Antitumor and synergistic effect of Chinese medicine “Bushen huayu jiedu recipe” and chemotherapy on transplanted animal hepatocarcinoma

AIM： To investigate the antitumor and synergistic effect of Chinese medicine “Bushen huayu jiedu recipe” （recipe for invigorating the kidney, removing blood stasis and toxic substances） and chemotherapy on mice hepatocarcinoma. METHODS： Bushen huayu jiedu recipe （BSHYJDR） consisting of Chinese Cassia Bark, Psoralea, Zedoary, Rhubarb, etc. is equal to 1.5 g/mL liquid of originated herbs after being decoded, filtered, and concentrated. Kunming mice, weighing 18-22 g, were injected with 0.2 mL ascitic hepatocarcinoma H22 containing 1 × 10^7 cells/mL into armpit of the right forelimb of mice. After 24 h, the mice were weighed and randomly divided into tumor-bearing model control group, cisplatin （DDP） group, BSHYJDR high dosage group, low dosage BSHYJDR group, DDP combined with high and low dosage BSHYJDR group, 10 mice in each group. DDP group received injection intraperitoneally （ip） at the dosage of 1 mg/kg （equal to 1/10 LD50）, once a day for 4 d. High and low dosage BSHYJDR groups received intragastric BSHYJDR at the dosages of 26.6 and 13.3 g/kg （20 and 10 times each of clinical adult dosage） respectively, while tumor-bearing model group received the equal volume of distilled water once a day for 10 d. On the 11^th d, the mice were weighed and killed, then the tumor was dissected and weighed, the repression rate （RR） was calculated according to the mean weight of tumor （MWT）. RESULTS： Compared to the model group （MWT： 1.30±0.73）, DDP group （MWT： 0.41±0.09, RR： 68.46%） had a significant difference in the inhibition of hepatocarcinoma H22 （P〈0.01）. High dosage BSHYJDR group （MWT： 0.69±0.29, RR： 46.92%） also had a significant difference in inhibition （P〈0.05）, while no difference was found in low dosage BSHYJDR group （MVVT： 0.85±0.34, RR： 34.62%） （P〉0.05）. When DDP was combined with high dosage BSHYJDR （MWT： 0.29±0.17, RR： 77.69%） and low dosage BSHYJDR （MWT： 0.38±0.21, RR： 70.77%） respectively, we could see improvement of the inhibition effect of DDP on transplanted hepatocarcinoma H22. DDP combined with high dosage BSHYJDR had a significant difference （P〈0.001） compared to DDP, while DDP combined with low dosage BSHYJDR only had a little improvement that is not remarkable. CONCLUSION： Chinese medicine BSHYJDR in combination with chemotherapy can inhibit transplanted hepatocarcinorna in mice.

Intervention of Bushen Yizhi formula on cognitive disorder rat and the related mechanism of the protective function on neural network

OBJECTIVE Bushen Yizhi formula,constructed by Prof LIAO Shi-long′research group(Guangzhou University of chinese medicine),is the combination of clinical experience and modern pharmacological research,and mainly focuses on the etiology and pathogenesis of AD for treating both cause and symptoms.In this research,the pharmacodynamic study protocol of Bushen Yizhi formula was designed according to the characteristics and indications function of herbs in the formula,and was conformed to preclinical pharmacological research of traditional Chinese medicine for AD,The Technical Requirements of Pharmacology and Toxicology Research in New Drug Development Enacted by State Food and Drug administration also was guiding principle.METHODS A preliminary randomized double-blind clinical trail with 141 cases of AD showed that the efficiency of Bushen Yizhi formula was 61.9%and the increased mean of MMSE was 3.17,but the specific mechanism remains unclear.This study was aimed to reveal the preventive and therapeutic effect of Bushen Yizhi formula on AD rats and its related mechanism.In this research,forebrain-injected IBO-induced AD rats(cholinergic neuron lesion),senescence accelerated mouse,scopolamineinduced learning and memory deficiency rats(mild cognitive impairment)were all established.The learning and memory ability was tested with Morris water maze.Then formation of age pigment,extent of neuronal loss,activation of astrocytes,content of NTFs and degree of oxidized stress damage were determined by morphology,mmunohistochemical and molecular biology methods.RESULTS As the application of Bushen Yizhi formula,the learning and memory ability in all three groups were significantly improved,the formation of age pigment and the content of ACH in cortex and hippocampus were reduced,the activation of astrocytes and release of inflammatory factor(TNF-αand IL-1β)were inhibited,and the antioxidases(CAT,SOD,GSH-PX)were up-regulated and MDA was down-regulated.CONCLUSION Bushen Yizhi formula can prevent and treat AD rats,which might be achieved by anti-inflammatory,antioxidant and protecting cholinergic neuron.

