脾肾-肌骨-线粒体理论探析

黄宏兴教授课题组长期致力于研究骨质疏松症的中医药防治，首次提出脾肾-肌骨-线粒体理论，历年来的相关研究使得该理论逐步完善。本文通过整理相关资料，明确脾肾、肌骨、线粒体三者关系，精炼出该理论的核心内容：骨质疏松与脾肾、线粒体、肌骨关系密切；脾肾-肌骨-线粒体理论基本内容包含了脾肾共司肌骨、线粒体功能与脾肾相通两个核心内容。脾肾-肌骨-线粒体理论是结合中医学中的“脾肾相关”“脾主肌肉”“肾主骨”和现代生物学中的线粒体功能发展而来，其充分运用中医辨证施治原则，在防治骨质疏松症实际应用中发挥着重要作用。本文通过介绍该理论的演变、发展沿革及应用，为骨质疏松症的防治提供实验研究和临床应用相关材料。     

软骨细胞线粒体氧化应激在骨关节炎发病机制中的研究进展

骨关节炎（OA）是老年人最常见的关节疾病之一,同时也是导致残疾的重要因素。软骨细胞是成熟软骨中唯一的细胞,软骨细胞的稳定和正常功能的维持对延缓OA进展有重要作用。近年来的研究发现,软骨细胞线粒体氧化应激与OA关系密切。本文综述了软骨细胞线粒体氧化应激在OA发病过程中的相关机制,为OA的相关研究提供一定参考。     

肾缺血再灌注损伤与线粒体功能障碍的研究进展

肾缺血再灌注损伤(RIRI)是一个多途径的病理和生理过程,涉及复杂的发生发展机制,其中包含了肾脏线粒体稳态平衡引起的功能紊乱,与肾脏线粒体氧化应激、自噬、凋亡、动力学和再生等密切相关。这些事件相互作用,从多条途径参与了RIRI过程,是其发生发展的重要原因。本文就线粒体与RIRI相关机制的最新研究进展进行综述,为基于线粒...     

基于“肾痰”认识骨-脂代谢异常失衡在绝经后骨质疏松症发病中的作用

绝经后骨质疏松症（PMOP）是绝经后妇女常发生的一种全身性代谢疾病,主要表现为骨量低下、骨微结构退行性改变、骨脆性增加且极易发生脆性骨折,严重威胁着绝经后妇女的健康。现代研究表明绝经后骨代谢紊乱与脂代谢失衡常常并见,中医学则认为PMOP的发病多从肾论治。课题组长期从事PMOP的临床与基础研究工作,认为骨-脂代谢失衡是PMOP的重要发病机制,“肾痰”亦是PMOP发病的关键所在,临床论治除了补肾,还应化痰,并通过一系列实验研究证实了从“肾痰”论治PMOP的可能性。因此,本文拟基于“肾痰”探讨骨-脂代谢失衡在绝经后骨质疏松症发病中的作用,丰富中医药论治PMOP的科学内涵。     

“脾主运化”探讨线粒体铁代谢与骨骼健康的关系

脾居中央,灌四旁,主运化、在体合肉。脾气健运,气血生化有源,脾的合理调节使饮食水谷正常转化为精微物质,经心肺转输和营养全身组织器官,维持全身脏腑组织器官的正常代谢。当脾气虚损时,脾失健运,运化失司,对饮食物的消化功能减弱,水谷精微的吸收不足,则机体代谢异常。线粒体参与合成ATP为细胞提供能量、负责氧化的途径三羧酸循环、氧化磷酸化等功能与气血生化源的“脾脏”功能相类似。线粒体铁代谢失常与很多慢性病有关,如骨质疏松、慢性病贫血、重症肌无力等。线粒体是细胞内铁的代谢场所,铁是一种多功能的辅助因子,对许多代谢途径至关重要。线粒体铁稳态是机体代谢调节中可维持骨骼健康的基础,线粒体铁代谢的稳定调节则促进破骨细胞的正常分化,在骨骼的生长、修复以及重建中起重要作用,而铁超负荷可增加骨骼吸收以及氧化应激,使骨结构和组分发生改变,影响骨骼的健康,从而导致骨质疏松等。因此,本研究基于脾与线粒体铁代谢相关性,从“脾主运化”探讨线粒体铁代谢对骨骼健康的影响及机制。     

