苦黄注射液联合复方蛋氨酸胆碱治疗酒精性脂肪肝的疗效及对肝纤维化的影响研究

目的 探究复方蛋氨酸胆碱结合苦黄注射液在酒精性脂肪肝治疗中的疗效及对肝纤维化的影响。方法 按照随机数字表法将2019年1月至2021年12月海安市人民医院收治并确诊的104例酒精性脂肪肝患者分为对照组和观察组,各52例。对照组给予复方蛋氨酸胆碱治疗,观察组苦黄注射液联合复方蛋氨酸胆碱治疗。两组患者均连续治疗1个月。采用全自动生化分析仪检测两组患者治疗前后血清丙氨酸转移酶（alanine transferase,ALT）、天冬氨酸转氨酶（aspartate aminotransferase,AST）、总胆固醇（total cholesterol ,TC）、三酰甘油（triacylglycerol,TG）、总胆红素（total bilirubin,TBil）水平;采用放射免疫法检测两组患者治疗前后血清层粘连蛋白（laminin,LN）、透明质酸（hyaluronic acid,HA）、Ⅲ型前胶原N端肽（type III procollagen N-terminal peptide,PC Ⅲ）及Ⅳ型胶原（type IV collagen,Ⅳ-C）水平。比较两组患者的临床症状改善情况、临床疗效、肝功能指标及肝纤维化指标。结果 观察组治疗后的腹胀、肝区不适、黄疸、痞满等症状消失时间分别为（3.9±0.1） d、（9.8±0.2） d、（11.5±0.7）d、（3.8±0.2） d,均短于对照组分别为（7.5±0.3） d、（14.3±0.4） d、（15.6±0.8） d、（7.4±0.3） d（均P<0.05）。观察组临床总有效率高于对照组（94.2% vs 78.9%）（χ2=5.283,P=0.022）。观察组ALT、AST、TC、TG、TBil水平分别为（31.5±5.3） IU/L、（36.4±3.4） IU/L、（4.1±0.2） mmol/L、（1.4±0.3） mmol/L、（12.6±3.1） μmol/L,低于对照组[分别为（57.4±5.6） IU/L、（54.7±3.5） IU/L、（6.4±0.4） mmol/L、（2.6±0.5） mmol/L、（21.2±5.3） μmol/L]（均P<0.05）。观察组治疗后LN、HA、PC Ⅲ、Ⅳ-C水平分别为（51.6±9.4） μg/L、（32.5±7.4） μg/L、（7.5±1.8） ng/mL、（32.3±4.2） μg/L,均低于对照组[分别为（80.5±10.3） μg/L、（58.7±12.2） μg/L、（16.5±3.6） ng/mL、（71.2±8.6） μg/L]（均P<0.05）。对照组和观察组不良反应发生率分别为7.68%、3.84%,两组比较无明显差异（χ2=0.707,P=0.400）。结论 复方蛋氨酸胆碱结合苦黄注射液在酒精性脂肪肝患者中的治疗疗效显著,可改善患者的临床症状和肝功能,延缓肝纤维化的进程,安全性高。     

Does steatohepatitis impair liver regeneration? A study in a dietary model of non-alcoholic steatohepatitis in rats

BACKGROUND: Impaired liver regeneration is a feature of alcoholic hepatitis, but the relative importance of alcohol, nutritional imbalance and inflammatory mediators in causing this effect is unclear. Non-alcoholic steatohepatitis (NASH) is a form of liver disease with similar morphology to alcoholic hepatitis, but the effect of this disorder on liver regeneration is unclear. We, therefore, examined the status of liver regeneration in a rat nutritional model of hepatic steatosis with inflammation, which is morphologically identical to NASH in humans. Methods: Male Wistar rats received a methionine-choline-deficient diet (MCDD) for 4 weeks before experiments and both isocaloric pair-fed and ad libitum-fed rats were used as controls. Following partial hepatectomy (68%), the extent of hepatic regeneration was determined 24 h later using [3H]-thymidine incorporation and restitution of liver mass. Results: There was no significant difference of [3H]-thymidine incorporation in MCDD-fed, pair-fed and ad libitum-fed rats (80+/-27, 78+/-11 and 80+/-6.3 d.p.m./microg DNA, respectively). Similarly, restituted liver masses in three groups of rats were not significantly different (17+/-3.8, 18+/-1.8 and 17+/-3.1% initial liver weight, respectively). Conclusions: The similarities in hepatic histology and cytochrome P450 2E1 induction between this nutritional model of hepatic steatohepatitis and alcoholic steatohepatitis imply that these two disorders share pathogenetic mechanisms. However, liver regeneration is not altered by NASH in rats, indicating that the nutritional and inflammatory changes that appear similar to those of alcoholic liver disease do not cause impairment of liver regeneration.     

