Antimicrobial Effectiveness on Selected Bacterial Species and Alkaloid and Saponin Content of <i>Rosa nutkana</i>C. Presl (Nootka Rose) and <i>Urtica dioica</i>L. (Stinging Nettle) Extracts

Nootka rose (Rosa nutkana C. Presl) and stinging nettle (Urtica dioica L.) have been traditionally used in the treatment of skin infection by Indigenous peoples of Vancouver Island, British Columbia, Canada. The main objective of this study was to examine the antibacterial efficacy of extracts of Nootka rose and stinging nettle against the common pathogenic skin bacteria Staphylococcus aureus, Micrococcus luteus, and Pseudomonas aeruginosa using Indigenous science and standard methods of analysis. The Indigenous science method of plant extraction by steeping as advised by the Traditional Knowledge keeper was performed to examine minimum inhibitory concentration (MIC) values and minimum bactericidal concentrations (MBC) by serial dilution and bacterial population counts. Soxhlet extractions and Kirby Bauer disc sensitivity testing showed that Nootka rose extracts possessed antibacterial effectiveness against all three bacterial species while stinging nettle extracts were effective against M. luteus. Results for MIC and MBC indicated antibacterial activity against M. luteus and S. aureus for the Nootka rose when using full-strength solutions;all three bacterial species exhibited growth when undiluted stinging nettle treatments were used. When considering bacterial population counts for S. aureus, results indicated that only the Nootka rose treatment offered effective inhibition. Chemical analysis showed that alkaloid percentage was greater in the stinging nettle (0.17%) than Nootka rose (0.07%), while saponin percentage was greater in the Nootka rose (0.87%) than stinging nettle (0.17%). Overall, Nootka rose showed a greater level of antibacterial effectiveness than stinging nettle by Indigenous and Western scientific methods of plant extract preparation.

Effects of stinging nettle (Urtica dioica) powder on laying performance,egg quality,and serum biochemical parameters of Japanese quails

The aim of this experiment was to evaluate the effects of dietary supplementation of stinging nettle powder(SNP) on laying performance,egg quality,and some selected serum biochemical parameters of quails.One hundred and forty-four 10-wk-old Japanese quails(initial body weight=199±18 g) were divided into 3 dietary treatment groups(basic diet without SNP [SNPO],SNPO with 3% SNP [SNP3],SNPO with 6% SNP [SNP6]) with 4 replicates of 12 quails for a rearing period of 12 wk.At 22 wk of age,the final body weights of the SNP3 and SNP6 groups were significantly(P=0.001) reduced compared to that of the SNPO group.Daily feed intake was not statistically different among the groups.The mean number of eggs laid ranged from 65 to 69 with laying rates from 76.8% to 82.1%.The percentage of cracked eggs was not significantly different among the groups and ranged from 1.6% to 1.9%.The egg weight was similar and the feed conversion ratio was closer among the groups.The egg yolk cholesterol,serum cholesterol and serum triglyceride levels in the SNP6 group were significantly reduced(P <0.001) compared to those of the SNPO group.Serum Ca,P and Mg were not significantly influenced by the supplementation.In conclusion,the results demonstrated that the supplementation of SNP to the quail diet at the level of 6%reduced quail egg yolk cholesterol,serum total cholesterol and serum triglyceride levels and did not negatively influence quail performance.

Pharmacological targeting cGAS/STING/NF-κB axis by tryptanthrin induces microglia polarization toward M2 phenotype and promotes functional recovery in a mouse model of spinal cord injury

The M1/M2 phenotypic shift of microglia after spinal cord injury plays an important role in the regulation of neuroinflammation during the secondary injury phase of spinal cord injury.Regulation of shifting microglia polarization from M1(neurotoxic and proinflammatory type)to M2(neuroprotective and anti-inflammatory type)after spinal cord injury appears to be crucial.Tryptanthrin possesses an anti-inflammatory biological function.However,its roles and the underlying molecular mechanisms in spinal cord injury remain unknown.In this study,we found that tryptanthrin inhibited microglia-derived inflammation by promoting polarization to the M2 phenotype in vitro.Tryptanthrin promoted M2 polarization through inactivating the cGAS/STING/NF-κB pathway.Additionally,we found that targeting the cGAS/STING/NF-κB pathway with tryptanthrin shifted microglia from the M1 to M2 phenotype after spinal cord injury,inhibited neuronal loss,and promoted tissue repair and functional recovery in a mouse model of spinal cord injury.Finally,using a conditional co-culture system,we found that microglia treated with tryptanthrin suppressed endoplasmic reticulum stress-related neuronal apoptosis.Taken together,these results suggest that by targeting the cGAS/STING/NF-κB axis,tryptanthrin attenuates microglia-derived neuroinflammation and promotes functional recovery after spinal cord injury through shifting microglia polarization to the M2 phenotype.

