Huatan Qushi formula alleviates non-alcoholic fatty liver disease via PI3K/Akt signaling and gut microbiota modulation

Objective:To provide the mechanism-based pharmacotherapy of the Huatan Qushi formula(HTQS for-mula),for the health management and treatment of non-alcoholic fatty liver disease(NAFLD).Methods:A rat model of NAFLD was employed to examine the efficacy and safety of the HTQS formula.In vivo active components and potential mechanisms of the HTQS formula were identified using UPLC‒MS/MS combined with network pharmacology.The influence of the HTQS formula on the dominating proteins in PI3K/Akt pathway was validated in vivo using western blot.Finally,16S rRNA sequencing of the gut microbiome was conducted followed by targeted metabolomics detecting fecal short-chain fatty acids(SCFAs)and bile acids to determine the impact of the HTQS formula on gut microbiota.Results:The HTQS formula reduced weight gain and hepatic steatosis in NAFLD rats and decreased serum total cholesterol(TC),triglycerides,blood glucose,and insulin resistance(IR)without causing liver or kidney injury.We detected 28 components using UPLC‒MS/MS and identified 439 shared targets be-tween NAFLD and the HTQS formula.Primarily,we focused on the PI3K/Akt signaling pathway based on protein‒protein interaction network analysis.We validated that the HTQS formula inhibited liver stea-tosis and inflammation by increasing the phosphorylation levels of PI3K,AKT,P27,GSK3b in the PI3K/Akt signaling pathway.16S rRNA sequencing revealed that the HTQS formula reduced the abundance of the genus Family_XIII_AD3011_group,which was positively correlated with IR and taurodeoxycholic acid.In addition,Lachnospiraceae_UCG_010 inversely correlated with TC and five bile acids,which could be essential to the therapeutic effect of the HTQS formula against NAFLD.Conclusions:The HTQS formula proved to be an effective pharmacotherapy for NAFLD without causing liver or kidney injury.Multiple potent components of the HTQS formula could alleviate liver steatosis and lipid metabolism disorder by modulating the PI3K/Akt signaling pathway and restoring gut microbiota composition.

中药Formula-3抑制RBL-2H3细胞脱颗粒机制的研究

目的通过研究中药复方Formula-3(乌梅、灵芝、人参、当归、黄连、干姜)对嗜碱性粒细胞系(RBL-2H3细胞)和SOC通道的作用,探讨Formula-3对肥大细胞脱颗粒的作用机制。方法体外培养大鼠RBL-2H3细胞系制作肥大细胞的模型,通过MTT实验、脱颗粒实验和激光共聚焦(Confocal)实验来研究Formula-3对肥大细胞脱颗粒的作用机制。结果 Formula-3(终质量浓度0.05、0.2、0.8 mg/ml)在急性(0.5 h)和慢性(12 h)作用下呈质量浓度依赖性的抑制致敏RBL-2H3细胞脱颗粒(P<0.01),而对非致敏RBL-2H3细胞没有影响(P>0.05)。Formula-3可显著的抑制SOC通道钙离子内流(P<0.01)。结论 Formula-3通过抑制SOC通道钙离子内流来显著抑制肥大细胞脱颗粒,为治疗过敏性疾病提供理论依据。

中药Formula-3治疗BN大鼠食物过敏的研究

目的探讨中药Formula-3治疗BN大鼠食物过敏的效果及作用机制。方法建立卵清蛋白(OVA)大鼠食物过敏模型,通过中药Formula-3治疗后对相关指标进行检测。结果与OVA实验组相比,中药Formula-3治疗后血清中特异性Ig E水平明显降低(P<0.05),血清中组胺水平亦降低(P<0.05)。肠组织切片HE染色和甲苯胺蓝染色可见Formula-3治疗组肠组织病理变化和肥大细胞脱颗粒现象较OVA实验组有了显著的改善(P<0.05)。结论中药Formula-3可以有效治疗BN大鼠食物过敏,为治疗食物过敏性疾病提供理论依据。

镁掺杂对ZnO-Al_2O_3系陶瓷性能的影响

运用回归数学分析并结合ＳＥＭ观测结果，对影响ＺｎＯ－Ａｌ２Ｏ３系陶瓷电阻及阻－温特性的线性化机制进行了探讨。研究表明，Ａｌ２Ｏ３、ＭｇＯ的掺杂及烧成工艺均对材料的电阻率和电阻温度系数有较为明显的影响，镁掺杂对材料的电子激活能有较大的影响，当材料的电子激活能值较低时，通过回归处理发现其阻－温特性具有较好的线性，符合麦克劳林公式。

