Meta-analysis of the curative efficacy of Duhuo Jisheng Decoction combined with western medicine on knee osteoarthritis

Objective:To evaluate the efficacy and safety of Duhuo Jisheng Decoction combined with western medicine in the treatment of knee osteoarthritis.Methods:The databases of CNKI,VIP,CBM,WANFANG DATA and PubMed were electronically to collect randomized controlled trials(RCTs)of the treatment of knee osteoarthritis with Duhuo Jisheng Decoction combined with western medicine.After literature was strictly screened and the study quality was evaluated according to relevant procedures,the Meta analysis was performed by RevMan5.3 software.Resluts:A total of 15 RCTs were included.The result of meta-analysis showed that:compared with the western medicine alone,Duhuo Jisheng Tang combined with western medicine could significantly improve the efficacy(RR=1.25,95%CI(1.18,1.32),p<0.00001),the posttreatment pain visual analogue scale scores(MD=-0.85,95%CI(-1.00,-0.71),p<0.00001)、the western ontario and mcmaster universities arthritis index scores(MD=-7.76,95%CI(-10.37,-5.15),p<0.00001)and matrix metalloproteinase3(MD=-11.72,95%CI(-17.39,-6.05),p<0.0001)were lower in treatment group compared to control group.There was no statistical difference in the incidence of adverse reactions between the 2groups.Conclusions:The current evidence shows that Duhuo Jisheng Decoction combined with western medicine can further improve the clinical efficacy for knee osteoarthritis.

Meta-analysis of clinical efficacy of Duhuo Jisheng Decoction in the treatment of lumbar intervertebral disc herniation

Objective:To explore the clinical effect of Duhuo Jisheng Decoction in the treatment of lumbar disc herniation.Methods:CNKI,VIP,Wan Fang Data,Pubmed,Cochrane Library,Medline Complete and Embase databases were searched by computer.The randomized controlled trial(RCT)of Duhuo Jisheng Decoction combined with western medicine in the treatment of lumbar intervertebral disc herniation was studied.The searching time was from the establishment of the database to July 20,2019.Data were extracted and analyzed by Revman 5.3 software.Results:Twenty-four literatures involving 1980 patients were included.Meta-analysis showed that the total effective rate of Duhuo Jisheng Decoction group was significantly higher(OR=5.26,95%CI(4.13,6.69),Z=13.47,P<0.00001)and the incidence of adverse reactions was significantly lower(OR=0.45,95%CI(0.23,0.87),Z=2.36,P=0.00001)than that of Western medicine group.02).Conclusion:Duhuo Jisheng Decoction is effective and safe in the treatment of lumbar intervertebral disc herniation.However,the literature included in this study is limited,and the quality of the literature is low.It is necessary to carry out in-depth verification of high-quality,large-sample RCT.

A Systematic Review and Meta-analysis for Duhuo Jisheng Decoction Combined with Manipulation in Treatment of Lumbar Disc Herniation

Objective:To systematically evaluate the efficacy and safety of Duhuo Jisheng decoction combined with manipulation in the treatment of lumbar disc herniation.Method:We systematically searched 8 databases at home and abroad.Literature quality of the included studies was evaluated by the bias risk assessment tool of Cochrane collaboration network,and the Meta-analysis was performed by RevMan 5.3 software.Result:A total of 427 related literature were retrieved,and 12 studies were eventually included,involving 1153 patients.All studies were in Chinese.Meta-analysis results showed that the total effective rate of Duhuo Jisheng decoction combined with manipulation in the treatment of LDH was better than that of the control group(RR=1.23,95%CI[1.17,1.29],P<0.00001).Subgroup analysis showed that Duhuo Jisheng decoction combined with manipulation had a more significant effect than conventional treatment(RR=1.34,95%CI[1.20,1.50],P<0.00001),nonsteroidal anti-inflammatory drugs(RR=1.24,95%CI[1.12,1.37],P<0.0001),and Yaotongning capsules(RR=1.19,95%CI[1.10,1.29],P<0.0001).It has obvious advantages in improving VAS scores(MD=-1.19,95%CI-1.19[-1.87,-0.51],P=0.0006),but it has no obvious advantages in improving JOA scores(MD=2.6595%CI[0.06,5.24],P=0.05).In terms of safety,Duhuo Jisheng decoction combined with manipulation treatment reported fewer adverse events.Conclusion:The Duhuo Jisheng decoction combined with manipulation treatment can effectively relieve the clinical symptoms of patients with lumbar disc herniation,with obvious clinical efficacy,and fewer adverse events.But in view of the limitations of clinical studies,there are some limitations in this study,and more studies with high quality of evidence are needed to further confirm its efficacy and safety.