The cGAS-STING-interferon regulatory factor 7 pathway regulates neuroinflammation in Parkinson's disease

Interferon regulatory factor 7 plays a crucial role in the innate immune response.However,whether interferon regulatory factor 7-mediated signaling contributes to Parkinson's disease remains unknown.Here we report that interferon regulatory factor 7 is markedly up-regulated in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced mouse model of Parkinson's disease and co-localizes with microglial cells.Both the selective cyclic guanosine monophosphate adenosine monophosphate synthase inhibitor RU.521 and the stimulator of interferon genes inhibitor H151 effectively suppressed interferon regulatory factor 7 activation in BV2 microglia exposed to 1-methyl-4-phenylpyridinium and inhibited transformation of mouse BV2 microglia into the neurotoxic M1 phenotype.In addition,si RNA-mediated knockdown of interferon regulatory factor 7 expression in BV2 microglia reduced the expression of inducible nitric oxide synthase,tumor necrosis factorα,CD16,CD32,and CD86 and increased the expression of the anti-inflammatory markers ARG1 and YM1.Taken together,our findings indicate that the cyclic guanosine monophosphate adenosine monophosphate synthase-stimulator of interferon genes-interferon regulatory factor 7 pathway plays a crucial role in the pathogenesis of Parkinson's disease.

Inhibition of the cGAS–STING pathway:contributing to the treatment of cerebral ischemia-reperfusion injury

The cGAS–STING pathway plays an important role in ischemia-reperfusion injury in the heart,liver,brain,and kidney,but its role and mechanisms in cerebral ischemia-reperfusion injury have not been systematically reviewed.Here,we outline the components of the cGAS–STING pathway and then analyze its role in autophagy,ferroptosis,cellular pyroptosis,disequilibrium of calcium homeostasis,inflammatory responses,disruption of the blood–brain barrier,microglia transformation,and complement system activation following cerebral ischemia-reperfusion injury.We further analyze the value of cGAS–STING pathway inhibitors in the treatment of cerebral ischemia-reperfusion injury and conclude that the pathway can regulate cerebral ischemia-reperfusion injury through multiple mechanisms.Inhibition of the cGAS–STING pathway may be helpful in the treatment of cerebral ischemia-reperfusion injury.

Effect of an amalgamated antibiotic and its connection to cytogenotoxicity and histo-architectural malformations in stinging catfish

Intensive fish farming has an excessive prevalence of infection and is typically controlled by the administration of antibiotics.Although amalgamated antibiotics are a relatively novel therapeutic idea and more effective than traditional antibiotic monotherapy,they can also have a toxic effect on the fish body when it is administered abruptly.This study investigated the cyto-genotoxic effects on erythrocytes and histo-architectural malformations in the liver and kidneys of stinging catfish(Heteropneustes fossilis)in relation to Enrocip plus use as an amalgamated antibiotic agent.The experimental fish with an initial average weight of 17.38±1.94 g were divided into four treatment groups with antibiotic doses:according to the recommendation of the manufacturer 0.167 mg/ml Enrocip plus was used as a standard dose(x)and it was treated as T3,while 0%of the standard dose(0×mg/ml),1/2×(0.083 mg/ml),and 2×(0.333 mg/ml)were treated as T1,T2 and T4,respectively for a period of 30 days.The observed erythrocyte cellular deformities(ECD)were twin,tear-drop,serrated,tail budded and de-membranated cells,whereas the erythrocyte nuclear deformities(END)were a nuclear bridge,bi-nucleus,nuclear termination,karyopyknosis,and micronucleus at different concentrations of Enrocip plus.Both ECD and END percentages experienced the interaction of antibiotic dose and exposure time,and were significantly different(P<0.01).Significant changes in hepatocytes,mild to severe necrosis,vacuole formation,and hepatopancreas damage were also observed in the liver of the treated fish whereas highly degraded renal tubules and hematopoietic tissue,glomerular occlusion,and vacuolation were evident in the kidneys.The current investigation fully emphasizes the adverse effects of amalgamated antibiotics on the cytogenotoxicity and the histomorphology of the kidneys and liver of fish.Thus,the use of an amalgamated antibiotic in aquaculture must be carefully evaluated.