痛风合剂3号方治疗慢性尿酸性肾病脾肾亏虚证的临床效果

目的:探讨慢性尿酸性肾病(CUAN)脾肾亏虚证患者采用痛风合剂3号方治疗的临床效果。方法:选取2017年1月—2022年12月泉州市中医院收治的60例CUAN脾肾亏虚证患者作为研究对象,根据随机数表法分为研究组和对照组,每组各30例。对照组给予常规西药治疗,研究组给予痛风合剂3号方治疗。比较两组临床疗效、中医症候积分、肾功能、炎症反应及不良反应情况。结果:研究组治疗总有效率较对照组高,差异有统计学意义(P<0.05);治疗后,两组中医症候积分均下降,研究组较对照组更低,差异有统计学意义(P<0.05);治疗后,两组尿素氮(BUN)、24 h尿蛋白定量(24 hUpro)、中性粒细胞明胶酶相关脂质运载蛋白(NGAL)均下降,肾小球滤过率(eGFR)升高,研究组BUN、24hUpro、NGAL较对照组更低,eGFR更高,差异有统计学意义(P<0.05);治疗后,两组白细胞介素-6(IL-6)、C反应蛋白(CRP)水平下降,研究组较对照组更低,差异有统计学意义(P<0.05);两组不良反应发生率比较,差异无统计学意义(P>0.05)。结论:CUAN脾肾亏虚证患者采用痛风合剂3号方治疗,能够提高临床疗效,缓解中医症候,改善肾功能的同时降低炎症反应,且不会增加不良反应。

培本清利通络方治疗慢性肾脏病3~5期伴CKD-MBD脾肾两虚兼湿瘀证的临床研究

目的:观察基于吴门医派“络病理论”创立的培本清利通络方联合骨化三醇胶丸治疗慢性肾脏病(CKD)3~5期伴CKD-矿物质及骨代谢紊乱(CKD-MBD)脾肾两虚兼湿瘀证患者的效果。方法:选择2022年7月—2023年6月于南京中医药大学附属苏州市中医医院就诊的脾肾两虚兼湿瘀证CKD 3~5期合并有CKD-MBD的患者60例,随机分为治疗组和对照组,每组30例。对照组予控制血压、控制血糖、改善贫血等基础治疗,同时口服骨化三醇胶丸;治疗组在对照组治疗基础上加服培本清利通络方,两组疗程均为12周。比较两组患者治疗前后肾功能[血肌酐(Scr)、血尿素氮(BUN)、尿酸(UA)]、矿物质及骨代谢[钙(Ca)、磷(P)、全段甲状旁腺激素(iPTH)、碱性磷酸酶(ALP)]、中医症候积分、生活质量评分,并评估临床疗效;治疗前后检测两组血常规、肝功能、血钾,以评估用药安全性。结果:两组治疗后中医症候积分均较治疗前降低,且治疗组低于对照组(P<0.05);治疗组总有效率为93.33%,明显高于对照组的73.33%(P<0.05);治疗后,治疗组BUN、UA均明显低于治疗前,且Scr、BUN均明显低于对照组,差异均有统计学意义(P<0.05);治疗后,治疗组P、iPTH均明显低于治疗前,Ca明显高于治疗前,且治疗组Ca高于对照组,iPTH低于对照组,差异均有统计学意义(P<0.05);治疗后,治疗组生活质量评分较治疗前明显下降,且治疗组低于对照组(P<0.05);两组治疗后安全性指标比较,差异均无统计学意义(P>0.05)。结论:培本清利通络方可改善CKD 3~5期合并CKD-MBD脾肾两虚兼湿瘀证患者Ca、P、iPTH指标,延缓肾功能减退,减轻患者腰脊酸痛、皮肤瘙痒、倦怠乏力等症状,并可提高患者生活质量。

Guizhi-Fuling formula inhibits ovarian cancer progression by targeting STAT3 signaling network

Objective:Ovarian cancer(OC)is the most lethal gynecological malignancy.Frequent peritoneal dissemination is the main cause of low survival rate.Guizhi-Fuling formula(GZFL)is a classical traditional Chinese herbal formula,and has been clinically used for treating ovarian cancer with good outcome.However,its therapeutic mechanism for treating OC has not been clearly elucidated.Methods:Network pharmacology analysis was used to predict potential molecular mechanisms of GZFL in treating OC.In vitro and in vivo analysis,including STAT3 KO/WT cells proliferation assay,scratch assay and antitumor efficacy study were performed to assess the biological activity of GZFL on targeting STAT3 in OC cells.Results:We generated a“GZFL target-OC-STAT3”gene interaction network,and predicted that GZFL is tightly associated with IL6/JAK/STAT3 signal pathway and cancer metastasis.Our preliminary data showed that GZFL inhibited OC cell proliferation in a STAT3 dependent manner.It suppressed cell migration and downregulated p-STAT3 expression.In a tumor bearing mouse model,GZFL displayed a safety profile.Conclusion:GZFL inhibits OC progression by targeting STAT3 signaling network.Our newly proposed pharmacological mechanisms of Guizhi-Fuling formula will provide a new insight for its clinical use in treating OC.