A meta-analysis of Duhuo Jisheng decoction combined with non-operative therapy on lumbar disc herniation

Objective:To systematically evaluate the cllinical efficacy of Duhuo Jisheng decoction(DHJSD)combined with non-surgical therapy on lumbar disc herniation(LDH).Methods:China Science and Technology Journal Database(VIP),Chinese National Knowledge Infrastructure(CNKI),WanFang Data,PubMed were searched and eligible randomized controlled clinical trials(RCTs)were included.Two reviewers evaluated the quality of the included RCTs and extracted data independently.Then,data analysis was performed with RevMetn 4.2 software.Results:A total of 18 RCTs were enrolled,including 1682 patients.Meta-analysis results showed that the overall response rate of DHJSD combined with non-operative therapy group(treatment group)in the treatment of LDH was better than that of the non-operative therapy group(control group)[OR=4.29,95%CI(3.19,5.76),P<0.001].JOA score of the treatment group was significantly higher than that of control group[OR=3.96,95%CI(3.4,4.52),P<0.001].VAS score was dramatically lower in the treatment group than the control group[OR=-1.58,95%CI(-1.97,-1.18),P<0.001].Conclusion:The clinical efficacy of DHJSD combined with non-surgical therapy is better than that of non-surgical therapy alone.However,due to the low quality of the included literatures,large-scale and high-quality RCTs are still needed for further confirmation.

Mechanism of Duhuo Jisheng Decoction based on bioinformatics and network pharmacology in treating osteoporosis of kidney Yin deficiency

Objective:To study the active ingredients,target and mechanism of Duhuo JiSheng Decoction for the treatment of osteoporosis by bioinformatics analysis and network pharmacology.Methods:The gene expression profile of GSE56116 was acquired through the NCBI GEO database,and the software R was used for differential analysis.The Chinese medicine system pharmacology technology platform(TCMSP)was used to retrieve the chemical components,target genes of Duhuo,Fangfeng,Xixin,and Sangjisheng.The differential genes acquired from GSE56116 were intersected and analyzed by GO enrichment analysis and KEGG pathway to study the mechanism of Duhuo JiSheng Decoction.Results:A total of 33 active ingredients and 9 differential genes of Duhuo JiSheng Decoction for the treatment of Shengyin deficiency osteoporosis were screened.GO function enrichment analysis focuses on the regulation of intracellular signal transduction,calcium-mediated signaling,and steroid hormone mediated signaling.KEGG analysis showed that it was mainly concentrated in signal pathways such as P53 signal pathway,TNF signal pathway,and IL-17 signal pathway.Conclusion:Through bioinformatics and network pharmacological analysis,Duhuo JiShenng Decoction may exert therapeutic effects by acting on AR,IGFBP3,and FOS target genes.

从“络病-风药”理论探讨独活寄生汤治疗类风湿关节炎

类风湿关节炎(RA)属中医“痹病”范畴,其发生、发展、转归与络病密切相关。本文通过分析类风湿关节炎微小血管新生的病理状态与络脉生理和络病病机特点,以络脉为其邪伏、邪发、邪伤及邪阻之所展开论述,可认为“络病”是其病理基础,络虚是其根本病机,风挟寒湿之邪内侵络脉为发病关键。基于“络病”理论和“风药”理论相关认识,从独活寄生汤为治风之剂视角认识其治疗类风湿关节炎的作用,并提出病变早期络脉郁滞、风药先行,中期络脉瘀塞、健脾化瘀,晚期络脉空虚、风引入肾,探讨独活寄生汤在病变不同时期风药与他药的组方原理,丰富独活寄生汤的组方配伍理论,扩展独活寄生汤的临床指导意义,为其临床应用提供参考。