瑞香素调节STING/TBK1/IRF3信号通路对脓毒症大鼠免疫炎症反应的影响

目的探究瑞香素调节STING/TBK1/IRF3信号通路对脓毒症大鼠免疫炎症反应的影响。方法选用80只雄性Sprague Dawley(SD)大鼠,将大鼠随机分为4组,假手术组20只;其余60只按照盲肠结扎穿刺法(CLP)构建脓毒症小鼠模型,腹腔注射不同剂量瑞香素,分为无瑞香素模型组(0 mg/kg)、低剂量组(2.5 mg/kg)、高剂量组(5 mg/kg),每组20只;模型组及假手术组腹腔注射等量0.1%DMSO。观察CLP建模后的各组大鼠存活率。收集肺、肾、肝和脾组织进行病理组织学分析。ELISA检测大鼠血清炎症因子(TNF-α、IL-6、IL-1β),流式细胞术检测外周血中的T淋巴细胞亚群,Western blotting检测STING/TBK1/IRF3信号通路蛋白表达变化。结果与假手术组相比,模型组大鼠生存率降低(P<0.05),各个脏器组织损伤明显,血清炎症因子(TNF-α、IL-6、IL-1β)水平上升(P<0.05),外周血CD4^(+)比例、CD4^(+)/CD8^(+)比值均降低(P<0.05),CD8^(+)比例、Th1/Th2比值均升高(P<0.05),STING、p-TBK1/TBK1、p-IRF3/IRF3蛋白表达均下调(P<0.05)。与模型组相比,瑞香素高剂量组提高了大鼠生存率(P<0.05),各脏器组织损伤明显减轻,血清炎症因子水平降低(P<0.05),外周血CD4^(+)比例、CD4^(+)/CD8^(+)比值升高(P<0.05),CD8^(+)比例、Th1/Th2比值降低(P<0.05),STING、p-TBK1/TBK1、p-IRF3/IRF3蛋白表达升高(P<0.05)。结论瑞香素可通过调节STING/TBK1/IRF3信号通路,改善脓毒症大鼠免疫炎症反应。

易黄汤对脾虚湿热型宫颈癌小鼠皮下移植瘤cGAS/STING/IRF-3信号通路及相关免疫因子的影响

目的探讨易黄汤对脾虚湿热型宫颈癌小鼠皮下移植瘤环磷酸鸟苷-腺苷酸合成酶(cGAS)/干扰素刺激基因(STING)/干扰素调节因子-3(IRF-3)信号通路及相关免疫因子的影响。方法选取6~8周龄SPF级雌性BALB/c小鼠50只,随机分为造模组40只和空白组10只。空白组正常喂养,造模组通过湿热环境及高脂高糖饮食建立脾虚湿热型小鼠模型,当小鼠出现嗜卧懒动、易激惹、形体消瘦、进食及饮水量减少、毛发疏松粗糙、粪便时干时溏、肛温稍有增加则认为造模成功。将2×106个/mL对数生长期HPV16阳性人宫颈癌CaSki细胞悬液0.2 mL接种于上述50只小鼠右腋下,5~7 d后可触及直径≥5 mm结节时即认为皮下移植瘤造模成功。将造模组按随机数字表法分为模型组和易黄汤低剂量、中剂量、高剂量组,每组10只。低、中、高剂量组均按体质量40.0 mL/kg分别予含生药浓度0.25、0.50、1.00 g/mL易黄汤药液灌胃,模型组和空白组予同等剂量生理盐水灌胃,均隔日1次,干预4周。干预后计算5组小鼠胸腺、脾脏指数和瘤质量、抑瘤率;ELISA法检测5组小鼠血清白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平;支链DNA信号扩增法检测5组小鼠皮下移植瘤组织HPV E6 mRNA转录水平;Western blot测定5组小鼠移植瘤组织中cGAS、STING、IRF-3蛋白表达量。结果与空白组比较,模型组胸腺和脾脏指数、IL-6、TNF-α水平显著下降(P<0.05),HPV E6 mRNA转录水平显著增加(P<0.05);易黄汤不同剂量组中以高剂量组的抑瘤率最高为(35.42±4.86)%(P<0.05);与模型组比较,低、中、高剂量组胸腺和脾脏指数、IL-6、TNF-α水平及cGAS、STING和IRF-3蛋白表达量显著增加(P<0.05),皮下移植瘤质量及HPV E6 mRNA转录水平显著降低(P<0.05),且降低程度与易黄汤剂量成正比。结论脾虚湿热能促进CaSki细胞生长、HPV表达增加,易黄汤可能通过激活cGAS/STING/IRF-3信号通路增强免疫功能,从而抑制HPV表达及宫颈癌小鼠移植瘤生长。