Distinct Localization of Peripheral and Central Types of Choline Acetyltransferase in the Rat Cochlea

We previously discovered a splice variant of choline acetyltransferase (ChAT) mRNA, and designated the variant protein pChAT because of its preferential expression in peripheral neuronal structures. In this study, we examined the immunohistochemical localization of pChAT in rat cochlea and compared the distribution pattern to those of common ChAT (cChAT) and acetylcholinesterase. Some neuronal cell bodies and fibers in the spiral ganglia showed immunoreactivity for pChAT, predominantly the small spiral ganglion cells, indicating outer hair cell type II neurons. In contrast, cChAT- and acetylcholinesterase-positive structures were localized to fibers and not apparent in ganglion cells. After ablation of the cochlear nuclei, many pChAT-positive cochlear nerve fibers became clearly visible, whereas fibers immunopositive for cChAT and acetylcholine esterase disappeared. These results suggested that pChAT and cChAT are localized in different systems of the rat cochlea; pChAT in the afferent and cChAT in the efferent structures.     

Distribution and morphology of sensory and autonomic fibres in the subendocardial plexus of the rat heart

Cardiac reflexes originating from sensory receptors in the heart ensure blood supply to vital tissues and organs in the face of constantly changing demands. Atrial volume receptors are mechanically sensitive vagal afferents which relay to the medulla and hypothalamus, affecting vasopressin release and renal sympathetic activity. To date, two anatomically distinct sensory endings have been identified which may subserve cardiac mechanosensation: end-nets and flower-spray endings. To map the distribution of atrial receptors in the subendocardial space, we have double-labelled rat right atrial whole mounts for neurofilament heavy chain (NFH) and synaptic vesicle protein 2 (SV2) and generated high-resolution maps of the rat subendocardial neural plexus at the cavo-atrial region. In order to elucidate the nature of these fibres, double labelling with synaptophysin (SYN) and either NFH, calcitonin gene-related peptide (CGRP), choline acetyltransferase (ChAT) or tyrosine hydroxylase (TH) was performed. The findings show that subendocardial nerve nets are denser at the superior cavo-atrial junction than the mid-atrial region. Adluminal plexuses had the finest diameters and stained positively for synaptic vesicles (SV2 and SYN), CGRP and TH. These plexuses may represent sympathetic post-ganglionic fibres and/or sensory afferents. The latter are candidate substrates for type B volume receptors which are excited by stretch during atrial filling. Deeper nerve fibres appeared coarser and may be cholinergic (positive staining for ChAT). Flower-spray endings were never observed using immunohistochemistry but were delineated clearly with the intravital stain methylene blue. We suggest that differing nerve fibre structures form the basis by which atrial deformation and hence atrial filling is reflected to the brain.     

Radioactive choline metabolism in guinea pig gallbladder

Acetylcholine may be released from gallbladder intrinsic nerves in response to cholecystokinin stimulation. This study characterized metabolites of [¹⁴C]choline produced in the gallbladder and released during incubation, with or without cholecystokinin-octapeptide. Radiolabeled [¹⁴C]choline was applied to the mucosal or muscle surface of intact guinea pig gallbladders in an organ bath. After radiolabeling, gallbladders were incubated with or without the contractile agonist cholecystokinin-octapeptide. Metabolites of [¹⁴C]choline were identified in gallbladder tissue and incubation buffers using HPLC and thin-layer chromatography. The major metabolites of [¹⁴C]choline were betaine and phosphocholine. [¹⁴C]Phosphocholine was incorporated slowly into [¹⁴C]phosphatidylcholine. [¹⁴C]Choline was released into buffers during incubation. [¹⁴C]Acetylcholine constituted less than 1% of radiolabel in the gallbladder. There was no identifiable [¹⁴C]acetylcholine released in buffers. Cholecystokinin-octapeptide did not affect choline metabolism. These studies showed that choline in the gallbladder is metabolized along pathways similar to those in the liver. Gallbladders released mostly choline, rather than acetylcholine, even during hormonally induced contraction.This project was supported by the Research and Development Office of the Department of Veterans Affairs.     

Focal differences in lipid metabolism of the young pig aorta : Part 2. Synthesis from [2-C]ethanolamine and [1,2-C]choline

The ability of normal young pig aortic tissue to synthesize phospholipids from [2-¹⁴C]ethanolamine and [1,2-¹⁴C]choline, in vitro, has been examined in areas of focal Evans Blue accumulation (blue areas) and adjacent areas of no dye accumulation (white areas).

