteins of the intact region of rats with ulcers were the same as that of the ulcer region. 3) Elution profiles of gel filtration of the macromolecular glycoproteins isolated from the relapse and recurrence region of rats with ulcers at 365 days were the same as that of the healing region, and their radioactivities were decreased to 30% of the control. The results suggested that such a decrease in the biosynthetic activity of the macromolecular glycoproteins extending over the whole gastric tissue is one of the reasons for the increased relapse and recurrence.     

Electrospun fibers with plasmid bFGF polyplex loadings promote skin wound healing in diabetic rats

Deep or chronic skin wounds are difficult to heal spontaneously due to the lack of scaffold to guide cell growth and reduced levels and activities of endogenous growth factors. Emulsion electrospinning process integrated with DNA condensation techniques indicated potentials to gradually release DNA, but no attempt has been made to clarify the advantages in promoting tissue regeneration and wound recovery. In this study, polyplexes of basic fibroblast growth factor-encoding plasmid (pbFGF) with poly(ethylene imine) were incorporated into electrospun fibers with a core-sheath structure, and poly(ethylene glycol) was included into the fiber sheath to allow a sustained release of pbFGF for 4 weeks. In vitro tests on mouse embryo fibroblasts indicated that pbFGF-loaded fibrous mats enhanced cell proliferation by the autocrine bFGF, and an effective cell transfection proceeded for over 28 days. Skin wounds were created in the dorsal area of diabetic rats for in vivo evaluation of skin regeneration after being covered with pbFGF-loaded fibrous mats. The gradual pbFGF release revealed significantly higher wound recovery rate with improved vascularization, enhanced collagen deposition and maturation, complete re-epithelialization and formation of skin appendages. The above results demonstrate the potential use of pbFGF-loaded electrospun fibrous mats to accelerate the healing of skin ulcers for patients with diabetic mellitus.     

Excitatory amino acid transporters expressed by synovial fibroblasts in rats with collagen-induced arthritis

Although previous studies have demonstrated increased levels of the brain neurotransmitter glutamate (Glu) in the synovial fluid from patients with arthritis, not much attention has been paid to the possible role of Glu in joint synovial tissues to date. Constitutive expression of mRNA was for the first time shown with glutamate aspartate transporter, glutamate transporter-1 and excitatory amino acid carrier-1 (EAAC1), in addition to with particular ionotropic and metabotropic Glu receptors, in cultured synovial fibroblasts prepared from knee joints of male Lewis rats. Immunohistochemical analysis revealed high localization of immunoreactive EAAC1 at synovial tissues. The accumulation of [³H]Glu occurred in a temperature- and sodium-dependent manner in cultured synovial fibroblasts, with a K_m of 23.1 ± 1.1 μM and a V_max of 237.1 ± 31.1 pmol/(mg protein min), respectively. In rats with arthritis induced by immunization to type-II collagen, marked increases were seen in hind paw volume, cytokine mRNA expression and Glu levels in synovial tissues, in addition to histological erosion. In cultured synovial fibroblasts prepared from these arthritic rats, [³H]Glu accumulation was drastically increased with biochemical and pharmacological profiles similar to those seen in normal synovial fibroblasts. The exposure to Glu at 500 μM doubled the incorporation of 5-bromo-2′-deoxyuridine in cultured synovial fibroblasts of arthritic but not normal rats, without significantly affecting mRNA expression of different cytokines in both synovial fibroblasts. These results suggest that Glu may at least in part play a role in mechanisms associated with cellular proliferation through particular transporters functionally expressed by synovium in rheumatoid arthritis.     

Caspase-1在小鼠皮肤切创愈合过程中的规律性表达与损伤时间的关系研究

目的观察小鼠皮肤切创后不同时间内caspase-1的蛋白含量变化,探讨不同时间点蛋白表达的规律与损伤时间的关系。方法建立3个组别。制作小鼠皮肤切创模型,应用免疫组织化学技术检测切创后不同时间皮肤中caspase-1的表达,同时设立空白对照组和死后切创组。结果空白对照组,死后损伤组及生前损伤组0h的蛋白只表达在表皮层,毛囊,皮脂腺中。伤后3h,切创组及切创周边区可见多核粒细胞浸润且少量多核粒细胞表达caspase-1。6h～1d,部分浸润的多核粒细胞和单核细胞caspase-1阳性。伤后3～5d,成纤维细胞大量增生,此时caspase-1主要表现在单核细胞和成纤维细胞。伤后7d则蛋白表达阳性细胞以成纤维细胞为主。结论小鼠切创愈合过程中caspase-1主要在多核粒细胞、单核细胞及成纤维细胞表达,并随损伤时间延长,呈现规律变化趋势。其时序性变化可用于皮肤切创损伤时间的推断。     

Pharmaceutical application of frog skin on full-thickness skin wound healing in mice

Abstract

Context: It has been proved that fresh frog skin is efficient in the wound healing process.

Objective: The purpose of study is to introduce a formulation of frog skin powder for evaluation of wound repair where fresh frog skin is not available.

Materials and methods: Rana ridibunda (Ranidae) skins were lyophilized, and a powder was prepared. The powder (0.0005?g) was then mixed with ointment (0.0065?g) for treating each wound. Formulation was used on full-thickness wounds on mice (FO group) and compared to positive and negative controls. In order to study the wound healing process, wound contraction, inflammation, number of fibroblast cells, neovascularization and collagen density were evaluated on days 2, 4 and 6 following the injury. Moreover, CFU measurement was performed for the evaluation of wound contamination.

Results: Acceleration in wound contraction in the FO group compared to control groups was significant (p?<?0.001) on days 4 and 6. Results showed that FO treatment considerably decreased inflammatory cells during the study. On day 4, FO treatment was significantly effective in increasing the number of fibroblast cells and collagen density (p?<?0.01 and p?<?0.05, respectively). On day 6 the number of fibroblast cells (p?<?0.001), collagen density (p?<?0.05) and neovascularization (p?<?0.05), were higher in the FO group than the control groups. Results of CFU measurement demonstrated significant reduction of wound contamination in FO treated wounds on days 2 (p?<?0.05) and 4 (p?<?0.01).

Discussion and conclusion: Our findings indicated that the pharmaceutical form of frog skin used in this study has considerable healing and antibacterial effects on wounds.     

Effect of cimetidine on wound healing in rats

Cimetidine, an H-2 antagonist, is one of the most commonly prescribed drugs and is used in the management of peptic ulcer disease and prevention of stress ulcerations following major trauma or operations. It has been shown that histamine has a stimulatory effect on wound healing which is believed to be mediated through H-2 receptors. We hypothesized that cimetidine would have an inhibitory effect on wound healing. Fifty-two adult rats were divided into two groups: Group A was placed on a standard chow and Group B was given chow supplemented with cimetidine (300 mg/Kg diet), a dose equivalent to that clinically used in man and effective in inhibiting gastric hypersecretion in the rat. Ten days later, all rats underwent a dorsal skin incision under pentobarbital anesthesia and implantation of a polyvinyl alcohol sponge subcutaneously. Animals were maintained on their respective diets. Rats were killed with ether five and ten days postoperatively, the wounds were excised and breaking strength measured. Sponge hydroxyproline content was measured. No significant difference in breaking strength or sponge hydroxyproline content was found between the two groups. We thus conclude that cimetidine has no effect on wound healing in rats when used in doses equivalent to the clinical dose in man.