cGAS-STING信号通路调节剂在免疫治疗中的研究进展

环鸟嘌呤-腺嘌呤核苷酸合成酶(cGAS)-干扰素基因刺激蛋白(STING)信号通路感知细胞质中的异常双链DNA后,诱导Ⅰ型干扰素(IFN-Ⅰ)和促炎细胞因子表达,从而激活宿主的免疫应答,增强机体抗肿瘤免疫反应和抗病原体感染。但是,cGAS-STING信号通路的持续激活会驱动自身免疫性疾病、衰老相关炎症和神经退行性病变等疾病。本文阐述了cGAS-STING信号通路参与调控多种免疫相关性疾病发生发展的机制,重点回顾了STING激动剂、cGAS抑制剂以及STING抑制剂的研发进展,为cGAS-STING调节剂的研发提供更多理论参考。

干扰素基因刺激因子激动剂的研究进展

cGAS-STING通路作为固有免疫系统的重要组成部分,可以识别来自病原微生物和宿主细胞的胞质DNA,诱导Ⅰ型干扰素的分泌和炎症细胞因子的生成,激活机体免疫系统,抵抗病原微生物的入侵及增强机体抗肿瘤活性等。该文简要介绍了cGAS-STING通路的生物学功能,主要聚焦于STING激动剂的研究进展,包括CDN类和Non-CDN类STING激动剂的药理作用及其研究阶段,总结与展望STING激动剂研发过程中的问题与挑战,以期为后续相关药物的研发和疾病治疗提供思路。

NLRC3通过抑制STING信号通路, 减轻类风湿关节炎患者巨噬细胞焦亡诱导的免疫炎症反应

目的探索含胱天蛋白酶激活和募集结构域核苷酸结合寡聚结构域样受体3(NLRC3)在RA患者巨噬细胞焦亡诱导的免疫炎症反应中的作用及潜在的机制。方法按照入组标准选取50名类风湿关节炎(RA)患者和10名健康志愿者,提取外周血巨噬细胞,根据分组要求进行转染处理,分为正常健康对照(NC)组、RA巨噬细胞模型(RA-MC)组、RA巨噬细胞模型联合NLRC3过表达(RA-MC联合pcDNA3.1-NLRC3)组、RA巨噬细胞模型联合NLRC3敲低(RA-MC联合siRNA-NLRC3)组、RA巨噬细胞模型联合干扰素基因刺激因子(STING)过表达(RA-MC联合pcDNA3.1-STING)组、RA巨噬细胞模型联合STING敲低(RA-MC联合siRNA-STING)组,采用透射电镜观察各组巨噬细胞焦亡的情况,实时定量PCR检测各组巨噬细胞中NLRC3、STING、胱天蛋白酶1(caspase-1)、消皮素D(GSDMD)的mRNA表达,ELISA测定各组巨噬细胞上清液中白细胞介素1β(IL-1β)、IL-18的水平。结果与NC组相比,RA-MC组中巨噬细胞表现出细胞焦亡的特点;与RA-MC组相比,RA-MC联合pcDNA3.1-NLRC3组和RA-MC联合siRNA-STING组中细胞焦亡情况改善,RA-MC联合siRNA-NLRC3组和RA-MC联合pcDNA3.1-STING组中细胞焦亡情况加重;与NC组相比,RA-MC组中STING、caspase-1、GSDMD的mRNA相对表达水平升高,炎症因子IL-1β、IL-18的水平也升高,而NLRC3的mRNA相对表达水平降低;与RA-MC组相比,RA-MC联合pcDNA3.1-NLRC3组和RA-MC联合siRNA-STING组中caspase-1、GSDMD的mRNA相对表达水平降低,炎症因子IL-1β、IL-18的水平也降低,RA-MC联合siRNA-NLRC3组和RA-MC联合pcDNA3.1-STING组的情况则与之相反;同样与RA-MC组相比,RA-MC联合pcDNA3.1-NLRC3组中STING的mRNA相对表达水平会降低,RA-MC联合siRNA-NLRC3组中STING的mRNA相对表达水平升高。结论NLRC3可通过抑制STING信号通路,减少caspase-1、GSDMD焦亡蛋白的产生,进而拮抗巨噬细胞焦亡,降低炎症因子IL-1β、IL-18的水平,从而减轻RA患者的免疫炎症反应。