橘皮竹茹汤对化疗性异食癖大鼠模型5-羟色胺3受体蛋白和mRNA表达的影响

目的观察橘皮竹茹汤对化疗性异食癖大鼠模型止呕作用及对5-羟色胺3受体(5-HT3R)蛋白和mRNA的影响。方法将36只Wistar雄性大鼠随机分为正常对照组、中药对照组(橘皮竹茹汤10.6 g/kg,不造模)、模型组、昂丹司琼组(2.6 mg/kg)、橘皮竹茹汤低剂量组(5.3 g/kg)和橘皮竹茹汤高剂量组(10.6 g/kg),每组6只。各组大鼠给予相应干预措施6 d(每日2次)后,采用腹腔注射顺铂(6 mg/kg)的方式诱导建立化疗性异食癖大鼠模型,观察并记录造模后24 h内大鼠体质量、摄食量和高岭土摄入量的变化。继续干预1 d后,苏木精-伊红(HE)染色观察各组大鼠胃窦、回肠组织形态学变化;实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法检测延髓、回肠组织中5-HT3R、色氨酸羟化酶(TPH)、单胺氧化酶A(MAOA)mRNA表达;Western blot法检测延髓组织中5-HT3R蛋白的表达。结果①与正常对照组比较,模型组大鼠体质量、摄食量降低(P<0.05),高岭土摄入量显著增加(P<0.05);与模型组比较,昂丹司琼组和橘皮竹茹汤低、高剂量组大鼠高岭土摄入量显著降低(P<0.05)。②HE染色显示,与正常对照组比较,模型组胃窦组织上皮细胞受损,固有层腺体排列疏松、紊乱,回肠上皮部分缺失,腺体排列紊乱;与模型组比较,昂丹司琼组和橘皮竹茹汤低、高剂组胃窦和回肠组织病理改变减轻。③RT-qPCR结果显示,与正常对照组比较,模型组大鼠延髓、回肠5-HT3R mRNA表达水平显著升高(P<0.05),回肠MAOA mRNA表达水平显著降低(P<0.05);与模型组比较,橘皮竹茹汤高剂量组大鼠延髓5-HT3R mRNA表达水平明显降低(P<0.05),橘皮竹茹汤低剂量组回肠TPH1 mRNA、橘皮竹茹汤高剂量组延髓TPH2 mRNA表达水平明显降低(P<0.05),昂丹司琼组、橘皮竹茹汤低剂量组、橘皮竹茹汤高剂量组回肠MAOA mRNA表达水平明显升高(P<0.05)。④Western blot结果显示,与正常对照组比较,模型组大鼠延髓5⁃HT3R蛋白表达水平显著升高(P<0.05);与模型组比较,橘皮竹茹汤高剂量组大鼠延髓5-HT3R蛋白表达水平显著降低(P<0.05)。结论橘皮竹茹汤可以通过抑制延髓5-HT3R蛋白和mRNA的表达,调控5-羟色胺合成与代谢相关酶,进而改善化疗导致的恶心呕吐。

Microwave absorption properties regulation and bandwidth formula of oriented Y_(2)Fe_(17)N_(3-δ)@SiO_(2)/PU composite synthesized by reduction-diffusion method

As concepts closely related to microwave absorption properties,impedance matching and phase matching were rarely combined with material parameters to regulate properties and explore related mechanisms.In this work,reduction–diffusion method was innovatively applied to synthesize rare earth alloy Y_(2)Fe_(17).In order to regulate the electromagnetic parameters of absorbers,the Y_(2)Fe_(17)N_(3-δ)particles were coated with silica(Y_(2)Fe_(17)N_(3-δ)@SiO_(2))and absorbers with different volume fractions were prepared.The relationship between impedance matching,matching thickness,and the strongest reflection loss peak(RLmin)was presented obviously.Compared to the microwave absorption properties of Y_(2)Fe_(17)N_(3-δ)/PU absorber,Y_(2)Fe_(17)N_(3-δ)@SiO_(2)/PU absorbers are more conducive to the realization of microwave absorption material standards which are thin thickness,light weight,strong absorbing intensity,and broad bandwidth.Based on microwave frequency bands,the microwave absorption properties of the absorbers were analyzed and the related parameters were listed.As an important parameter related to perfect matching,reflection factor(√ε_(r)/μ_(r))was discussed combined with microwave amplitude attenuation.According to the origin and mathematical model of bandwidth,the formula of EAB(RL<-10 dB)was derived and simplified.The calculated bandwidths agreed well with experimental results.