从“治痿独取阳明”论治脾肾两虚型老年性骨质疏松症

老年性骨质疏松症(senile osteoporosis,SOP)是一种以骨量减少和骨微结构破坏为特征的老年全身退化性骨病,以致骨的强度降低,骨脆性增加,骨折发生风险较高。SOP属于祖国医学中\"骨痿\"、\"痿证\"、\"骨枯\"等范畴,该病以肝、脾、肾三脏虚损为本,脾肾两虚证是SOP较为常见的证型。目前关于脾肾两虚型SOP是以肾虚为主要病机,补肾健骨成为主要治法,而从脾胃方面论治本病的相关研究较少。本文首次从\"治痿独取阳明\"角度论治脾肾两虚型SOP,补益中焦脾胃以益肾,并滋养脏腑,改善机体营养状况,为防治本病提供新的思路。     

“肾为封藏之本”理论与原发性骨质疏松症的发病机制的研究

原发性骨质疏松症是一种中老年人常见的代谢性骨疾病,根据临床症状属中医“骨痿”范畴,中医药在临床防治中的独特优势使得近年来对本病的中医药和基础理论的研究增多。现代研究中发现一些人体基础物质的功能和特性与中医理论中的肾精功能、作用相似,这些人体基础物质的缺乏是导致本病的主要原因。精亏髓减,骨失所养是本病的核心病机,封藏功能作为肾的主要生理功能,体现在对脏腑精气的贮藏与调控,故原发性骨质疏松症产生、发展多与肾的封藏功能失司相关。本文就《内经》中“肾者,主蛰封藏之本”理论与原发性骨质疏松症的发病机制关系的现代研究,为原发性骨质疏松症的临床防治提供参考。     

基于钙沉积异常探讨原发性骨质疏松症“痰邪”的理论研究

骨质疏松症是以骨强度下降、骨折风险性增加为特征的骨骼系统疾病。与骨骼强度相关的因素主要包括骨矿密度和骨质量,骨矿化是骨代谢的重要过程,而钙的异常沉积是骨矿化异常的表现,是骨质疏松症发病及加重的因素之一。多种骨基质蛋白参与血管钙化等异位钙化的过程,研究发现治疗骨质疏松症的药物也能同时改善血管钙化,因此骨质疏松症的治疗可以从调节钙的沉积入手,通过对骨钙相关蛋白的调节,促进正常的骨矿化而抑制异位钙化,从而达到治疗骨质疏松症的目的。向楠教授领导的课题组在前期的研究中,基于对脂代谢紊乱与骨质疏松症的相关性的认识及痰浊在骨质疏松症发病中作用的认识,提出了"脂代谢异常可能与骨质疏松症痰浊有关"的假说,并制定了补肾化痰的新治则,进行了一系列的实验研究,表明了从"痰"论治骨质疏松症的可行性。现在,根据正常的钙代谢在人体内环境的调节作用及钙的异常沉积引起的病理表现,认为钙的异常沉积亦属于中医"痰邪"的范畴,因此从"痰"论治骨质疏松症还可从调节钙沉积的角度入手,运用补肾化痰法调节钙的沉积,从而达到治疗骨质疏松症的目的,这还有待进一步的研究与证实。     

从肝脾肾探讨原发性骨质疏松症与老年衰弱综合征

骨质疏松症(osteoporosis, OP)是一种全身代谢性骨病,随着我国人口老龄化进程的发展,原发性OP和老年衰弱综合征成为目前常见的两种老年疾病。在中医理论方面,原发性OP以肾虚为本,脾虚为源,肝虚为关键,衰弱多由于老年人五脏俱虚,脾肾尤甚,二者有部分相同的病机。从分子生物学角度,二者在钙磷调节激素、外泌体、营养物质、炎症因子等方面均有联系。通过对相关文献的梳理和总结,从肝脾肾和分子生物学角度寻找原发性OP和衰弱之间的相同点,为后续两种疾病的预防和治疗提供理论依据。     

