Characterization of an arabinogalactan protein from the pressed juice of <Emphasis Type="Italic">Echinacea purpurea</Emphasis>: investigations into the type of linkage between the protein and polysaccharide moieties

A high molecular weight arabinogalactan protein (AGP) from the pressed juice of Echinacea purpurea, known to exhibit immunomodulatory properties in vitro, was characterized. Alkaline hydrolysis was carried out, leading to degradation of the protein core and to carbohydrate moieties linked to the amino acid responsible for binding. Gel permeation chromatography of these AG subunits gave one peak with a molecular weight of 30 × 10³ Da. Hydroxyproline (42.9% w/w) was detected as the dominant amino acid after alkaline hydrolysis and was thus identified as the major amino acid responsible for the binding between the protein and the AG subunits via an O-glycosidic linkage. Large amounts of glutamine/glutamic acid (24.5% w/w) and asparagine/aspartic acid (17.3% w/w) were also found. Polyclonal antibodies raised against the intact AGP were shown to bind to the AG subunits too, indicating that the epitopes responsible for such interactions are localized in the polysaccharide moiety of the AGP.     

Laser photobioactivation mechanisms: in vitro studies using ascorbic acid uptake and hydroxyproline formation as biochemical markers of irradiation response

Clinical investigations of laser photobioactivation, or biostimulation, might be differently designed and more fruitful if knowledge of basic biochemical mechanisms were better understood. In this investigation, biochemical events identified as responses to 904 nm irradiation included increased ascorbic acid uptake by fibroblasts. These cells also showed increased hydroxyproline formation, and this was increased several-fold by the addition of proline to the medium. Maximum biochemical responses were observed at a pulse frequency of 67 Hz and a pulse width of 150 nsec with an energy density of approximately 7 mJ/cm2 per exposure. Elements in the mitochondrial cytochrome system are proposed as the radiation absorbing chromophore(s). Hypothetically, the energy generated is linked to ascorbic acid uptake, which in turn stimulates collagen synthesis.     

动物组织中羟脯氨酸测定方法的建立及初步应用

目的：建立动物组织中的羟脯氨酸测定方法,以之估计胶原含量并衡量组织愈合程度。方法：研究显色温度、氧化时间、样品中盐浓度以及对 二甲氨基苯甲醛（ Ｐ Ｄ Ａ Ｂ） 浓度对羟脯氨酸测定的影响,据此建立动物组织中的羟脯氨酸测定方法,并初步应用于实验动物犬贲门胶原含量的测定。结果：根据优化的实验结果,羟脯氨酸测定的条件为室温氧化４ ｍｉｎ ,１０ ％ Ｐ Ｄ Ａ Ｂ,１００ ℃显色２ ｍｉｎ 。该法回收率为９８ ％ ,变异系数为４ ％ 。结论：该法重复性好,可信度高,可作为临床判定愈合程度的一个定量指标。     

雌孕激素合用对去卵巢大鼠骨质疏松的预防作用

目的　观察炔诺酮和炔雌醇联合应用对去卵巢大鼠骨质疏松的预防作用。方法　 4个半月龄大鼠双侧卵巢去除术后预防用药 90d。用骨组织形态计量学方法 ,测定大鼠胫骨近端松质骨静态参数和动态参数 ,观察骨和尿中有机质和无机质的含量 ,血清生化指标和器官指数。结果　炔诺酮6 0 μg·kg-1·d-1和炔雌醇 3 5 μg·kg-1·d-1联合应用可抑制去卵巢大鼠的破骨细胞活性 ,完全对抗其骨转化率增高和骨量丢失的骨质疏松症状 ,同时可使去卵巢大鼠的骨有机质含量增加和尿羟脯氨酸排出量减少。结论　炔诺酮和炔雌醇联合应用可预防去卵巢大鼠骨质疏松     

木犀草素对博莱霉素引起的大鼠肺纤维化模型的治疗作用

目的:研究木犀草素对大鼠肺纤维化的治疗作用。方法:观察不同剂量的木犀草素对博莱霉素诱导的大鼠肺纤维化肺组织的肺重量指数、丙二醛(MDA)、羟脯氨酸(HYP)含量及组织学变化的影响。结果:木犀草素在20,40和80mg·kg~(-1)剂量下连续灌胃给药21d,对模型大鼠的上述指标有不同程度的抑制作用,对其组织形态也有明显改善,且呈剂量效应关系。结论:木犀草素对大鼠肺纤维化模型具有较好的治疗效果。     