To investigate the effect of Shenqi Tiaoshen Formula on CSE induced inflammatory response of MH-S cells based on TLR4/NF-kB/NLRP3 pathway

Objective:To study the effects of Shenqi Tiaoshen Formula(SQTS)on the inflammatory response of MH-S cells induced by cigarette smoking extract(CSE)and its mechanism based on TLR4/NF-kB/NLRP3 pathway.Methods:MH-S cells were used as subjects to evaluate cell viability by CCK-8 method.The levels of TNF-α,IL-1βand IL-6 in the supernatant were detected by ELISA.ROS were detected by DCFH-DA fluorescence probe.Western blotting was used to detect the expression of TLR4/NF-kB/NLRP3 pathway protein,and TAK-242,a TLR4 inhibitor,was used to verify the role of SQTS in the TLR4/NF-kB/NLRP3 pathway.Results:Compared with blank group,the cell survival rate of CSE group was decreased,and the contents of inflammatory cytokines TNF-α,IL-1βand IL-6 were increased(P<0.05),ROS fluorescence expression level was significantly increased(P<0.01),TLR4/NF-kB/NLRP3 pathway protein expression was significantly increased(P<0.05);Compared with CSE group,the survival rate of cells in SQTS groups was increased,and the expression levels of the above indexes were decreased(P<0.05),and TLR4/NF-kB/NLRP3 pathway protein decreased in TAK-242 groups(P<0.05).Conclusion:SQTS can reduce the inflammatory response of MH-S cells induced by CSE by inhibiting TLR4/NF-kB/NLRP3 pathway.

To explore the mechanism of Dahuang Lingxian Formula in relieving inflammatory response of bile duct cells based on IL-6/JAK/STAT3 signaling pathway

Objective:To explore the mechanism of action of Dahuang Lingxian Formula in alleviating the inflammatory response of bile duct cells in LPS-induced intrahepatic bile duct inflammation model rats based on IL-6/JAK/STAT3 signaling pathway.Methods:Fifty SD rats were randomly divided into five groups,blank group,model group,choling tablets(0.5 g/kg),and low and high concentration groups(2.4 g/kg and 4.8 g/kg)of Dahuang Lingxian Formula,ten rats in each group.Except for the blank group,the rats in each group were injected with 1.25 mg/kg LPS at the common bile duct at one time to construct an animal model of intrahepatic bile duct infection.After gavage on day 8,liver tissues were taken from rats at the hepatic hilum,and the histopathological changes of the hepatic hilum and biliary tree were observed by HE staining.The expression levels of serum glutamic alanine transaminase(ALT),glutamic oxalacetic transaminase(AST),malondialdehyde(MDA)and superoxide dismutase(SOD)were measured by biochemical method.The expression levels of interleukin 6(IL-6),Janus protein tyrosine kinase 2(JAK2),signal transducer and activator of transcription 3(STAT3)in rat serum were measured by enzyme-linked immunosorbent assay(ELISA).Protein immunoblotting(WB)and real-time fluorescence quantitative PCR(RT-qPCR)were used to detect the expression levels of IL-6,JAK2,STAT3 protein and mRNA in biliary tree tissues.Results:①Compared with the blank group,the structures such as interlobular bile ducts in the hepatic sinusoids and portal duct area of the model rats were destroyed,and inflammatory cells infiltrated around them.The expression of ALT,AST,MDA,IL-6,JAK2 and STAT3 in the serum increased significantly,the expression level of SOD decreased,and the expression levels of IL-6,JAK2 and STAT3 proteins and mRNA increased.②Compared with the model group,the degree of liver pathological damage in rats in the Chiling Ning tablet group and the low and high concentration groups of Dahuang Lingxian Formula were improved,which could significantly reduce the expression levels of ALT,AST,MDA,IL-6,JAK2,STAT3 and up-regulate SOD in serum,and down-regulate the expression of IL-6,JAK2,STAT3 protein and mRNA,with the best effect in the high concentration group of Dahuang Lingxian Formula.③Compared with the choling tablet group,the rats in the low and high concentration groups of Dahuang Lingxian Formula tended to normalize the degree of liver pathological damage,without obvious inflammatory cell infiltration,and the expression levels of ALT,AST,MDA,IL-6,JAK2,STAT3 and the expression levels of IL-6,JAK2,STAT3 protein and mRNA in serum were reduced,and the expression levels of SOD were increased,with the best effect of Dahuang Lingxian Formula The treatment effect was best in the high concentration group.Conclusion:The mechanism may be related to the down-regulation of IL-6/JAK/STAT3 signaling pathway activation,and the best therapeutic effect was achieved by the high concentration group of Dahuang Lingxian Formula.