Bushen Yizhi Formula regulates the IRE1αpathway to alleviate endoplasmic reticulum stress in an Alzheimer’s disease rat model

While the Bushen Yizhi Formula can treat Alzheimer’s disease(AD),the yet to be ascertained specific mechanism of action was explored in this work.Methods:Different concentrations of the Bushen Yizhi Formula and amyloid-beta peptide(Aβ)were used to treat rat pheochromocytoma cells(P12)and human neuroblastoma cells(SH-SY5Y).Cell morphological changes were observed to determine the in vitro cell damage.Cell Counting Kit(CCK)-8 assay and flow cytometry were employed to identify cell viability and apoptosis/cell cycle,respectively.Western blotting and immunohistochemistry were employed to measure the expressions of endoplasmic reticulum stress(ERS)-related proteins(GRP78 and CHOP),p-IRE1α,IRE1α,ASK1,p-JNK,JNK,Bax,Bcl-2,XBP-1,and Bim.Fura 2-acetoxymethyl ester(Fura-2/AM)was used to determine the intracellular calcium(Ca^(2+))concentration.Also,an AD model was constructed by injecting Aβinto the CA1 area of the hippocampus in Sprague Dawley rats.AD model rats were gavaged with different concentrations of Bushen Yizhi Formula for 14 consecutive days.The Morris water maze experiment was conducted to test the learning and memory of rats.Hematoxylin&Eosin(H&E)and Terminal-deoxynucleotidyl Transferase(TdT)-mediated dUTP Nick-End Labeling(TUNEL)staining were done to determine histopathological changes in the brain.Results:Bushen Yizhi Formula relieved the Aβ-induced effects including cell injury,decreased viability,increased apoptosis,G0/G1 phase cell cycle arrest,upregulation of GRP78,CHOP,p-IRE1α,p-JNK,Bax,XBP-1 and Bim,as well as down-regulation of Bcl-2.These results were also seen with IRE1αsilencing.While Aβsuppressed the learning and memory abilities of rats,the Bushen Yizhi Formula alleviated these effects of Aβ.Brain nerve cell injury induced by Aβcould also be treated with Bushen Yizhi Formula.Conclusion:Bushen Yizhi Formula could influence ERS through the IRE1αsignaling pathway to achieve its therapeutic effects on AD.

Mechanism of treating recurrent abortion with “Peiyuan Bushen Antai formula” based on network pharmacology and molecular docking

Objective:To study the effective compound action target signal pathway of Peiyuan bushenan taifang(PYBSATF)has achieved good clinical efficacy in the treatment of recurrent spontaneous abortion(RSA),but its mechanism has not been clarified because of its complex components.In this study,network pharmacology is applied to study the effective compounds,targets and signal pathways of PYBSATF in the treatment of RSA,so as to reveal its pharmacological mechanism of action in the treatment of recurrent spontaneous abortion.Methods:The Traditional Chinese Medicine Systems Pharmacology database(TCMSP)and CNKI are used to obtain the main compounds and drug action targets of PYBSATF.Genecards and the Online Mendelian Inheritance of Man databases(OMIM)are searched to collect the known genes related to RSA,so as to construct compound-target network and screen out the common target proteins and main active compounds.We also use string database to construct a visual protein-protein interaction network(PPI).Cluster Profiler and R software were used to analyze the common targets of drugs and diseases for GO function analysis and KEGG pathway enrichment analysis.Finally,the compound and the protein sequences were conducted according to the node parameters,so that the core protein and core compounds are used to perform molecular docking.Results:186 potential active components and 65 predicted action targets of PYBSATF were screened out.At the same time,1658 genes were also screened out to be closely related to RSA,among which 65genes overlaped with PYBSATF targets and were considered to be related to way of treatment.PPI network showed that VEGFA,IL6,EGFR,MAPK8 and ESR1were the core targets of PYBSATF for the treatment of RSA.GO and KEGG enrichment analysis obtained 93 biological processes of cells(P<0.01)and 87 signaling pathways(P<0.01).PYBSATF played a pharmacological role through a variety of pathways including anti-inflammatory,anti-apoptosis,promoting proliferation and angiogenesis.Molecular docking showed that most active components and key targets of PYBSATF had strong efficiency.Conclusion:Through the study of network pharmacology,it predicted that PYBSATF might treat RSA through multiple targets and multiple signal pathways.Significantly,the predictive targets may be potential targets for treatment of RSA.