Expression of muscarinic acetylcholine receptors in hepatocytes from rat fibrotic liver

The pathological changes of parasympathetic nerve are considered as an independent prognostic factor of the survival rate of patients with chronic liver disease. The non-selective muscarinic acetylcholine receptors (mAchR) agonists and antagonists can affect the proliferation of hepatocytes and hepatic stellate cells, but the subtypes of mAchR expressions in HCs are still uncertain. Here, we investigate the expression of mAchR in hepatic fibrosis on rats. 3 ml/kg 40% carbon tetrachloride (CCL4) was given to induce hepatic fibrosis on rats and the hepatocytes were isolated. Compared to the normal state, the expression levels of m1, 3, 5 in fibrotic liver tissues or hepatocytes were obviously increased, while m2, 4 decreased. 10 μM pilocarpine or 10 μM acetylcholine could increase the alanine aminotransferase (ALT), hydroxyproline (Hyp), collagen I, III in the hepatocytes, and decreased albumin (ALB). They also changed the expressions of mAchR similarly as the fibrotic hepatocytes and livers. However, atropine could ameliorate the state of fibrotic hepatocytes. These data indicate that mAchR played an important role in the regulation of hepatic fibrosis process. Targeting mAchR would have therapeutic potential for hepatic fibrosis.     

Melatonin reduces dimethylnitrosamine-induced liver fibrosis in rats

Increased deposition of the extracellular matrix components, particularly collagen, is a central phenomenon in liver fibrosis. Stellate cells, the central mediators in the pathogenesis of fibrosis are activated by free radicals, and synthesize collagen. Melatonin is a potent physiological scavenger of hydroxyl radicals. Melatonin has also been shown to be involved in the inhibitory regulation of collagen content in tissues. At present, no effective treatment of liver fibrosis is available for clinical use. We aimed to test the effects of melatonin on dimethylnitrosamine (DMN)-induced liver damage in rats. Wistar albino rats were injected with DMN intraperitoneally. Following a single dose of 40 mg/kg DMN, either saline (DMN) or 100 mg/kg daily melatonin was administered for 14 days. In other rats, physiologic saline or melatonin were injected for 14 days, following a single injection of saline as control. Hepatic fibrotic changes were evaluated biochemically by measuring tissue hydroxyproline levels and histopathogical examination. Malondialdehyde (MDA), an end product of lipid peroxidation, and glutathione (GSH) and superoxide dismutase (SOD) levels were evaluated in blood and tissue homogenates. DMN caused hepatic fibrotic changes, whereas melatonin suppressed these changes in five of 14 rats (P < 0.05). DMN administration resulted in increased hydroxyproline and MDA levels, and decreased GSH and SOD levels, whereas melatonin reversed these effects. When melatonin was administered alone, no significant changes in biochemical parameters were noted. In conclusion, the present study suggests that melatonin functions as a potent fibrosuppressant and antioxidant, and may be a therapeutic choice.     

Effect of Calendula Officinalis Cream on Achilles Tendon Healing

In recent years, the scientific community has undertaken research on plant extracts, searching for compounds with pharmacological activities that can be used in diverse fields of medicine. Calendula officinalis L. is known to have antioxidant, anti‐inflammatory, antibacterial, and wound healing properties when used to treat skin burns. Therefore, the purpose of this study was to analyze the effects of C. officinalis on the initial phase of Achilles tendon healing. Wistar rats were separated in three groups: Calendula (Cal)—rats with a transected tendon were treated with topical applications of C. officinalis cream and then euthanized 7 days after injury; Control (C)—rats were treated with only vehicle after transection; and Normal (N)—rats without tenotomy. Higher concentrations of hydroxyproline (an indicator of total collagen) and non‐collagenous proteins were observed in the Cal group in relation to the C group. Zymography showed no difference in the amount of the isoforms of metalloproteinase‐2 and of metalloproteinase‐9, between C and Cal groups. Polarization microscopy images analysis showed that the Cal group presented a slightly higher birefringence compared with the C group. In sections of tendons stained with toluidine blue, the transected groups presented higher metachromasy as compared with the N group. Immunocytochemistry analysis for chondroitin‐6‐sulfate showed no difference between the C and Cal groups. In conclusion, the topical application of C. officinalis after tendon transection increases the concentrations of collagen and non‐collagenous proteins, as well as the collagen organization in the initial phase of healing. Anat Rec, 298:428–435, 2015. © 2014 Wiley Periodicals, Inc.     