Effects of Cigu Xiaozhi Formula on miR-378a-3p Expression and Hh Signaling Pathway in TGF-β1 Induced LX2 Cells

[Objectives]To observe the effects of Cigu Xiaozhi Formula on miR-378a-3p expression and Hh signaling pathway in TGF-β1 induced and activated LX2 cells.[Methods]Cells were divided into control group,induction group,drug-containing serum group,miR-378a-3p inhibitor group,and miR inhibitor NC group.CCK-8 method was used to detect the cell viability of each group,and flow cytometry was used to detect the apoptosis rate of each group.RT-qPCR was used to detect the expression of miR-378a-3p in each group s cells,and RT-qPCR and Western blot were used to detect mRNA and protein expression of Shh,Gli1,Gli2,Col-I,andα-SMA in each group s cells.[Results]Compared with the control group,the cell viability and expression of Shh,Gli1,Gli2,Col-I,andα-SMA mRNA and protein in induction group increased(P<0.01),while the expression of miR-378a-3p decreased(P<0.01).Compared with the induction group,the cell viability and expression of Shh,Gli1,Gli2,Col-I,α-SMA mRNA andα-SMA and Gli2 protein decreased in drug-containing serum group(P<0.05),while cell apoptosis rate and miR-378a-3p expression increased(P<0.01).In miR-378a-3p inhibitor group,cell viability and the expression of Shh,Gli1,Gli2,Col-I,α-SMA mRNA and Gli1,Gli2,α-SMA protein increased(P<0.05,P<0.01),while the apoptosis rate and miR-378a-3p expression decreased(P<0.05,P<0.01).[Conclusions]Cigu Xiaozhi Formula containing serum can upregulate miR-378a-3p expression and downregulate the expression of Gli2 andα-SMA in TGF-β1 induced LX2 cells,thereby inhibiting the activation of LX2 cells and exerting the effects of anti liver fibrosis.

保肺化纤方调控TGF-β1/smad2/3、Wnt1/β-catenin信号通路对BLM/PM2.5诱导的特发性肺纤维化大鼠肺组织胶原沉积的影响

目的探讨保肺化纤方经转化生长因子(transforming growth factor,TGF)-β1/smad2/3、Wnt1/β-连环蛋白(catenin)信号通路减轻博来霉素(bleomycin,BLM)/PM2.5暴露诱导的特发性肺纤维化(idiopathic pulmonary fibrosis,IPF)大鼠肺组织胶原沉积的作用机制。方法SD大鼠随机分为对照(Control)组、BLM+PM2.5(Model)组、保肺化纤方(BHF)组、吡非尼酮(pirfenidone,PFD)组、XAV-939组、BHF+PFD组、BHF+XAV-939组。采用一次性气管内雾化BLM法制备IPF大鼠模型,大气PM2.5实时浓缩全身暴露和给予相应药物干预。检测大鼠肺功能,观察肺组织病理和胶原沉积;采用免疫组化技术检测肺组织Ⅰ、Ⅳ型胶原(collagen,COL)、TGF-β1、smad2/3、β-catenin蛋白水平;采用qPCR技术检测肺组织TGF-β1、smad2/3、Wnt1、β-catenin mRNA水平。结果与Control组比较,Model组大鼠肺功能显著降低(P<0.01),肺组织可见大量炎症细胞浸润,肺泡壁断裂、增厚,胶原沉积等肺纤维化特征性病理改变,肺组织胶原容积分数(collagen volume fraction,CVF)、COLⅠ、COLⅣ蛋白及TGF-β1、smad2/3、β-catenin基因与蛋白表达均显著升高(P<0.01);与Model组比较,各治疗组均有效改善大鼠肺组织病理改变,显著降低肺组织CVF及COLⅠ、COLⅣ、TGF-β1、smad2/3、β-catenin蛋白表达水平(P<0.05,P<0.01),同时BHF组、BHF+PFD组、BHF+XAV-939组均能显著提高大鼠肺功能(P<0.05,P<0.01),明显下调肺组织TGF-β1、smad2/3、β-catenin基因表达水平(P<0.01);与PFD组比较,BHF+PFD组大鼠肺组织COLⅠ、TGF-β1蛋白及smad2、β-catenin基因表达均显著降低(P<0.05,P<0.01);与XAV-939组比较,BHF+XAV-939组smad3基因表达显著降低(P<0.01)。结论保肺化纤方可改善BLM/PM2.5诱导的IPF大鼠肺组织病理损伤,减少胶原沉积,提高肺功能,改善肺纤维化,其机制可能与下调TGF-β1/smad2/3、Wnt1/β-catenin通路有关。