Mechanism and Research Progress of Fuyang Huoxue Jiedu Formula against Recurrence of Ulcerative Colitis

By introducing the composition and modern pharmacology of Fuyang Huoxue Jiedu Formula,the paper further explored the mechanism and research progress of Fuyang Huoxue Jiedu Formula against recurrence of ulcerative colitis(UC).The formula can effectively improve the TCM syndromes of UC patients,reduce the recurrence rate,and improve clinical efficacy and patients’quality of life.

Effective ingredients, potential targets and mechanism in cancers treatment of Bushen Jiedu Sanjie recipe

Objective: To analyze the active compounds, potential drug targets and corresponding therapy cancer of Bushen Jiedu Sanjie Recipe (BSJDSJR) based on network pharmacology and bioinformatics technology. Methods: The network databases including Cancer HSP, TCMSP, TCMID, TCM-PTD, TCM Database@Taiwan and DrugBank were applied to screen the active compounds, potential drug targets and corresponding cancers of BSJDSJR. Cytoscape3.3 software was used to construct the network between active compounds of Chinese medicine and the corresponding targets. Then, the enrichment of biological processes and KEGG pathways of BSJDSJR were analyzed using DAVID database. Results: According to Oral bioavailability (OB)≥30% and drug like index (DL)≥0.18, 129 active compounds in BSJDSJR were screened out and they targeted to 301 proteins. These targets were closely associated with the occurrence of various cancers, such as bladder cancer, pancreatic cancer, non-small cell lung cancer and colorectal cancer. Further investigation showed that, there were lots of active compounds in BSJDSJR are closely connected with the COX-2/β-catenin signaling pathway, STAT3 signaling pathway and MAPK/ERK signaling pathway. Conclusions: Based on the network pharmacology, the study disclosed the active chemical compounds, potential targets and possible action cancers of BSJDSJR.

基于NKG2D及其配体探讨清热解毒方改善肝癌小鼠免疫微环境及抗肿瘤作用

目的:探讨清热解毒方对小鼠肝癌模型的抗肿瘤作用和淋巴细胞亚群、自然杀伤细胞家族2成员D(NKG2D)及其配体MULT1的影响。方法:建立小鼠H22肝癌模型,观察清热解毒方的抗肿瘤效应。采用流式细胞术检测小鼠外周血淋巴细胞亚群和肿瘤组织、瘤旁NKG2D及其配体MULT1表达情况的变化。结果:清热解毒方有一定抗肿瘤作用,且能改善荷瘤小鼠的体重降低;清热解毒方对小鼠外周血淋巴细胞亚群有一定影响,中药组较对照组B细胞、CD8^(+)T细胞比例降低,CD3^(+)T细胞比例、CD4^(+)T/CD8^(+)T增加(P<0.05);小鼠肝癌模型中,癌旁组织中NKG2D和MULT1阳性率均高于肿瘤组织,中药干预后,肿瘤组织和癌旁组织的NKG2D和MULT1阳性率均显著高于对照组(P<0.05)。结论:清热解毒方抗肿瘤机制可能与增强NKG2D及其配体介导的抗肿瘤免疫反应,调节T淋巴细胞亚群平衡,改善免疫微环境有关,而NKG2D及其配体系统可作为肿瘤治疗及预后评估的潜在靶点。

补肾活血方对抗磷脂抗体阳性反复种植失败患者妊娠结局的影响

目的:观察补肾活血方对抗磷脂抗体阳性反复种植失败(RIF)患者妊娠结局的影响。方法:选取188例抗磷脂抗体阳性RIF患者,按随机数字表法分为补肾活血方组和对照组,两组各94例。两组均给予冻融胚胎移植方案治疗,对照组同时口服拜阿司匹林治疗,治疗组在对照组基础上给予补肾活血方治疗。比较两组子宫内膜容受性超声检测指标、子宫内膜容受性分子标志物指标、中医证候积分及临床疗效、安全性。结果:两组基线时子宫内膜厚度及血流参数比较,无统计学差异(P>0.05)。两组胚胎移植前1天子宫内膜厚度及子宫内膜收缩期峰值速度/舒张末期流速(S/D)较基线时升高,子宫内膜搏动指数(PI)、阻力指数(RI)较基线时下降(P<0.05)。治疗组胚胎移植前1天子宫内膜厚度及子宫内膜S/D高于对照组,子宫内膜PI、RI低于对照组(P<0.05)。治疗组种植窗期整合素β3低于对照组,整合素β1、同源框基因11(HoxA11)高于对照组(P<0.05)。两组治疗后中医证候积分较治疗前降低,治疗组更低(P<0.05)。治疗组临床妊娠率62.22%、活产率55.56%高于对照组,胚胎种植率40.27%,流产率10.71%,与对照组比较,无统计学差异(P>0.05)。结论:补肾活血方可改善抗磷脂抗体阳性RIF患者子宫内膜容受性,提高临床妊娠率,降低早期流产率。