从“肾-骨-髓-脑”理论探讨外泌体与原发性骨质疏松症的关系

原发性骨质疏松症（primary osteoporosis,POP）是我国亟待解决的公共健康问题。“肾-骨-髓-脑”系统是中医理论体系的重要组成部分,肾精充盛,则髓化生充足,脑得髓充则髓海清明,骨得髓养而健壮有力,构成了“肾-骨-髓-脑”的系统关联,是防治POP的重要理论依据。该文旨在以“肾-骨-髓-脑”系统理论为基础,从现代分子生物学角度探讨“肾-骨-髓-脑”系统与外泌体之间的关系,为POP的临床防治提供新思路。     

Pathophysiology‐based treatment of urolithiasis

Urolithiasis, a complex multifactorial disease, results from interactions between environmental and genetic factors. Epidemiological studies have shown the association of urolithiasis with a number of lifestyle‐related diseases, including cardiovascular diseases, hypertension, chronic kidney disease, diabetes and metabolic syndrome. Elucidation of the mechanisms underlying urinary stone formation will enable development of new preventive treatments. The present article reviews the epidemiology, pathophysiology and potential treatment of urolithiasis. Recent literature has shown that oxidative stress and reactive oxygen species could be one such mechanistic pathway. Calcium oxalate crystals adhering to renal tubular cells are incorporated into the cells through the involvement of osteopontin. Stimulation of crystal–cell adhesion impairs acceleration of the mitochondrial permeability transition pore in tubular cells, resulting in mitochondrial collapse, oxidative stress and activation of the apoptotic pathway in the initial steps of renal calcium crystallization. With regard to genetic factors, studies show that single nucleotide polymorphisms in genes encoding calcium‐sensing receptor, vitamin D receptor and osteopontin are correlated with urolithiasis. Genome‐wide association studies have shown that CLDN14 and NPT2 are associated with urolithiasis in Caucasian and Japanese populations, respectively. Thus, single nucleotide polymorphism analysis would aid in the prediction of urolithiasis risk and recurrence. New diagnostic methods and preventive approaches, along with complete removal of stones, will improve the management of urolithiasis.     

Oxidative stress, anti-oxidant therapies and chronic kidney disease

Chronic kidney disease (CKD) is a common and serious problem that adversely affects human health, limits longevity and increases costs to health-care systems worldwide. Its increasing incidence cannot be fully explained by traditional risk factors. Oxidative stress is prevalent in CKD patients and is considered to be an important pathogenic mechanism. Oxidative stress develops from an imbalance between free radical production often increased through dysfunctional mitochondria formed with increasing age, type 2 diabetes mellitus, inflammation, and reduced anti-oxidant defences. Perturbations in cellular oxidant handling influence downstream cellular signalling and, in the kidney, promote renal cell apoptosis and senescence, decreased regenerative ability of cells, and fibrosis. These factors have a stochastic deleterious effect on kidney function. The majority of studies investigating anti-oxidant treatments in CKD patients show a reduction in oxidative stress and many show improved renal function. Despite heterogeneity in the oxidative stress levels in the CKD population, there has been little effort to measure patient oxidative stress levels before the use of any anti-oxidants therapies to optimize outcome. This review describes the development of oxidative stress, how it can be measured, the involvement of mitochondrial dysfunction and the molecular pathways that are altered, the role of oxidative stress in CKD pathogenesis and an update on the amelioration of CKD using anti-oxidant therapies.     

白藜芦醇在绝经后骨质疏松症中抗氧化机制的研究进展

绝经后骨质疏松症(PO)是一种全身性骨骼疾病,由雌激素缺乏和衰老引起,其特征是低骨密度(BMD)和骨组织的微结构恶化,并导致骨质疏松性骨折的风险增高。在PO的发生发展过程中,潜在的致病机制是复杂和多方面的,雌激素缺乏是骨吸收增加和骨质流失加速的重要原因。同时,由于雌激素的缺乏和机体的衰老导致氧化应激(OS)水平的升高,是PO发生的主要机制之一。针对老年绝经后骨质疏松患者的治疗目前较为有效的是采用雌激素替代疗法,但也增加了患乳腺癌、子宫癌和心血管事件发生的风险。因此,寻找有效的抗氧化应激药物以治疗PO受到了越来越多研究者的关注。其中以白藜芦醇(RES)为代表的天然植物化合物,是一种具有抗氧化和抗衰老作用的天然抗氧化剂。它能通过多种通路抑制体内氧化应激,促进成骨分化和减少骨量丢失,并调节骨转换的能力,从而有效治疗PO。本文就白藜芦醇在PO中的抗氧化机制及其研究进展做一综述。     