保肾排毒方联合乌蕨汤保留灌肠对慢性肾脏病3~5期患者的临床疗效

目的 探讨保肾排毒方联合乌蕨汤保留灌肠对慢性肾脏病3~5期患者的临床疗效。方法 80例患者随机分为对照组和观察组,每组40例,对照组给予常规治疗,观察组在对照组基础上加用保肾排毒方联合乌蕨汤保留灌肠,疗程8周。检测临床疗效、炎性因子(CRP、IL-6、TNF-α)、肾功能指标(BUN、Scr、β2-MG、PTH)、中医证候评分、不良反应发生率变化。结果 观察组总有效率高于对照组(P<0.05)。治疗后,2组炎性因子、肾功能指标、中医证候评分降低(P<0.05),以观察组更明显(P<0.05)。2组不良反应发生率比较,差异无统计学意义(P>0.05)。结论 保肾排毒方联合乌蕨汤保留灌肠可安全有效地改善慢性肾脏病3~5期患者肾功能,减轻中医证候,抑制炎症反应。

祛风止痒方超声雾化对过敏性结膜炎模型兔炎症因子IL-3、IL-5、GM-CSF、TSLP及相关蛋白表达的影响

目的观察祛风止痒方超声雾化对过敏性结膜炎(allergic conjunctivitis,AC)模型兔结膜组织中Janus酪氨酸激酶2/信号转导子和转录激活子5(Janus kinase2/signal transducer and activator of transcription5,JAK2/STAT5)信号转导通路关键细胞因子白细胞介素-3(interleukin-3,IL-3)、白细胞介素-5(interleukin-5,IL-5)、粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony-stimulating factor,GM-CSF)及胸腺基质淋巴细胞生成素(thymic stromal lymphopoietin,TSLP)表达的影响。方法选取健康成年新西兰长耳大白兔36只,按照随机数字表法分为对照组、模型组、奥洛他定组、祛风止痒组,每组9只。对照组不予处理,其余组分别于实验第0、7天予静脉注射含卵清蛋白(ovalbumin,OVA)和佐剂氢氧化铝的混合物致敏,此后每天采用OVA、磷酸盐平衡生理盐水混合物局部每天滴眼1次,连续14 d,以加强致敏,建立AC模型。造模成功后,奥洛他定组用盐酸奥洛他定滴眼液滴双眼,祛风止痒组用祛风止痒方超声雾化治疗,连续给药14 d。造模期间裂隙灯下观察兔眼前节临床表现,并采用Magone计分法行眼部症状临床评分。末次给药24 h后,以摘眼球法采血并取兔睑结膜组织,ELISA检测血清IL-3、IL-5、GM-CSF及结膜组织TSLP含量,Western blot检测结膜组织JAK2、STAT5蛋白表达水平。结果模型组Magone评分明显高于对照组(P<0.05),说明AC模型制备成功。与对照组比较,模型组兔眼部可见明显的结膜充血、分泌物及流泪表现;与模型组比较,奥洛他定组及祛风止痒组兔眼部结膜仅见少量充血、分泌物,流泪症状减轻。与对照组比较,模型组血清IL-3、IL-5、GM-CSF及结膜组织TSLP含量均升高(P<0.05),结膜组织中JAK2、STAT5蛋白表达水平均升高(P<0.05);与模型组比较,奥洛他定组及祛风止痒组血清IL-3、IL-5、GM-CSF及结膜组织TSLP含量均降低(P<0.05),结膜组织中JAK2、STAT5蛋白表达水平均下降(P<0.05);与奥洛他定组比较,祛风止痒组血清IL-3、IL-5、GM-CSF及结膜组织TSLP含量差异均无统计学意义(P>0.05),结膜组织中JAK2、STAT5蛋白表达水平均升高(P<0.05)。结论祛风止痒方超声雾化对OVA诱导的AC样症状有明显疗效,其作用机制可能与抑制JAK2/STAT5信号转导通路中关键细胞因子,进而减轻炎症反应有关。