健脾补肾方对肌萎缩侧索硬化小鼠的干预效果及作用机制

目的探索健脾补肾方对肌萎缩侧索硬化(ALS)模型小鼠运动功能的影响及其机制。方法将12只采取转基因法繁育的保留部分运动功能的ADAR2flox/flox/VAChT-Cre小鼠(AR2小鼠)随机分为模型组和治疗组,每组6只;以6只同批次野生型(WT)小鼠为对照组。治疗组小鼠灌胃给予健脾补肾方[30 g/(kg·d)],对照组和模型组小鼠灌胃给予0.2 mL氯化钠溶液(9 g/L),每日1次,连续干预8周。每周观察小鼠活动状况、体质量以及抓力、抗疲劳能力(滚轮测试)等行为学功能。第24周后取样,采用苏木精-伊红(HE)染色法、尼氏染色法观察各组小鼠脊髓前角组织病理变化,采用Western blot法检测各组小鼠脊髓组织中作用于RNA的腺苷脱氨酶2(ADAR2)、转录反应DNA结合蛋白-43(TDP-43)的蛋白表达量,透射电镜观察小鼠脊髓组织超微结构。结果与对照组比较,模型组小鼠体质量随周龄逐渐增长;与模型组比较,治疗组小鼠体质量降低。与对照组比较,模型组小鼠与治疗组小鼠抗疲劳程度均有所下降,但是治疗组小鼠的抗疲劳能力下降相对模型组有所延缓。与对照组比较,模型组小鼠抓力下降;与模型组比较,治疗组抓力下降相对减缓。与对照组相比,模型组TDP-43表达量显著降低(P<0.001);与模型组相比,治疗组TDP-43表达量显著升高(P<0.05)。与对照组相比,模型组ADAR2表达量显著降低(P<0.01),治疗组与模型组之间ADAR2表达量没有显著差异(P>0.05)。与对照组相比,模型组脊髓前角运动神经元损伤严重,神经元结构不完整;与模型组相比,治疗组神经细胞凋亡数量减少。透射电镜下,模型组与对照组比较出现明显的线粒体肿胀凋亡、髓鞘脱散现象;治疗组与对照组相比较存在部分髓鞘脱散、线粒体肿胀,相对于模型组,存在肿胀凋亡现象的神经细胞数量有所减少。结论健脾补肾方能延缓ALS模型小鼠运动功能的减退,减少脊髓运动神经元的凋亡和损伤,阻止散发性ALS(sALS)病理异常标志物TDP-43的生成,保护脊髓神经元有髓神经纤维与线粒体,有效延缓ALS小鼠疾病的进展与功能的丧失。

基于METTL14-m6A-FOXO3A信号通路介导的细胞自噬探讨补肾强督方抑制炎症治疗强直性脊柱炎的机制研究

目的:观察补肾强督方对T淋巴细胞METTL14-m^(6)A-FOXO3A信号通路介导的细胞自噬的影响,从而明确其抑制炎症、治疗强制性脊柱炎的分子机制。方法:使用Jurkat细胞,采用抗CD3联合抗CD28抗体诱导24 h构建细胞模型,并给予补肾强督方含药血清和METTL14重组蛋白进行干预。采用ELISA法检测炎症因子(IL-23和IL-17A)和氧化应激指标的水平,流式细胞术检测ROS水平,Western blot法检测自噬相关蛋白、METTL14和FOXO3A的表达,RT-qPCR检测METTL14和FOXO3A的表达,斑点杂交实验检测FOXO3A的m^(6)A甲基化水平。结果:与模型组比较,补肾强督方显著降低Jurkat细胞上清中IL-23、IL-17A和MDA的含量与ROS的水平,增加SOD和GSH-Px的活性,差异均具有统计学意义(P<0.01)。此外,补肾强督方能显著上调Jurkat细胞中Beclin-1、FOXO3A和METTL14蛋白和基因的表达和增加LC3Ⅱ/LC3Ⅰ的比值,下调p62蛋白的表达,与模型组比较差异均具有统计学意义(P<0.01)。在此基础之上,与模型组比较,补肾强督方能显著上调Jurkat细胞FOXO3A的m^(6)A甲基化水平,差异均具有统计学意义(P<0.01)。结论:补肾强督方通过METTL14介导的FOXO3A的m^(6)A甲基化修饰促进T细胞自噬,从而达到治疗强直性脊柱炎的作用。