独活寄生汤对TNF-α诱导的类风湿性关节炎成纤维样滑膜细胞增殖、凋亡和炎症的影响

目的探究独活寄生汤含药血清对肿瘤坏死因子-α(TNF-α)诱导的类风湿性关节炎(RA)成纤维样滑膜细胞(FLS)增殖、凋亡和炎症的影响及作用机制。方法将MH7A细胞分为正常组(10%空白血清)、模型组(10μg·L^(-1) TNF-α+10%空白血清)、独活寄生汤低(10μg·L^(-1) TNF-α+2.5%含药血清+7.5%空白血清)、中(10μg·L^(-1) TNF-α+5%含药血清+5%空白血清)、高(10μg·L^(-1) TNF-α+10%含药血清)剂量组共5组;MTT法筛选独活寄生汤含药血清给药浓度;EdU染色检测细胞增殖情况;免疫荧光染色检测增殖标志物Ki67表达情况;流式细胞仪检测细胞凋亡率;ELISA法检测细胞中IFN-γ、IL-4、IL-6和IL-10的含量;RT-qPCR和Western blot检测Bax、Bcl-2、caspase-3及TLR4/MyD88/NF-κB信号通路mRNA和蛋白的表达情况。结果与正常组比较,模型组细胞增殖水平明显上升,凋亡水平明显减弱,IFN-γ和IL-6的含量升高,IL-4和IL-10的含量减少,TLR4/MyD88/NF-κB信号通路被激活;经独活寄生汤含药血清低、中、高剂量组干预后,均能有效改善细胞异常增殖情况,增强细胞凋亡,抑制炎性反应和TLR4/MyD88/NF-κB信号通路的激活。结论独活寄生汤含药血清能够抑制TNF-α诱导的RA-FLS异常增殖和促炎因子的分泌,增强细胞凋亡和抗炎水平,其作用机制可能与调控TLR4/MyD88/NF-κB信号通路有关。

独活寄生汤含药血清对膝关节OA大鼠滑膜细胞TNF-α、IL-6、IL-8、IL-10表达的影响及其机制

目的探讨独活寄生汤含药血清对膝关节骨关节炎(OA)大鼠滑膜细胞炎症因子表达的影响及其可能机制。方法采用Hulth法建立OA大鼠模型,分离膝关节滑膜细胞,采用酶原消化法进行培养,并将其分为模型组及低、中、高剂量组,以正常大鼠膝关节滑膜细胞为对照组。低、中、高剂量组分别加入经独活寄生汤中药0.85、1.7、3.4 m L/kg灌胃大鼠的含药血清,对照组与模型组加入等量20%FBS,培养48 h。采用ELISA法检测各组细胞TNF-α、IL-6、IL-8及IL-10表达,超高效液相色谱-高分辨质谱法检测各组细胞脂氧素A4表达,Western blotting法检测各组细胞抗c-Jun氨基末端激酶(JNK)、磷酸化JNK(p-JNK)、激活子蛋白1(AP-1)及磷酸化AP-1(p-AP-1)表达。结果对照组、高剂量组、中剂量组、低剂量组和模型组细胞TNF-α、IL-6、IL-8表达均依次升高,IL-10、脂氧素A4表达依次降低,组间两两比较P均<0.05。与对照组比较,模型组细胞AP-1、p-AP-1、JNK及p-JNK表达均升高(P均<0.05);与模型组比较,高、中、低剂量组细胞AP-1、p-AP-1、JNK及p-JNK表达均降低,且高剂量组降低更明显(P均<0.05)。结论独活寄生汤含药血清可降低膝关节OA大鼠滑膜细胞TNA-α、IL-6、IL-8、IL-10表达,并呈剂量依赖性;提高滑膜细胞脂氧素A4表达、抑制JNK、AP-1及其磷酸化通路可能是其作用机制。

独活寄生汤含药血清对兔退变软骨细胞“caveolin-p38MAPK”信号通路调控作用的影响

目的:观察独活寄生汤含药血清对兔退变软骨细胞"caveolin-p38MAPK"信号通路的调控作用,探讨独活寄生汤治疗骨关节炎的作用机制。方法:将20只3月龄新西兰兔随机分为生理盐水组(空白血清组)和独活寄生汤组(含药血清组),每组10只。分别在24 h、36 h、48 h不同采血时间点采集独活寄生汤含药血清和空白血清,将5%、10%、15%、20%不同浓度两种血清作用于体外培养第2代软骨细胞,确定含药血清最佳干预条件;建立体外退变软骨细胞模型,分别给予独活寄生汤含药血清(含药血清组)和空白血清(空白血清组)干预36 h,收集软骨细胞,运用Western Blot法检测血清干预后退变软骨细胞caveolin-1、p38、p-p38蛋白表达,RT-PCR法检测血清干预后退变软骨细胞白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α、基质金属蛋白酶(MMP)-3、MMP-13、caveolin-1 m RNA表达。结果:浓度为15%的36 h采血时间点含药血清的促增殖作用最明显;退变软骨细胞中存在"caveolin-p38MAPK"信号通路的激活,独活寄生汤含药血清可抑制caveolin-1、p-p38蛋白表达及IL-1β、TNF-α、MMP-3、MMP-13、caveolin-1 m RNA的表达,差异有统计学意义(P<0.05)。结论:独活寄生汤能通过抑制"caveolin-p38MAPK"信号通路的激活及其下游效应分子,从而有效抑制软骨细胞凋亡。