Postconditioning is an effective strategy to reduce renal ischaemia/reperfusion injury

Background. Several recent studies have shown that a brief ischaemiaapplied during the onset of reperfusion (postconditioning) iscardioprotective in different animal models. The potential applicationof postconditioning to organs different from the heart, i.e.kidney, is not available and is investigated in the presentstudy. We also tested the hypothesis that mitochondria playa central role in renal protection during reperfusion. Methods. Wistar rats were subjected to left nephrectomy and90-min right kidney occlusion. In controls, the blood flow wasrestored without intervention. In postconditioned rats, completereperfusion was preceded by 3 min, 6 min and 12 min of reperfusionin a consecutive sequence, each separated by 5 min of reocclusion.Animals were studied for 48 h. Mitochondrial respiratory chainfunction, rate of hydroperoxide production and carbonyl proteinswere measured at the end of postconditioning and 24 h and 48h after reperfusion. Results. BUN and creatinine significantly decreased in the postconditioninggroup as compared to control rats. Mitochondrial respiratoryfunction was significantly impaired in control rats, mainlyat the level of Complex II. Postconditioning significantly reducedthis mitochondria impairment. The rate of mitochondrial peroxideproduction was higher in the control group than in the protectedgroup at the end of postconditioning reperfusion. Moreover,mitochondrial protein oxidation was significantly higher incontrol rats than in the postconditioning group at the end ofreperfusion. Conclusions. In the present study, postconditioning reducedrenal functional injury and reduces mitochondria respiratorychain impairment, mitochondria peroxide production and proteindamage.     

ApoE与骨质疏松症相关性的研究进展

骨质疏松症（osteoporosis,OP）作为一种比较常见的骨代谢疾病,与高血脂症间存在着一定的关联,临床上同时患有这两种疾病的现象比较常见。当骨吸收增加而骨形成减少时,骨代谢失衡,就会发生OP。载脂蛋白E（ApoE）是血浆中主要的载脂蛋白之一,ApoE功能异常是引起高血脂症的关键。近年来依托ApoE基因敲除鼠模型,在ApoE调节血脂机制及高脂血症与OP的发生过程相关性的研究有了诸多新发现,拓宽了固有认知。本文主要汇总了ApoE近5年与骨相关的研究,分析其影响OP的发生过程的机制,从宏观的血脂和细胞层面到微观的氧化应激和炎症层面,总结了ApoE对骨代谢的重要调节作用,以期为后续研究、新药物筛选及临床应用提供参考依据。     

Urinary F2-isoprostanes formation in kidney transplantation

BACKGROUND: Oxygen free-radical mediated lipid peroxidation has been implicated in many diseases such as chronic renal failure, hemodialysis and chronic kidney transplant rejection. However, insight into the role of free radical generation in kidney transplantation has been constrained by the limitations of current indexes of oxidant stress in vivo. Isoprostaglandin F2alpha type-III (iPF2alpha-III, formerly known as 8-iso-prostaglandin F2alpha) is emerging as a reliable marker of oxidant stress in vivo. The purpose of our study was to investigate iPF2alpha-III formation as an index of lipid peroxidation in the 5 d following kidney transplantation. METHODS: Urinary iPF2alpha-III measurements were performed by enzyme immunoassay from day I to 5 in 11 patients undergoing kidney transplantation. Results were compared with 11 healthy volunteers matched in sex, age and cigarette smoking. RESULTS: Urinary excretion of iPF2alpha-III at day 1 did not significantly differ between control and transplant group (111 +/- 17 vs. 92 +/- 10 pM/ mM creatinine, respectively, NS). Urinary iPF2alpha-III levels did not differ between day 1 to 5, and were not correlated to cold ischaemia time. CONCLUSION: Our study shows no evidence of enhanced lipid peroxidation in the first 5 d following kidney transplantation.     