补肾调经方联合脱氢表雄酮治疗卵巢储备功能减退临床研究

目的:探究自拟补肾调经方联合脱氢表雄酮(DHEA)治疗卵巢储备功能减退(DOR)的临床疗效。方法:选择74例观察期未进行激素补充治疗(HRT)的DOR患者,其中试验组40例予以补肾调经方联合DHEA治疗;对照组34例予以DHEA治疗,疗程均为3个月经周期,比较两组患者的性激素(FSH、LH、E2、T)、抗缪勒管激素(AMH)、卵巢体积、基础窦卵泡(AFC)、改良Kupperman量表总积分及中医症状评分的变化。结果:试验组治疗后FSH明显低于治疗前及同期对照组(P<0.05),而LH、E2、T及AMH在两组间差异无统计学意义(P>0.05);试验组治疗后AFC明显高于治疗前及同期对照组(P<0.05),而卵巢体积两组间差异无统计学意义(P>0.05);试验组治疗后改良Kupperman量表总积分明显低于治疗前及同期对照组(P<0.05);试验组治疗后部分中医症状评分明显低于治疗前及同期对照组(P<0.05)。结论:补肾调经方联合DHEA治疗DOR可获得满意疗效,明显优于单独DHEA治疗。

邓悦教授应用自拟豁痰解毒通络方治疗冠心病合并颈动脉狭窄1例

冠心病合并颈动脉狭窄是一种常见的心血管疾病。西医的治疗取得了一定的进展。邓悦教授应用自拟豁痰解毒通络方治疗胸痹心痛脉痹合病,并提出气虚、痰浊、瘀血是此病的根本病机,“痰瘀伏邪,热结血络”是其重要的病机变化,与“膏粱之变,足生大丁”的病机有异曲同工之处。采用益气化瘀、豁痰解毒的治法,在临床中取得了显著的治疗效果。因此,总结了邓悦教授在治疗冠心病合并颈动脉狭窄方面的用药经验,为临床提供一定的参考。

余仁欢辨治药物性肾损害经验

药物性肾损害作为临床用药较为常见的不良反应事件,在治疗上具有一定难度。余仁欢教授认为治疗药物性肾损害应重点关注损伤程度、损伤部位、急慢性质以及患者素体情况等。对于平素体健猝逢药毒攻伐的实证患者,以解毒泻浊化瘀法治之;对于脾肾虚甚、不耐药之毒性或偏性的虚证患者,以补脾益肾法治之;对于脾肾素亏又逢药毒攻伐致脾肾两虚、痰凝湿阻、络损血瘀的虚实夹杂证患者,以健脾祛湿、化瘀和络法治之。附验案1则。

醒脑开窍针刺法合并补肾化痰活血方治疗急性脑梗死风痰瘀阻证的临床研究

目的:探究醒脑开窍针刺法合并补肾化痰活血方治疗急性脑梗死风痰瘀阻证的临床效果。方法:选择北京中医药大学孙思邈医院2021年1月-2023年12月收治的急性脑梗死风痰瘀阻证患者106例,采用随机数字表法分为针刺组和联合组,每组53例。两组均给予抗凝、改善微循环、控制血压、神经保护、吸氧、心脏监测等常规西医治疗,针刺组给予醒脑开窍针刺法治疗,联合组在针刺组基础上给予补肾化痰活血方口服治疗。观察两组患者的中医证候积分、神经功能指标、血流动力学指标、神经功能与运动功能及日常生活能力评分、临床疗效、不良反应发生情况。结果:治疗后,两组中医证候积分、美国国立卫生研究院卒中量表(NIHSS)评分、神经胶质纤维酸性蛋白(GFAP)、神经元特异性烯醇化酶(NSE)、中枢神经特异蛋白(S100β)、血流峰值时间(TTP)与阻力指数(RI)水平均较治疗前降低(P<0.05),且联合组低于针刺组(P<0.01);治疗后,两组血清神经生长因子(NGF)、脑血容量(CBV)、平均血流量(Qmean)水平、简式Fugl-Meyer评分(FMA)与Barthel指数(BI)评分均较治疗前升高(P<0.05),且联合组高于针刺组(P<0.01);联合组治疗总有效率高于针刺组(P<0.05);两组不良反应总发生率差异无统计学意义(P>0.05)。结论:醒脑开窍针刺法合并补肾化痰活血方治疗急性脑梗死风痰瘀阻证可降低中医证候积分,缓解临床症状,增加脑血流量,改善神经功能,提高临床疗效,且安全性较高。