基于JAK2/STAT3通路探讨活血荣络方对OGD/R后BMEC的干预作用及机制

目的探讨活血荣络方激活Janus酪氨酸激酶2/信号转导和转录激活剂3(Janus kinase 2/signal transducer and activator of transcription 3,JAK2/STAT3)通路对氧糖剥夺/复氧(oxygen-glucose deprivation/reperfusion,OGD/R)后脑微血管内皮细胞(brain microvascular endothelial cell,BMEC)的干预作用和机制。方法培养大鼠BMEC(bEnd.3细胞),建立OGD/R模型;将细胞随机分为正常组(10%空白血清)、模型组(10%空白血清)、活血荣络方组(10%活血荣络方含药血清)、丁苯酞组(10%丁苯酞含药血清)、Stattic组(10%空白血清+10μmol/mL Stattic)、联用组(10%活血荣络方含药血清+10μmol/mL Stattic),并分别给予相应的药物干预24 h。采用CCK-8法检测活血荣络方对OGD/R后bEnd.3细胞活性的影响;采用划痕实验和Transwell迁移实验分别检测活血荣络方对OGD/R后bEnd.3细胞划痕愈合和细胞迁移的影响;采用Western blot法检测bEnd.3细胞中JAK2、p-JAK2、STAT3、p-STAT3、血管内皮细胞生长因子A(vascular endothelial growth factor A,VEGFA)蛋白表达水平。结果与正常组比较,模型组bEnd.3细胞存活率、细胞划痕愈合率、细胞迁移数显著降低(P<0.01),JAK2、p-JAK2、STAT3、p-STAT3、VEGFA的表达量升高(P<0.05或P<0.01);与模型组比较,活血荣络方组bEnd.3细胞的存活率、细胞划痕愈合率、细胞迁移数显著升高(P<0.01),JAK2、p-JAK2、p-STAT3、STAT3、VEGFA的表达量升高(P<0.05或P<0.01);与Stattic组比较,联用组bEnd.3细胞的存活率、细胞划痕愈合率、细胞迁移数显著升高(P<0.05或P<0.01),STAT3、VEGFA的表达量升高(P<0.05)。结论活血荣络方可能激活JAK2/STAT3信号通路并诱导内皮细胞的增殖、迁移和存活,激活VEGFA表达并减轻脑缺血再灌注损伤,具有神经保护作用。

海珠益肝加味方对自身免疫性肝炎小鼠Th17/Treg平衡与FoxP3/RORγt的影响

目的:研究海珠益肝加味方对自身免疫性肝炎(AIH)小鼠辅助性T淋巴细胞17(Th17)/调节性T细胞(Treg)细胞失衡与维甲酸相关孤核受体γ(ROR-γt)、特异性叉头状转录因子3(FoxP3)的影响。方法:将32只小鼠随机分为正常组、模型组、海珠益肝加味方组、醋酸泼尼松组,每组8只。除正常组外其他组采用刀豆蛋白A(ConA)尾静脉注射制备AIH小鼠模型,造模后采集各组小鼠血液及肝组织。HE染色观察各组肝组织病理表现;采用流式细胞术检测各组Th17和Treg细胞比例。Western Blot检测各组小鼠肝组织中IL-10、IL-17、TGF-β1、FoxP3、ROR-γt蛋白表达水平。结果:与空白组比较,模型组Th17细胞比例、Th17/Treg均显著上升(P<0.01),Treg细胞比例显著下降(P<0.01);与模型组比较,海珠益肝加味方组及醋酸泼尼松组Th17细胞比例、Th17/Treg均显著降低(P<0.01),Treg细胞比例显著上升(P<0.01)。与空白组比较,模型组IL-10、FoxP3蛋白表达下降,IL-17、TGF-β1、ROR-γt蛋白表达上升(P<0.01)。与模型组相比,海珠益肝加味方及醋酸泼尼松组IL-10、FoxP3表达显著上升,IL-17、TGF-β1、RORγt蛋白表达均显著下降(P<0.01)。结论:海珠益肝加味方可以有效改善Th17/Treg失衡,其机制可能与调节特异性转录因子FoxP3、ROR-γt的表达相关。