桂枝芍药知母汤抑制cGAS/STING/NF-κB通路对类风湿关节炎大鼠炎症活动的影响

目的基于环状GMP-AMP合成酶(cGAS)/干扰素基因刺激因子(STING)/核因子-κB(NF-κB)通路探讨桂枝芍药知母汤对类风湿关节炎大鼠炎症活动的影响及机制。方法取70只Wistar大鼠,随机选择10只作为正常组,其余60只大鼠制备胶原诱导性关节炎模型。将造模成功大鼠随机分为模型组、甲氨蝶呤组、雷公藤组及桂芍知母汤低、中、高剂量组,每组10只。甲氨蝶呤组给予0.104 mg/mL的甲氨蝶呤片混悬液灌胃,1次/周,其余时间每天给予蒸馏水灌胃1次;雷公藤组给予0.625 mg/mL的雷公藤多甙片混悬液灌胃,桂芍知母汤低、中、高剂量组分别给予0.958 g/mL、1.916 g/mL、2.874 g/mL的桂枝芍药知母汤灌胃,正常组、模型组给予蒸馏水灌胃,均1次/d。各组均干预4周。测量各组大鼠干预1,2,3,4周末足趾肿胀程度,并计算关节炎指数(AI);末次干预结束后,ELISA法检测血清白细胞介素-1β(IL-1β)、白细胞介素-17(IL-17)、肿瘤坏死因子-α(TNF-α)水平,Western blot法检测滑膜组织中cGAS、STING、NF-κB p65、IκBα蛋白表达情况。结果干预1,2,3,4周末,模型组大鼠的关节肿胀度均明显高于正常组(P均<0.05);随时间推移,模型组大鼠的关节肿胀度逐渐增加,各药物干预组大鼠的关节肿胀度有不同程度的降低,其中桂芍知母汤中剂量组干预4周末的关节肿胀度明显低于模型组(P<0.05)。干预1周末,各组间AI评分比较差异均无统计学意义(P均>0.05);干预2,3,4周末,桂芍知母汤高剂量组AI评分均明显低于模型组和桂芍知母汤低剂量组(P均<0.05),且桂芍知母汤高剂量组干预2周末的AI评分明显低于雷公藤组(P<0.05);干预3周末,甲氨蝶呤组、桂芍知母汤中剂量组AI评分均明显低于模型组(P均<0.05);干预4周末,甲氨蝶呤组、雷公藤组、桂芍知母汤中剂量组、桂芍知母汤低剂量组AI评分均明显低于模型组(P均<0.05)。与正常组比较,模型组血清IL-1β、IL-17、TNF-α水平及滑膜组织中cGAS、STING、NF-κB p65蛋白相对表达量均明显升高(P均<0.05),滑膜组织中IκBα蛋白相对表达量明显降低(P<0.05);与模型组比较,各药物干预组血清IL-1β、IL-17、TNF-α水平及滑膜组织中cGAS、STING、NF-κB p65蛋白相对表达量均明显降低(P均<0.05),滑膜组织中IκBα蛋白相对表达量均明显升高(P均<0.05)。桂芍知母汤高剂量组和甲氨蝶呤组血清IL-1β、IL-17和TNF-α水平及滑膜组织中cGAS、STING、NF-κB p65蛋白相对表达量均明显低于桂芍知母汤低剂量组(P均<0.05),滑膜组织中IκBα蛋白相对表达量均明显高于桂芍知母汤低剂量组(P均<0.05)。结论桂枝芍药知母汤可明显减轻类风湿关节炎大鼠关节肿胀和关节炎症,机制可能与抑制cGAS/STING/NF-κB通路,降低炎症因子水平有关。