补肾活血方联合西药治疗肾虚血瘀型排卵障碍性不孕患者的效果

目的观察补肾活血方联合枸橼酸氯米芬治疗肾虚血瘀型排卵障碍性不孕患者的效果。方法选取2020年1月至2023年5月延安市中医医院收治的肾虚血瘀型排卵障碍性不孕患者80例进行随机对照试验。采用随机数字表法将其分为试验组和对照组,各40例。对照组年龄(26.87±9.55)岁,病程(1.95±0.63)年,体重指数(24.87±3.58)kg/m2,基础体温(36.12±1.32)℃,月经量(17.12±2.67)ml。试验组年龄(27.62±8.79)岁,病程(2.04±0.50)年,体重指数(25.62±2.59)kg/m2,基础体温(36.25±0.97)℃,月经量(16.27±3.85)ml。对照组给予枸橼酸氯米芬治疗,试验组在此基础上加用补肾活血方。每个疗程5 d,两组均治疗8个疗程。记录两组治疗前后子宫内膜厚度、卵泡数、卵泡长径、血清促卵泡激素(follicle-stimulating hormone,FSH)、黄体生成素(luteinizing hormone,LH)和雌二醇(estradiol,E2)水平、疗效及不良反应发生情况。采用t检验和χ^(2)检验进行统计学分析。结果治疗后,试验组子宫内膜厚度、卵泡数、卵泡长径均高于对照组[(9.17±3.62)mm比(7.07±2.06)mm、(6.80±2.57)个比(4.19±2.66)个、(16.81±3.42)mm比(15.29±3.19)mm;t=3.189、4.463、2.056,均P<0.05]。治疗后,试验组血清FSH、LH、E2水平均高于对照组[(9.91±2.56)IU/L比(8.82±1.69)IU/L、(10.61±2.39)IU/L比(7.62±3.31)IU/L、(89.12±13.46)ng/L比(65.14±15.56)ng/L;t=2.247、4.623、7.372,均P<0.05]。试验组治疗总有效率高于对照组[85.00%(34/40)比57.50%(23/40);χ^(2)=6.102,P=0.014]。试验组总不良反应发生率低于对照组[5.00%(2/40)比27.50%(11/40);χ^(2)=5.878,P=0.015]。结论补肾活血方联合枸橼酸氯米芬治疗肾虚血瘀型排卵障碍性不孕患者的效果较好。

自拟解毒疏肝方治疗原发性肝癌的临床效果研究

目的观察自拟解毒疏肝方治疗原发性肝癌的临床效果。方法60例肝癌患者,随机分为观察组和对照组,各30例。对照组患者采用抗炎保肝治疗,观察组在上述治疗的基础上,口服中汤药自拟解毒疏肝方。比较两组患者的生存率及生活质量、中医证候积分、甲胎蛋白水平。结果两组治疗3个月生存率对比,差异无统计学意义(P>0.05),观察组治疗6、12个月生存率分别为90.0%、73.3%,均高于对照组的66.7%、46.7%,差异具有统计学意义(P<0.05)。治疗前、治疗3个月,两组患者生活质量评分对比,差异无统计学意义(P>0.05);治疗6、12个月,观察组患者生活质量评分分别为(65.7±6.3)、(68.3±5.9)分,均高于对照组的(57.7±9.0)、(56.3±8.5)分,差异有统计学意义(P<0.05)。治疗6、12个月,观察组患者中医证候积分分别为(53.9±1.3)、(50.5±6.7)分,均低于对照组的(61.1±4.4)、(66.4±8.3)分,差异有统计学意义(P<0.05)。治疗6、12个月,观察组患者甲胎蛋白水平分别为(354.0±24.0)、(332.0±9.4)ng/ml,低于对照组的(422.3±7.5)、(446.5±16.1)ng/ml,差异有统计学意义(P<0.05)。结论自拟解毒疏肝方结合西医治疗肝癌可提高临床疗效,改善患者生活质量,同时降低甲胎蛋白水平,安全可靠,值得临床应用推广。

