自体吞噬与类固醇激素分泌调节的关系

本实验用腹腔内给予下丘脑激素或类固醇激素的方法，造成大鼠睾丸间质细胞和肾上腺皮质束状带细胞处于分泌兴奋或抑制状态，对这两种分泌类固醇激素细胞中的溶酶体和自噬小体进行了超微结构、细胞化学和形态计量研究。结果表明，在类固醇激素分泌增多时，细胞中自体吞噬活动减弱；激素分泌减少时，自体吞噬活动加强。这一结果证明了细胞内的自体吞噬活动与类固醇激素的分泌调节有密切关系，自体吞噬是溶酶体参与激素分泌调节过程的一种重要方式。     

中药单体调控自噬干预胚胎植入的研究进展

胚胎植入是生殖过程中最关键的步骤之一,植入失败的胚胎无法继续发育,是导致不孕的重要原因之一。胚胎植入的成功依赖于子宫内膜的高容受性和具有植入能力的胚胎。自噬是细胞质、细胞器和内含物被双膜囊泡吸收并运输到溶酶体进行降解和再循环的过程,是一种维持内环境稳态的方式。大量证据表明,自噬在胚胎植入的各个环节有着重要的作用。基于此探讨了自噬与子宫内膜容受性和胚胎植入能力的关系,并根据最新的研究进展,梳理了大黄素、梓醇、芍药苷、白藜芦醇、叶酸、玉米赤霉烯酮、姜黄素、汉黄芩素、槲皮素、白杨素、小檗碱、芹菜素、菲西汀、山柰酚在内的14种中药单体调控自噬干预胚胎植入的不同环节的5个机制,包括促进子宫内膜基质细胞蜕膜化、促进细胞凋亡、调节激素水平、协调炎症、促进排卵,希望对今后中药单体提高胚胎植入的成功率提供参考及思路。     

“肾虚络瘀，肾络癥瘕”病机观与肾间质纤维化中自噬不足的相关性

肾间质纤维化（RIF）是由多种病因导致慢性肾脏疾病的主要病理特征,结合中医“络病学说”“癥瘕理论”与现代医学肾脏病理特征,“肾虚络瘀,肾络癥瘕”是RIF的主要病机,正气亏虚,累及肾气,肾虚气化不利,痰热瘀毒等实邪阻滞肾络,相互胶结,形成癥瘕,积聚于肾络导致了RIF。自噬是细胞体内将受损或衰老的细胞器、变性的蛋白质等代谢产物进行降解与重吸收的过程,自噬参与了RIF发生的诸多环节,与RIF发生发展关系密切,将自噬与中医学结合研究发现,机体内生痰热瘀毒等实邪的代谢与自噬降解和重吸收功能相关,自噬在一定程度上是消除痰热瘀毒等实邪的方式,正气不足,累及肾气,肾虚气化不利,自噬功能不足,会引起痰热瘀毒等实邪累积,瘀阻肾络则易形成肾络癥瘕造成RIF,肾虚络瘀是自噬不足的基础,肾络癥瘕是自噬不足的微观病理表现,自噬不足与肾络癥瘕形成具有相同的病机演变,该文就RIF“肾虚络瘀,肾络癥瘕”病机,结合RIF-自噬病理机制,深入探讨“肾虚络瘀-自噬不足-肾络癥瘕”在RIF的相关性,全面诠释“肾虚络瘀,肾络癥瘕”病机的科学内涵,以期从中医理论阐述自噬在RIF中的作用研究奠定基础,为RIF的治疗及其机制研究提供新...     

基于“正虚邪滞”理论探讨肺癌自噬机制及中药干预进展

自噬是一种细胞自我保护和自我更新机制,与肺癌的发生发展密切相关,有利自噬可以减缓肺癌进展,而不利自噬能够促进肺癌进展,故调节自噬水平在肺癌治疗中具有重要意义。“正虚邪滞”是王永炎院士“虚气留滞”理论的延伸,是指因肺脾肾气亏虚,引发津液代谢异常,导致痰瘀阻滞的病理过程,以肺脾肾气亏虚为本、痰瘀阻滞为标,是肺癌的关键病机。肺癌的自噬机制与“正虚邪滞”有着相互贯通之处,肺脾肾气亏虚是肺癌有利自噬减弱的关键因素,进而抑制了肿瘤细胞凋亡,并导致有害物质蓄积;痰瘀阻滞是肺癌不利自噬增强的直接因素,进而促进了正常细胞自噬性死亡,削弱了免疫细胞对肿瘤细胞的免疫抑制作用,导致肿瘤细胞增殖和迁移。正虚与邪滞相合,致使自噬状态向着不利的方向不断发展,最终导致肺癌持续进展。因此,中医干预肺癌自噬机制需以扶正抗邪为原则,补益正气以治其本,祛痰化瘀以治其标,增强有利自噬的同时抑制不利自噬,将自噬水平调节到最佳状态,从而抑制肿瘤细胞增殖和迁移,促进肺癌的缓解。通过分析现有文献可知,目前通过自噬途径治疗肺癌的中医药以中药单体为主,中医药干预肺癌自噬的作用主要集中在促进自噬激活层面,这可能是因为藉由肺脾肾气亏虚引发的有利自噬减弱是肺癌发展的根本原因。     