抗炎通腑方对脓毒症急性肺损伤大鼠mtDNA-STING-NLRP3信号通路的影响

目的探讨抗炎通腑方对脓毒症急性肺损伤(ALI)大鼠的影响及作用机制。方法将35只大鼠随机分为正常组、模型组、地塞米松(DEX)组、中药(TCM)组和TCM+DEX组,每组7只。分别给予相应药物灌胃、造模。酶联免疫吸附实验(ELISA)法检测白细胞介素(IL)-6、IL-1β、IL-18、肿瘤坏死因子-α(TNF-α)含量及血清中巨噬细胞计数;采用实时荧光定量多聚核苷酸链式反应(RT-qPCR)和蛋白免疫印迹(Western blot)检测肺组织中干扰素基因刺激因子(STING)、磷酸化干扰素基因刺激因子(P-STING)、NOD样受体热蛋白结构域相关蛋白3(NLRP3)、凋亡相关斑点样蛋白信使核糖核酸(ASC mRNA)和相关蛋白的表达水平。测定肺组织湿/干比(W/D),观察肺组织病理学改变。检测肺组织中巨噬细胞表达情况。结果与正常组比较,各组指标显著升高;与模型组比较,DEX组、TCM组及TCM+DEX组各指标均显著降低。与模型组比较,TCM+DEX组改变最为明显,肺组织损伤改善,W/D值(4.77±0.29 vs.3.78±0.48)及巨噬细胞计数(13.39±2.06 vs.7.09±1.42)均降低(P<0.05),IL-6(152.51±22.27 vs.58.92±13.53)、IL-1β(126.19±10.02 vs.45.69±6.67)、IL-18(59.12±6.31 vs.31.75±4.23)及TNF-α(126.57±8.25 vs.49.59±8.12)水平均降低(P均<0.01),肺组织中线粒体DNA(mtDNA)损伤及释放、P-STING(0.32±0.03 vs.0.16±0.03)、STING mRNA(19.24±2.70 vs.0.32±0.16)、NLRP3 mRNA(20.03±5.06 vs.1.20±0.04)、ASC mRNA(16.96±4.31 vs.3.41±2.52)和相关蛋白的表达均降低(P<0.01)。结论抗炎通腑方可能通过调控mtDNA-STING-NLRP3信号通路减轻脓毒症ALI大鼠的炎症。

肾纤康通过调节Smad3介导的铁死亡缓解CKD小鼠肾纤维化

目的:观察肾纤康对单侧输尿管梗阻(unilateral ureteral obstruction,UUO)引起的小鼠肾间质纤维化的缓解效果,并探讨其作用机制。方法:将小鼠随机分为假手术组、模型组、低剂量(150 mg·kg^(−1)·d^(−1))肾纤康组和高剂量(450 mg·kg^(−1)·d^(−1))肾纤康组,每组8只。除假手术组外,其余各组均通过单侧输尿管结扎法建立慢性肾脏病(chronic kidney disease,CKD)模型。造模后,分别给予相应剂量的肾纤康,假手术组和模型组则灌胃等体积生理盐水,每天1次,连续7 d。实验结束后,收集小鼠肾脏组织进行苏木精-伊红(hematoxylin-eosin,HE)染色和Masson染色,观察肾脏组织损伤和纤维化程度;利用免疫组化和Western blot技术检测肾脏中纤维化、氧化应激及铁死亡相关蛋白水平的变化;蛋白质免疫共沉淀(co-immunoprecipitation,Co-IP)评估肾纤康对转录激活因子3(activating transcription factor 3,ATF3)与Smad3相互作用的影响。结果:在未经治疗的UUO模型组中,小鼠肾脏出现了如肾小管明显扩张和胶原沉积等典型的病理变化,肾纤康干预组病理改变程度和纤维化明显减轻(P<0.05)。在分子层面,肾纤康显著降低了UUO模型组小鼠中Smad3磷酸化水平,以及ATF3、4-羟基壬烯醛(4-hydroxynonenal,4-HNE)和NADPH氧化酶4(NADPH oxidase 4,NOX4)的异常表达,同时增加了谷胱甘肽过氧化酶4(glutathione peroxidase 4,GPX4)和溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)表达。Co-IP实验结果表明,肾纤康显著影响了ATF3与Smad3的相互作用。结论:肾纤康有效缓解了UUO引起的小鼠肾间质纤维化,其潜在机制可能涉及对ATF3/Smad3相互作用的调节,进而减轻氧化应激和铁死亡,从而缓解肾脏纤维化。这些发现为肾纤康的进一步研究和临床应用提供了重要的科学依据。

PI3K/Akt信号通路调控子宫内膜异位症及中药干预作用研究进展

子宫内膜异位症(endometriosis,EMs)是由于子宫内膜间质或者腺体异常定植到子宫腔被覆黏膜以外部位而导致的疾病。磷脂肌醇3激酶(phosphoinositide 3-kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)信号通路参与病灶血管形成、异位组织细胞自噬与凋亡和迁移侵袭等过程,是调控EMs病理特性的经典通路之一。中药多组分、多靶点、多途径的特性在治疗EMs进程中优势显著,部分中药有效成分及中药复方通过干预PI3K/Akt信号通路,进而达到抑制内膜异位组织进展、减轻疼痛和缓解纤维化病变的治疗作用。该文通过阐述PI3K/Akt信号通路关键靶点与EMs的联系,归纳并总结国内外近年来中药通过调控PI3K/Akt信号通路治疗EMs的研究现状,旨在为利用中药及复方治疗EMs提供新的视角和思路。