清热解毒方联合纤维支气管镜治疗儿童难治性肺炎支原体肺炎痰热闭肺证的效果及对肺功能、炎性因子和免疫功能的影响

目的探究清热解毒方联合纤维支气管镜治疗难治性肺炎支原体肺炎(RMPP)痰热闭肺证患儿的效果及对肺功能、炎性因子和免疫功能的影响。方法选取2024年1月至2024年8月收治的106例RMPP痰热闭肺证患儿,依据不同治疗方案分为观察组(清热解毒方联合纤维支气管镜治疗)和对照组(纤维支气管镜治疗)各53例。2组治疗2周后评估治疗效果和支气管黏膜损伤修复情况,比较2组治疗前、治疗2周后肺功能指标、中医证候积分、炎性因子、免疫功能指标以及不良反应发生情况。结果治疗2周后,观察组愈显率高于对照组(P<0.05),且观察组治疗后支气管黏膜修复效果优于对照组。治疗2周后,2组最大呼气流量、第一秒用力呼气容积、用力肺活量较治疗前升高,且观察组高于对照组(P<0.05)。2组治疗2周后咳嗽、发热、咳黄黏痰、气喘积分低于治疗前,且观察组各项积分均低于对照组(P<0.05,P<0.01)。治疗2周后,2组血清肿瘤坏死因子-α、淀粉样蛋白A/C反应蛋白、白细胞介素-23、γ-干扰素水平较治疗前降低,且观察组较对照组低(P<0.05,P<0.01)。2组治疗2周后CD3+、免疫球蛋白A、CD4+/CD8+、免疫球蛋白G水平高于治疗前,且观察组高于对照组(P<0.05,P<0.01)。2组治疗期间不良反应总发生率比较差异无统计学意义(P>0.05)。结论清热解毒方联合纤维支气管镜治疗能抑制RMPP痰热闭肺证患儿体内炎症反应,改善临床症状及免疫功能,增强肺功能,且安全较好。

Expression of Nrf2/ARE pathway in diabetic nephropathy and renal protection of Qihuang Bushen Xiezhuo Formula

Objective:To investigate the effect of Qihuang Bushen Xiezhuo Formula on the expression of the nuclear factor E2 related factor 2(Nrf2)/antioxidant response element(ARE)signaling pathway of human glomerular mesangial cells(HMC)in high glucose medium and its protective effect on oxidative stress.Methods:The HMC were cultured in vitro to prepare normal rat serum.The rats were intragastrically administered with irbesartan and Qihuang Bushen Xiezhuo Formula.The serum containing the drugs was prepared after the blood concentration was reached.The rats were divided into the control group,the high glucose group,the irbesartan group and the Qihuang Bushen Xiezhuo Formula group.The HMC of the control group was cultured with normal rat serum(10%serum concentration)medium,while that of the high glucose group was cultured with high glucose medium of rat serum(10%serum concentration).The irbesartan group and the Qihuang Bushen Xiezhuo Formula group were respectively treated with 10%irbesartan and 10%Qihuang Bushen Xiezhuo Formula in serum high glucose medium.After 48 h of culture,the relevant indicators were collected and detected.The mRNA expression levels of Nrf2,gamma-glutamylcysteine synthetase(γ-GCS)and superoxide dismutase(SOD)in each group were detected by real-time PCR.The expressions ofγ-GCS and SOD were detected by immunohistochemistry.The protein expressions ofγ-GCS and SOD in HMC of each group were observed by Western Blot.Results:The expressions of Nrf2,γ-GCS,SOD mRNA and protein in experimental cells of each group were low in the control group,while those in the high glucose group were decreased as compared with the control group(P<0.05).After interference of irbesartan and Qihuang Bushen Xiezhuo Formula the expressions were increased,and the increase in Qihuang Bushen Xiezhuo Formula group was more significant(P<0.05).Conclusion:Qihuang Bushen Xiezhuo Formula can improve HMC oxidative stress injury in high glucose culture,and its mechanism may be achieved by activating Nrf2 and its related downstream proteinsγ-GCS and SOD.