Beneficial Effects of a Mixture of Algae and Extra Virgin Olive Oils on the Age-Induced Alterations of Rodent Skeletal Muscle: Role of HDAC-4

Aging is associated with a progressive decline in skeletal muscle mass, strength and function (sarcopenia). We have investigated whether a mixture of algae oil (25%) and extra virgin olive oil (75%) could exert beneficial effects on sarcopenia. Young (3 months) and old (24 months) male Wistar rats were treated with vehicle or with the oil mixture (OM) (2.5 mL/kg) for 21 days. Aging decreased gastrocnemius weight, total protein, and myosin heavy chain mRNA. Treatment with the OM prevented these effects. Concomitantly, OM administration decreased the inflammatory state in muscle; it prevented the increase of pro-inflammatory interleukin-6 (IL-6) and the decrease in anti-inflammatory interleukin-10 (IL-10) in aged rats. The OM was not able to prevent aging-induced alterations in either the insulin-like growth factor I/protein kinase B (IGF-I/Akt) pathway or in the increased expression of atrogenes in the gastrocnemius. However, the OM prevented decreased autophagy activity (ratio protein 1A/1B-light chain 3 (LC3b) II/I) induced by aging and increased expression of factors related with muscle senescence such as histone deacetylase 4 (HDAC-4), myogenin, and IGF-I binding protein 5 (IGFBP-5). These data suggest that the beneficial effects of the OM on muscle can be secondary to its anti-inflammatory effect and to the normalization of HDAC-4 and myogenin levels, making this treatment an alternative therapeutic tool for sarcopenia.     

Effects of Dietary Oat Beta-Glucans on Colon Apoptosis and Autophagy through TLRs and Dectin-1 Signaling Pathways—Crohn’s Disease Model Study

Background: Crohn’s disease (CD) is characterized by chronic inflammation of the gastrointestinal tract with alternating periods of exacerbation and remission. The aim of this study was to determine the time-dependent effects of dietary oat beta-glucans on colon apoptosis and autophagy in the CD rat model. Methods: A total of 150 Sprague–Dawley rats were divided into two main groups: healthy control (H) and a TNBS (2,4,6-trinitrobenzosulfonic acid)-induced colitis (C) group, both including subgroups fed with feed without beta-glucans (βG−) or feed supplemented with low- (βGl) or high-molar-mass oat beta-glucans (βGh) for 3, 7, or 21 days. The expression of autophagy (LC3B) and apoptosis (Caspase-3) markers, as well as Toll-like (TLRs) and Dectin-1 receptors, in the colon epithelial cells, was determined using immunohistochemistry and Western blot. Results: The results showed that in rats with colitis, after 3 days of induction of inflammation, the expression of Caspase-3 and LC3B in intestinal epithelial cells did not change, while that of TLR 4 and Dectin-1 decreased. Beta-glucan supplementation caused an increase in the expression of TLR 5 and Dectin-1 with no changes in the expression of Caspase-3 and LC3B. After 7 days, a high expression of Caspase-3 was observed in the colitis-induced animals without any changes in the expression of LC3B and TLRs, and simultaneously, a decrease in Dectin-1 expression was observed. The consumption of feed with βGl or βGh resulted in a decrease in Caspase-3 expression and an increase in TLR 5 expression in the CβGl group, with no change in the expression of LC3B and TLR 4. After 21 days, the expression of Caspase-3 and TLRs was not changed by colitis, while that of LC3B and Dectin-1 was decreased. Feed supplementation with βGh resulted in an increase in the expression of both Caspase-3 and LC3B, while the consumption of feed with βGh and βGl increased Dectin-1 expression. However, regardless of the type of nutritional intervention, the expression of TLRs did not change after 21 days. Conclusions: Dietary intake of βGl and βGh significantly reduced colitis by time-dependent modification of autophagy and apoptosis, with βGI exhibiting a stronger effect on apoptosis and βGh on autophagy. The mechanism of this action may be based on the activation of TLRs and Dectin-1 receptor and depends on the period of exacerbation or remission of CD.     