绝经后骨质疏松症发病机制研究进展

绝经后骨质疏松症(postmenopausal osteoporosis,PMOP)是指女性绝经后卵巢内分泌功能失调衰退,导致雌激素水平下降,从而导致破骨细胞的骨吸收大于成骨细胞的骨形成的一种代谢性疾病。骨质疏松症没有特异性的临床表现,直到轻微的创伤诱发骨折时才会引起重视,所以预防骨质疏松症是临床工作的重中之重。目前有很多研究报道了骨质疏松症发生发展的机制,在机体内雌激素的缺乏引起炎症因子与MicroRNA激活,从而引起RANKL-RANK-OPG轴的紊乱,引起骨质流失。同时雌激素充当抗氧化剂来保护骨,抵抗氧化应激。本文就绝经后骨质疏松症的发生机制做一综述。     

The reduced tolerance of rat fatty liver to ischemia reperfusion is associated with mitochondrial oxidative injury

BACKGROUND: Oxidative stress contributes to the pathogenesis of hepatic ischemia-reperfusion injury. This study aimed to determine whether fatty degeneration affects the oxidative damage during warm ischemia reperfusion and whether mitochondria, the major intracellular site of energy synthesis, represent a preferential target of this injury. MATERIALS AND METHODS: Fed rats with control or fatty liver induced by choline deficiency underwent 60' lobar ischemia and reperfusion. Oxidative damage was assessed by measuring in whole liver tissue and in isolated mitochondria the thiobarbituric acid-reactive substances (TBARs), protein carbonyls (PC), and total and oxidized glutathione (GSH and GSSG) concentrations. The mitochondrial F0-F1-ATPase content and the oxidative phosphorylation activity were also determined. Rat survival and ALT release were assessed as parameters of liver injury. RESULTS: In the whole liver tissue, with the exception of TBARs, no differences were observed for GSH, GSSG, and PC between the two groups throughout all of the experiment. In contrast, in isolated mitochondria, fatty infiltration was associated with a mild oxidative imbalance already under basal conditions. The preischemic differences in the mitochondrial TBARs, PC, and GSSG levels were significantly amplified by reperfusion in the presence of steatosis. The enhanced oxidative damage was associated to a reduced F0-F1-ATPase content and oxidative phosphorylation activity in fatty liver mitochondria. Finally, serum ALT levels were significantly greater and survival significantly lower in rats with steatotic liver. CONCLUSIONS: Fatty infiltration exacerbates mitochondrial oxidative injury during warm ischemia reperfusion. The increased oxidative stress can alter mitochondrial functions, including key processes for ATP synthesis, thus, contributing to the reduced tolerance to reperfusion injury.     

Caveolins in the repair phase of acute renal failure after oxidative stress

Ischaemia-reperfusion and toxic injury are leading causes of acute renal failure (ARF). Both of these injury initiators use secondary mediators of damage in oxygen-derived free radicals. Several recent publications about ischaemia-reperfusion and toxin-induced ARF have indicated that plasma membrane structures called caveolae, and their proteins, the caveolins, are potential participants in protecting or repairing renal tissues. Caveolae and caveolins have previously been ascribed many functions, a number of which may mediate cell death or survival of injured renal cells. This review proposes possible pathophysiological mechanisms by which altered caveolin-1 expression and localization may affect renal cell survival following oxidative stress.     

Carnosine treatment during human semen processing by discontinuous density gradient

The aim of this article was to evaluate the effects of different concentrations of carnosine added during human semen processing. Semen samples from 34 patients were submitted to processing by discontinuous density gradient centrifugation without (control) or with different concentrations of carnosine supplementation as follows: (a) 20 mM of carnosine supplementation on the layers of Percoll; and (b) 50 mM carnosine supplementation. Sperm samples were then washed with human tubal fluid medium and evaluated according to sperm kinetics and functional assessment. For statistical analysis, data were evaluated by a general linear model or a Friedman test, whenever appropriate. The 50 mM carnosine supplementation led to improved sperm mitochondrial activity when compared to untreated samples. Motility variables, such as percentage of motile and progressively motile spermatozoa, average path velocity, straight line velocity, curvilinear velocity and linearity, showed an improvement after semen processing irrespective of carnosine supplementation. Both concentrations of carnosine increased the beat-cross frequency (BCF) when compared to samples before processing. We conclude that carnosine supplementation in semen samples benefits sperm mitochondrial activity and BCF.