Incorporation of [2-¹⁴C]ethanolamine into total lipid was linear for 3 h of incubation in both blue and white areas. At 3 h, ethanolamine incorporation into phosphatidyl ethanolamine was significantly less in blue than in white areas.

[1,2-¹⁴C]Choline incorporation into total lipid was linear for 3 h of incubation in blue areas but not in white areas. At 30 min, choline incorporation into phosphatidyl choline was significantly less in blue than in white areas; at 1 h choline incorporation into phosphatidyl choline was similar in blue and white areas, while after 3 h of incubation incorporation was significantly greater in blue than in white areas.

With both [2-¹⁴C]ethanolamine and [1,2-¹⁴C]choline, the percentage distribution of label among individual phospholipids was similar in blue and white areas.

Phospholipid content of blue and white areas was similar.

The results presented demonstrate further focal metabolic differences within the same geographical region of the normal young pig aorta.     

Intestinal microbial metabolism of phosphatidylcholine: a novel insight in the cardiovascular risk scenario

Intestinal microbiota is a “dynamic organ” influencing host metabolism, nutrition, physiology and immune system. Among its several interactions, the role of a phosphatidylcholine metabolite derived by gut flora activity, i.e., trimethylamine-N-oxide (TMAO), allows perceiving a novel insight in the cardiovascular risk scenario, being a strong predictor of this condition. Based on current reports, including the paper of Tang et al., we describe here: the possible role of intestinal microbiota in cardiovascular risk as well as potential interventions to reduce gut flora TMAO production by diet, probiotics and antibiotics. Finally, we highlight the possibility of evaluating, monitoring and modulating TMAO in order to use its serum levels as a marker of cardiovascular risk in the next future, when the need of controlled studies on large series will be satisfied.     

A C-terminal truncated mutation of licC attenuates the virulence of Streptococcus pneumoniae

LicC has been identified as a virulence factor of Streptococcus pneumoniae. However, its role in virulence is still not fully understood because deletion of licC is lethal for the bacterium. In this study, a mutant with 78-bp truncation at the C-terminus of licC was obtained from a signature-tagged mutagenesis (STM) library. The mutant was viable with a large reduction in enzymatic activity as CTP:phosphocholine cytidylyltransferase detected in vitro using a firefly luciferase assay. The mutation attenuated the adhesion and invasion of S. pneumoniae ST556 (serotype 19F) to epithelial cells by 72% and 80%, respectively, and increased the phagocytosis by macrophages for 16.5%, compared to the parental strain. When the mutation was introduced into the encapsulated D39 strain (serotype 2), it led to attenuated virulence in mouse models either by intranasal colonization or by intraperitoneal infection. In addition, the phosphocholine (PCho) on cell surface was decreased, and the choline binding proteins (CBPs) were impaired, which may explain the attenuated virulence of the mutant. These observations indicate that C-terminus of licC is accounted for the main activity of LicC in PCho metabolism and is essential for the virulence of S. pneumoniae, which provides a novel target for drug design against pneumococcal infection.     

Imaging Neuroinflammation? A Perspective from MR Spectroscopy

Neuroinflammatory mechanisms contribute to the brain pathology resulting from human immunodeficiency virus (HIV) infection. Magnetic resonance spectroscopy (MRS) has been touted as a suitable method for discriminating in vivo markers of neuroinflammation. The present MRS study was conducted in four groups: alcohol dependent (A, n = 37), HIV‐infected (H, n = 33), alcohol dependent + HIV infected (HA, n = 38) and healthy control (C, n = 62) individuals to determine whether metabolites would change in a pattern reflecting neuroinflammation. Significant four‐group comparisons were evident only for striatal choline‐containing compounds (Cho) and myo‐inositol (mI), which follow‐up analysis demonstrated were due to higher levels in HA compared with C individuals. To explore the potential relevance of elevated Cho and mI, correlations between blood markers, medication status and alcohol consumption were evaluated in H + HA subjects. Having an acquired immune deficiency syndrome (AIDS)‐defining event or hepatitis C was associated with higher Cho; lower Cho levels, however, were associated with low thiamine levels and with highly active antiretroviral HIV treatment (HAART). Higher levels of mI were related to greater lifetime alcohol consumed, whereas HAART was associated with lower mI levels. The current results suggest that competing mechanisms can influence in vivo Cho and mI levels, and that elevations in these metabolites cannot necessarily be interpreted as reflecting a single underlying mechanism, including neuroinflammation.