氯化锂通过TGFBI促进角膜基质成纤维细胞增殖和自噬

目的：研究TGFBI和微管相关蛋白轻链3(LC3)在角膜营养不良患者中的表达,及氯化锂(LiCl)通过TGFBI对角膜基质成纤维细胞增殖能力的影响。

方法：用免疫组化和Western-blot方法检测角膜营养不良及正常角膜组织中TGFBI和LC3的表达。实验构建了TGFBI过表达载体并转染角膜基质成纤维细胞,分别以5、10、20、40mmol/L LiCl作用于突变型TGFBI转染的角膜基质成纤维细胞,检测不同时间(0、1、6、12h)后,TGFBI与LC3蛋白表达变化,并用CCK-8法检测细胞增殖活性。

结果：TGFBI和LC3在角膜营养不良患者角膜组织中显著高表达。TGFBI过表达抑制角膜基质成纤维细胞增殖活性(P<0.05)。LiCl抑制突变型TGFBI转染的角膜基质成纤维细胞中TGFBI和LC3蛋白表达,并增强其细胞增殖活性(P<0.05)。

结论：LiCl可以促进角膜基质成纤维细胞增殖活性和自噬,其作用机制与下调TGFBI和LC3的表达有关。     

mTOR通路对神经母细胞瘤SH-SY5 Y细胞自噬作用的影响

目的：研究雷帕霉素靶蛋白（ mammalian target of rapamycin,mTOR）通路对神经母细胞瘤SH-SY5Y细胞自体吞噬作用的影响。方法：应用不同浓度的雷帕霉素作用于神经母细胞瘤细胞,采用实时荧光定量PCR检测雷帕霉素作用前后神经母细胞瘤细胞株的mTOR、LC3及Beclin 1 mRNA表达情况。结果：雷帕霉素能抑制mTOR表达且呈现量效依赖关系,不同浓度的雷帕霉素分别对SH-SY5Y细胞作用72h后,mTOR mR-NA的表达随浓度增加逐渐下降。而LC3及Beclin 1 mRNA的表达随浓度增加逐渐上升,神经母细胞瘤中mTOR与LC3及Beclin 1的表达呈负相关。结论：雷帕霉素通过抑制mTOR信号通路促进神经母细胞瘤的自体吞噬作用,动态观察mTOR可作为神经母细胞瘤治疗效果监测的指标之一。     

自噬在阿尔茨海默病中的作用及中药的干预作用

对近年来阿尔茨海默病(Alzheimer's disease,AD)与自噬相关性的研究及中药对自噬的调控作用进行介绍,对自噬的过程和相关分子机制、信号途径,自噬参与AD病理学表现的形成、神经元的缺失及与其他AD病理学表现,中药对自噬的影响,自噬在AD中的治疗意义进行总结.研究表明,自噬功能缺陷与多种神经退行性疾病密切相关,包括AD.集中介绍自噬在AD发生发展过程中的作用以及调控自噬在阻止或延缓AD发生,改善临床症状等方面的潜在治疗价值.在调节自噬,治疗AD等神经退行性疾病方面,关于中药的研究几乎处于空白状态.研究中药对神经细胞自噬的影响,通过调节自噬探讨中药治疗AD的潜在可能,并以此为基础,进行复方药物的选择,具有重要意义和极大的研究潜力.     

过表达miR-520d 通过抑制自噬蛋白Beclin1 逆转三阴性乳腺癌细胞化疗耐药性

[摘要] 目的：探讨miR-520d 通过调控自噬逆转三阴性乳腺癌（TNBC）细胞化疗耐药的作用及分子机制。方法：以人TNBC细胞系MDA-MB-231 和MDA-MB-468 为亲本株细胞构建多西他赛（Doc）耐药细胞株MDA-MB-231/Doc 和MDA-MB-468/Doc,实验分为空白组（亲本细胞）、对照组（耐药细胞组）和过表达miR-520d 组。用qPCR检测空白组和耐药细胞组细胞中miR-520d 的表达水平,MTT实验检测过表达miR-520d 的耐药细胞对Doc 的敏感性,MDC染色后荧光显微镜观察细胞中自噬小体的发生情况、共聚焦显微镜观察过表达miR-520d 的耐药细胞中自噬相关蛋白LC3 阳性的细胞数。用荧光素酶报告基因实验验证miR-520d 与Beclin1 的靶向关系。用WB实验检测过表达miR-520 对细胞中自噬相关蛋白Beclin1 和LC3Ⅰ、LC3Ⅱ表达的影响。结果：TNBC耐药细胞中miR-520d 的表达水平明显低于空白组细胞（P<0.01）。过表达miR-520d 的耐药细胞对Doc的敏感性显著提高（P<0.01）、细胞的自噬活性明显降低（P<0.01）。荧光素酶报告基因实验证明Beclin1 是miR-520d 可能的靶分子。Doc 与miR-520d mimics 联合使用可降低TNBC耐药细胞中LC3-Ⅱ/Ⅰ比值和自噬蛋白Beclin1 的表达水平（均P<0.05）。结论：通过调控miR-520d 水平可能改变自噬蛋白Beclin1 表达,从而逆转TNBC细胞Doc化疗耐药性。