车叶草苷减轻溃疡性结肠炎大鼠结肠黏膜损伤及炎症反应的机制

目的基于干扰素基因刺激因子(STING)/TANK结合激酶1(TBK1)/干扰素调节因子3(IRF3)信号通路,探究车叶草苷(ASP)对溃疡性结肠炎(UC)大鼠结肠组织病理损伤及炎症反应的影响机制。方法以肠腔内注射三硝基苯磺酸和乙醇构建UC大鼠模型,并将造模成功的大鼠分为模型组、ASP低剂量组(17.5 mg/kg)、ASP高剂量组(35 mg/kg)、ASP高剂量+STING激活剂组(35 mg/kg ASP+20 mg/kg ADU-S100),每组16只;另取16只健康大鼠,肠腔内注射生理盐水,作为对照组。各组大鼠灌胃或/和腹腔注射相应药液或生理盐水,每天1次,连续14 d。末次给药后24 h,使用疾病活动指数(DAI)和结肠黏膜损伤指数(CMDI)分别评估各组大鼠的UC严重程度和结肠黏膜损伤程度,观察其结肠组织病理改变并进行组织病理学评分,检测其结肠组织细胞凋亡情况、炎症因子[肿瘤坏死因子α(TNF-α)、β干扰素(IFN-β)、白细胞介素4(IL-4)、IL-10]水平和通路相关蛋白表达情况[STING、TBK1、IRF3、核因子κB p65(NF-κB p65)]。结果与对照组比较,模型组大鼠结肠组织黏膜及腺体结构严重受损,黏膜上皮磨损,隐窝缺失,可见明显的炎症细胞浸润;DAI、CMDI、结肠组织病理学评分,细胞凋亡率,TNF-α、IFN-β水平,以及STING蛋白的表达水平和TBK1、IRF3、NF-κB p65蛋白的磷酸化水平均显著升高;IL-4、IL-10水平均显著降低(P<0.05)。与模型组比较,ASP低、高剂量组大鼠结肠组织黏膜结构相对完整,腺体排列有序,充血和水肿减轻,炎症细胞浸润及溃疡明显减少;上述各定量指标均显著改善,且高剂量组的改善较低剂量组明显(P<0.05)。与ASP高剂量组比较,ASP高剂量+STING激活剂组大鼠的上述指标均被显著逆转(P<0.05)。结论ASP可能通过抑制STING/TBK1/IRF3信号通路来减轻UC大鼠的结肠组织病理损伤及炎症反应。

STING信号通路在骨质疏松中作用的研究进展

目的概述STING信号通路在骨质疏松中的作用机制与研究成果,从而为骨质疏松的治疗提供新的思路。方法于2023年11月在PubMed和中国知网数据库中以“STING、osteoporosis、osteoblast、osteoclast、IFN-β、NF-κB、type H vessels”为英文检索词进行检索,以“STING信号通路、成骨细胞、破骨细胞、IFN-β、NF-κB、H型血管”为中文检索词进行检索。结果通过阅读标题及摘要进行文献筛选,最终引用论文数41篇。结论STING通路为骨质疏松症中的治疗提供了新的方向。激活STING/IFN-β信号通路可抑制破骨细胞分化,抑制骨吸收。而STING/NF-κB信号可导致骨吸收增加、骨形成减少。此外,STING信号通路的激活可抑制具有成骨能力的H型血管的产生,从而抑制骨形成。因此,STING通路在骨质疏松疾病进展中具有双重调控作用,调控STING通路的不同分支有望发展更为有效的骨质疏松症治疗方法。

心康冲剂基于cGAS/STING信号通路对慢性心力衰竭大鼠炎症因子调控作用的研究

目的基于cGAS/STING信号通路探讨心康冲剂对慢性心力衰竭大鼠炎症因子的干预作用及分子机制。方法将SD大鼠随机分为正常组、造模组,通过腹腔注射盐酸阿霉素建立慢性心力衰竭模型,待模型成功后再分为模型组、缬沙坦组、心康冲剂组。模型组每日灌胃蒸馏水,缬沙坦组每日以缬沙坦溶液灌胃,心康冲剂组每日灌胃心康冲剂溶液治疗,连续4周。采用超声心动图检测各组大鼠心功能;苏木素-伊红染色(HE)检测各组大鼠心肌病理学改变;透射电镜观察各组大鼠心肌超微结构改变;采用酶联免疫吸附测定法(ELISA)检测各组大鼠血清中白细胞介素1β(IL-1β)、白细胞介素6(IL-6)含量;实时荧光定量法(qPCR)检测各组大鼠心肌组织中线粒体转录因子A(TFAM)、环化鸟苷酸-腺苷酸合成酶(cGAS)、干扰素刺激基因(STING)、IL-6 mRNA的表达水平。免疫组化法检测大鼠心肌组织cGAS、STING蛋白表达。结果与空白组比较,模型组大鼠心肌炎性细胞浸润明显,可见炎性水肿;心功能明显降低(P<0.05,P<0.01),血清炎症因子明显升高(P<0.01),心肌组织中TFAM mRNA表达明显降低(P<0.01),IL-6、cGAS、STING mRNA表达明显上调(P<0.01),心肌组织中cGAS、STING蛋白明显增加(P<0.01)。与模型组比较,心康冲剂组大鼠心功能明显改善(P<0.05,P<0.01);心肌细胞炎性浸润减轻;血清炎性因子表达明显降低(P<0.01),心肌组织中TFAM mRNA表达明显增加(P<0.05),IL-6、cGAS、STING mRNA表达明显下调(P<0.01);心肌组织中cGAS、STING蛋白表达明显下调(P<0.01)。结论心康冲剂可降低慢性心力衰竭大鼠炎性因子表达,改善心功能,其机制可能与抑制cGAS/STING信号通路有关。