基于JNK通路探讨补肾活血方对大鼠骨关节炎软骨细胞凋亡的影响

目的 探讨补肾活血方(Bushen Huoxue Formula,BSHXF)通过c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)信号通路对大鼠骨关节炎(Osteoarthritis,OA)软骨细胞凋亡的影响。方法 采用白细胞介素-1β (interleukin-1β,IL-1β)诱导体外分离培养的大鼠软骨细胞构建体外OA模型,并使用不同浓度的BSHXF干预处理。使用MTT法测定细胞活力;CCK-8法检测细胞增殖能力;流式细胞术评估细胞凋亡情况;Western bolt检测凋亡相关蛋白裂解的胱天蛋白酶-3(Cleaved Caspase-3)、B淋巴细胞瘤-2(B-cell lymphoma 2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2-associated X protein,Bax)水平和JNK、磷酸化c-Jun氨基末端激酶(phosphorylated c-Jun N-terminal kinase,p-JNK)蛋白水平。试剂盒检测细胞中活性氧(reactive oxygen species,ROS)、丙二醛(malondialdehyde,MDA)、谷胱甘肽(Glutathione,GSH)水平;ELISA法检测细胞上清液中促炎因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)含量。结果 (1)与对照组相比,模型组软骨细胞活力显著降低,细胞凋亡率显著升高(P<0.01);模型组Cleaved Caspase-3、Bax、p-JNK蛋白表达水平显著升高(P<0.01),Bcl-2蛋白表达水平显著降低(P<0.01)。与模型组相比,BSHXF治疗组软骨细胞活力升高(P<0.05),凋亡率显著降低(P<0.01);BSHXF治疗组Cleaved Caspase-3、Bax、p-JNK蛋白表达水平显著降低(P<0.01),Bcl-2蛋白表达水平升高(P<0.01)。(2)与对照组相比,模型组细胞中ROS和MDA的相对含量显著升高(P<0.01),GSH的相对含量显著降低(P<0.01);模型组细胞中TNF-α和IL-6的含量均显著升高(P<0.01)。与模型组相比,BSHXF治疗组细胞中ROS和MDA的相对含量显著降低(P<0.01),GSH的相对含量显著升高(P<0.01);BSHXF治疗组细胞中TNF-α和IL-6的含量均显著降低(P<0.01)。(3)与溶剂对照组相比,激活剂对照组的p-JNK、Cleaved Caspase-3、Bax蛋白水平显著上调(P<0.05),Bcl-2蛋白表达水平显著降低(P<0.01),细胞增殖能力下降(P<0.05),细胞凋亡率显著增加(P<0.01);BSHXF治疗组和溶剂对照组两组间细胞增殖、凋亡率和凋亡相关蛋白表达(Cleaved Caspase-3、Bax、Bcl-2)情况差异均无统计学意义(P>0.05)。(4)与溶剂对照组相比,激活剂对照组的ROS和MDA水平显著升高(P<0.01),而GSH水平显著降低(P<0.01),TNF-α、IL-6含量升高(P<0.05);BSHXF治疗组和溶剂对照组两组间ROS相对含量、MDA、GSH水平差异均无统计学意义(P>0.05)。结论 BSHXF通过抑制JNK信号通路激活抑制IL-1β诱导的OA软骨细胞凋亡,改善氧化应激和炎症反应。

补肾活血方调控NF-κB信号通路对膝骨关节炎小鼠软骨组织炎症及细胞凋亡的影响

目的观察补肾活血方(BSHXF)对膝骨关节炎(KOA)小鼠软骨组织炎症及细胞凋亡的影响,探讨BSHXF治疗KOA的作用机制。方法将24只雄性BALB/c小鼠按体重随机分为假手术组、KOA模型组、KOA模型+BSHXF组,每组8只。造模、灌胃干预后,小鼠取血,ELISA检测血清IL-6、iNOS、COX2水平;提取小鼠膝关节软骨组织,HE染色及番红O-固绿染色观察软骨组织病理学情况,TUNEL染色检测软骨组织细胞凋亡情况,Western blot检测Cleaved-Caspase-9、Bcl-2、Bax蛋白表达,qRT-PCR检测CollagenⅡ、Aggrecan、ADAMTS-5 mRNA表达,Western blot检测核转录因子-κB(NF-κB)信号通路关键蛋白p-IκBα、IκBα、p-p65、p65的表达。结果与假手术组相比,KOA模型组小鼠膝关节软骨细胞病理改变明显;血清炎症细胞因子IL-6、iNOS和COX2的水平升高,软骨组织细胞凋亡率增加,Cleaved-Caspase-9、Bax蛋白表达水平升高,Bcl-2蛋白表达降低,CollagenⅡ、Aggrecan的mRNA表达水平降低,ADAMTS-5 mRNA表达升高,p-IκBα/IκBα、p-p65/p65比值升高。与KOA模型组相比,KOA模型+BSHXF组膝关节软骨损伤改善、病理变化减轻,炎症细胞因子IL-6、iNOS和COX2的水平降低,软骨组织细胞凋亡率降低,Cleaved-Caspase-9、Bax蛋白表达降低,Bcl-2蛋白表达升高,CollagenⅡ、Aggrecan的mRNA表达升高,ADAMTS-5 mRNA表达降低,p-IκBα/IκBα、p-p65/p65比值降低。结论BSHXF可抑制NF-κB信号通路,减少炎性因子释放、抑制软骨细胞凋亡及细胞外基质降解,从而缓解KOA疾病进展。