Effects of Neurotensin on the Healing of Experimental Anastomosis of the Colon

The leakage of colonic anastomoses increases perioperative morbidity significantly. The purpose of the study was to investigate the influence of neurotensin, an intestinal trophic peptide, on the healing of colonic anastomosis. Forty-two Wistar-albino rats were separated into three equal groups: Group 1 (control group) — segmental resection of the left colon and end-to-end anastomosis; Group 2 (dexamethasone group) — resection and anastomosis, plus 0.1 mg/kg/day of dexamethasone; Group 3 (neurotensin group) — same surgical procedure plus 300 pg/kg/day of neurotensin. Bursting pressure and tissue hydroxyproline content were determined as parameters of the anastomosis strength and healing on the 3^rd and 7^th days postoperatively. On the 3^rd day, mean bursting pressures were 141.4, 146.7 and 73.1 (p = 0.0001) cm of water in the control group, dexamethasone and neurotensin groups respectively. On the 7^th day, bursting pressures were measured as 237.4, 100.6 (p = 0.0001) and 72.7(p = 10^-6) cm of water, in the control group, dexamethasone and neurotensin groups respectively. Between the 3^rd and 7^th days, bursting pressures were increased significantly in the control group (p = 0.0001), decreased in the dexamethasone (p = 0.048), and maintained their lowest values in the neurotensin (p = 0.96) groups. On the 7^th day, mean hydroxyproline levels were measured as 9.20, 3.30 (p = 0.007), 2.86 (p = 0.007) pg, in the control group, dexamethasone, and neurotensin groups respectively. Between the 3^rd and 7^th days, tissue hydroxyproline levels were increased significantly in the control group (p = 0.004), decreased in the dexamethasone (p = 0.03), and maintained their lowest values in the neurotensin (p = 0.87) groups. The anastomosis resistance to intraluminal pressure was weak, tissue collagen content was insufficient, and healing was inadequate in the dexamethasone and neurotensin groups in respect to the control group. We concluded that neurotensin impaired the healing, and affected the strength of the colonic anastomosis.     

Resterilized Mesh in Repair of Abdominal Wall Defects in Rats

A variety of negative opinions about repeated usage of relatively expensive resterilized synthetic meshes have been considered. It had been stated that resterilized polypropylene meshes inhibits fibroblastic activity, decreases proliferative activity, and increases apoptosis in human fibroblast culture, in vitro. The purpose of this study is the in vivo evaluation of the resterilized mesh repairs of abdominal hernia defects in rat models of incisional hernia by comparing primer repair and original mesh repairs. The rats (n = 22) were separated into three groups. While the abdominal defect was repaired by primary suture in the control group (CG), the defects were repaired by original mesh (OG) or resterilized mesh (RG) in mesh-repaired groups. After 21 days, the rats were evaluated for tissue tensile strengths, tissue hydroxyproline levels, tissue inflammation, fibrosis, and apoptosis. Although the tensile strengths in OG and RG were significantly higher than those of CG (p <. 05 and p <. 05), there was no significant difference between two groups. The tissue hydroxyproline levels in OG and RG were also higher than those of CG. The difference was not significant between the two groups. The inflammation and fibrosis indexes in OG and RG were significantly higher than those of CG (p <. 0001 for both), but there was no difference between groups. While the apoptosis index in OG and RG was also higher than that of CG (p <. 0001 for both), there was no significant difference between OG and RG. The usage of resterilized mesh in abdominal wall repair did not reduce the tissue tensile strength, did not affect the tissue hydroxyproline levels, did not decrease the fibrosis, and did not increase the tissue inflammation and apoptosis. In conclusion, usage of resterilized meshes in abdominal wall defects was as safe as sterilized meshes.