GV1001抑制cGAS/STING通路缓解阿霉素诱导的小鼠心脏毒性

目的:探讨端粒酶衍生肽GV1001对阿霉素(DOX)诱导的小鼠心脏毒性的保护作用及其潜在机制。方法:使用C57BL/6小鼠建立DOX心脏毒性动物模型,分为对照组、DOX组(20 mg/kg)与不同剂量GV1001(6、30和60μmol/kg)组。检测各组小鼠心功能,取血清行心肌损伤标志物检测;取心脏组织进行HE染色及Masson染色;采用分光光度法测定心肌MDA、T-SOD和GSH-PX水平;提取心脏组织蛋白,Western blot法检测环状GMP-AMP合成酶(cGAS)、磷酸化干扰素基因刺激因子(STING)和干扰素调节因子3(IRF3)蛋白的表达水平。并使用STING激动剂后检测心肌组织损伤和氧化应激的改变。结果:与对照组比,DOX组小鼠LVIDd、LVEF和LVFS降低,LVIDs值增加,心肌纤维排列紊乱,细胞核固缩、空泡化,胶原沉积增加(均P<0.05);同时血清心肌酶学指标cTnI、CK-MB和LDH升高,心肌组织MDA增加,而T-SOD和GSH-PX下降(均P<0.05)。与DOX组对比,不同剂量GV1001组小鼠LVIDd、LVEF和LVFS升高,心肌损伤减轻,空泡化减轻,胶原沉积下降,血清cTnI、CK-MB和LDH下降,心肌组织MDA下降,T-SOD和GSH-PX升高(均P<0.05)。与对照组比较,DOX组小鼠心脏组织中cGAS、p-STING和p-IRF3的蛋白表达量均升高(P<0.05);DOX小鼠经GV1001治疗后cGAS、p-STING和p-IRF3的表达下降(P<0.05)。使用STING激动剂agonist干预后,GV1001对DOX小鼠心功能的改善作用减弱,对心脏组织氧化应激的抑制减轻。结论:GV1001可以缓解DOX诱导的小鼠心肌损伤,其机制可能是通过抑制cGAS/STING通路减轻氧化应激水平。

柚皮素抑制cGAS-STING通路活化对大鼠下肢深静脉血栓形成的影响

【目的】探讨柚皮素抗大鼠下肢深静脉血栓形成机制。【方法】将60只大鼠随机分为6组,即假手术组,模型组,柚皮素低、中、高剂量组和柚皮素高剂量+STING激动剂2.5己酮可可碱(DMXAA)组,每组10只。检测各组大鼠凝血指标[D-二聚体(D-dimer)、凝血酶时间(TT)、活化部分凝血活酶时间(APTT)、凝血酶原时间(PT)],炎症指标[白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)]及氧化应激指标[丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)];苏木素-伊红(HE)染色法检测静脉血栓形成;检测血栓湿质量和干质量;透射电镜检测静脉血栓超微结构;Western Blot法检测静脉血栓组织中环鸟苷酸-腺苷酸合成酶(c GAS)、干扰素基因刺激因子(STING)蛋白表达。【结果】(1)与假手术组比较,模型组大鼠D-dimer水平,IL-1β、IL-6、TNF-α水平,MDA含量,血栓湿质量和干质量增加,TT、APTT、PT,SOD活性和GSH-Px活性降低(均P<0.05);与模型组比较,柚皮素低、中、高剂量组大鼠D-dimer水平,IL-1β、IL-6、TNF-α水平,MDA含量,血栓湿质量和干质量降低,TT、APTT、PT,SOD活性和GSH-Px活性增加(均P<0.05),且呈剂量依赖性;与柚皮素高剂量组比较,柚皮素高剂量+DMXAA组大鼠D-dimer水平,IL-1β、IL-6、TNF-α水平,MDA含量,血栓湿质量和干质量升高,TT、APTT、PT,SOD活性和GSH-Px活性降低(均P<0.05)。(2)与假手术组比较,模型组大鼠静脉血栓组织中cGAS、STING蛋白表达水平升高(P<0.05);与模型组比较,柚皮素低、中、高剂量组大鼠静脉血栓组织中cGAS、STING蛋白表达水平均显著降低(P<0.05);柚皮素高剂量+DMXAA组大鼠cGAS、STING蛋白表达水平显著高于柚皮素高剂量组(P<0.05)。【结论】柚皮素可通过抑制cGAS/STING信号通路活化,抑制炎症、氧化应激反应,从而缓解下肢深静脉血栓形成。

STING高表达通过调控TLR4/NF-κB/NLRP3通路和影响炎症与凋亡水平促进小鼠肾脏缺血再灌注损伤

目的探讨STING在肾缺血再灌注损伤(IRI)中的表达水平以及相关作用机制。方法在体内水平,将24只C57BL/6小鼠分为假手术组(Sham)、IRI组、IRI+药物溶剂组(IRI+DMSO)、IRI+SN-011组,6只/组。通过肾动脉夹闭方法建立IRI模型,通过血清肌酐和尿素氮检测、PAS染色检测肾组织损伤变化,采用RT-qPCR、ELISA、Western blotting和IHC法检测肾组织中STING、KIM-1、Bcl-2、Bax、caspase-3、TLR4、P65、NLRP3、caspase-1、CD68、MPO、IL-1β、IL-6、TNF-α的水平。在体外水平,将HK-2细胞分为对照组、缺氧复氧(H/R)组、H/R+药物溶剂组(H/R+DMSO)、H/R+SN-011组,用厌氧包模拟缺氧环境,RT-qPCR和Western blotting法检测STING表达水平,流式细胞术检测各组细胞凋亡率。结果在体内水平,与Sham组相比,IRI组的PAS染色显示组织损伤增加(P<0.05),小鼠血清肌酐、尿素氮含量以及组织KIM-1、STING、TLR4、P65、NLRP3、caspase-1、caspase-3、Bax、CD68、MPO、IL-1β、IL-6、TNF-α表达水平升高(P<0.05),Bcl-2水平降低(P<0.05),SN-011抑制STING表达后,逆转了上述结果(P<0.05)。在体外水平,与对照组相比,H/R组STING的mRNA与蛋白水平升高(P<0.05),流式细胞仪检测显示细胞凋亡率上升(P<0.05),SN-011抑制STING表达,细胞凋亡率下降(P<0.05)。结论STING在肾脏IRI中表达水平上升,且可通过作用于TLR4/NF-κB/NLRP3通路以及影响炎症与凋亡水平促进肾损伤。

高良姜素通过激活cGAS/STING信号通路抑制骨肉瘤MG63细胞的恶性生物学行为

目的:探讨高良姜素(Gal)是否通过调节cGAS/STING信号通路影响骨肉瘤MG63细胞增殖、迁移、侵袭和凋亡。方法:体外培养人骨肉瘤MG63细胞,分别使用0、5、15、25、50、100、200μmol/L的Gal培养48 h后,CCK-8法检测Gal对细胞活力的影响。将MG63细胞分为对照组(未处理细胞)、Gal低浓度组(Gal-L组,50μmol/L Gal处理)、Gal高浓度组(Gal-H组,100μmol/L Gal处理)和Gal-H+STING抑制剂组(Gal-H+H-151组,100μmol/L Gal+8μmol/L H-151处理)。采用EdU染色法、划痕愈合实验、Transwell小室法、流式细胞术检测各组细胞增殖活力、迁移、侵袭和凋亡能力,WB法检测各组细胞中cGAS、STING蛋白的表达水平。结果:与对照组比较,Gal-L组、Gal-H组细胞增殖活力、迁移、侵袭能力均显著降低(均P<0.05),细胞凋亡率和cGAS、STING蛋白表达水平均显著升高(均P<0.05);与Gal-H组比较,Gal-H+H-151组细胞增殖活力、迁移和侵袭能力均显著升高(均P<0.05),细胞凋亡率和cGAS、STING蛋白表达水平均显著降低(均P<0.05)。结论:Gal可能通过激活cGAS/STING信号通路抑制骨肉瘤MG63细胞增殖、迁移、侵袭并促进细胞凋亡。

现代与传承:STING激动剂的双重视角

cGAS-STING信号通路是免疫系统中的关键调节因子,对肿瘤免疫治疗具有重要的研究和应用前景。该信号通路激活STING蛋白后,能够促进I型干扰素(IFN-I)的产生,并进一步影响树突细胞的成熟,增强T淋巴细胞和自然杀伤(NK)细胞的攻击力,从而连接先天免疫和适应性免疫反应。文章重点回顾了化学类STING激动剂以及中药成分对cGAS-STING信号通路的调节作用。这些研究不仅揭示了STING激动剂的分子机制,而且为开发新型免疫调节剂提供了理论基础。此外,文章还探讨了中药成分与现代药物设计的结合潜力,为未来免疫治疗的研究提供了新的视角和策略。