独活寄生汤含药血清对大鼠退变软骨细胞Bcl-2、Bax表达的影响

目的:探讨独活寄生汤含药血清对大鼠退变软骨细胞Bcl-2、Bax表达的影响。方法:建立体外退变软骨细胞模型,分别给予独活寄生汤含药血清(含药血清组)和空白血清(空白血清组)干预24 h、48 h、72 h,收集软骨细胞,运用RT-PCR法检测血清干预后退变软骨细胞Bcl-2、Bax m RNA表达,Western Blot法检测血清干预后退变软骨细胞Bcl-2、Bax蛋白表达。结果:独活寄生汤含药血清对Bcl-2和Bax蛋白表达的影响与其对Bcl-2和Bax m RNA的影响具有相同的趋势,随着干预时间的延长,空白血清组和含药血清组Bcl-2蛋白阳性表达率均逐渐升高,含药血清组升高更显著;Bax蛋白阳性表达率逐渐降低,含药血清组降低更明显,且各时点之间比较,差异有统计学意义(P<0.01)。结论:独活寄生汤能通过上调大鼠骨关节炎软骨细胞抑制凋亡因子Bcl-2表达,下调促凋亡因子Bax表达,抑制线粒体凋亡通路的激活,从而有效抑制软骨细胞凋亡。

独活寄生汤对膝骨关节炎模型大鼠NF-κB通路关键分子表达的影响

目的:探讨独活寄生汤对膝骨关节炎模型大鼠核因子κB(nuclear factor-kappaB,NF-κB)通路关键分子表达的影响。方法:将60只SPF级Wistar大鼠分为空白组(10只)和造模组(50只)。模型制备成功后按随机数字表法分为模型组,硫酸氨基葡萄糖组(阳性组),独活寄生汤高、中、低剂量组,共5组,每组10只。空白组及模型组大鼠给予生理盐水灌胃,阳性组及独活寄生汤各剂量组给予等体积药物灌胃,连续灌胃21天。观察各组大鼠一般生存状况,苏木精-伊红染色法观察各组大鼠软骨组织病理变化,酶联免疫吸附法检测大鼠血清白细胞介素4(interleukin-4,IL-4)、白细胞介素6(interleukin-6,IL-6)、白细胞介素10(interleukin-10,IL-10)及肿瘤坏死因子α(tumor necrosis factorα,TNF-α)的含量,荧光定量PCR(quantitative polymerase chain reaction,qPCR)检测各组软骨组织IL-4、IL-6、IL-10、TNF-α及NF-κB p65的mRNA表达水平,蛋白质印迹法检测各组软骨组织NF-κB p65及p-NF-κB p65的蛋白表达水平。结果:模型组大鼠精神萎靡,毛发凌乱,饮食饮水较少;独活寄生汤各剂量组和阳性组大鼠精神状态较好,毛发饮食等恢复正常,其中高剂量和阳性组恢复最好。空白组大鼠软骨组织细胞排列整齐,结构清晰,软骨基质着色均匀;模型组大鼠细胞排列紊乱,结构不清,表面粗糙;与模型组比较,阳性组和独活寄生汤各剂量组大鼠软骨组织结构较完整、清晰,表面较光滑,软骨基质着色较均匀。与空白组比较,模型组IL-4、IL-6及TNF-α血清中含量及软骨组织mRNA表达水平均升高,IL-10水平降低(P<0.05);与空白组比较,模型组软骨组织中NF-κB p65 mRNA及蛋白水平均升高(P<0.05);与模型组比较,独活寄生汤各剂量组、阳性组的血清的IL-4、IL-6、TNF-α含量及NF-κB p65、p-NF-κB p65蛋白水平均降低(P<0.05);与模型组比较,独活寄生汤中、高剂量组、阳性组IL-4、IL-6、TNF-α及NF-κB p65软骨组织mRNA水平均降低(P<0.05);血清IL-10含量、软骨组织mRNA水平升高(P<0.05)。结论:独活寄生汤可调控IL-4、IL-6、IL-10、TNF-α、NF-κB p65及p-NF-κB p65表达,可能是治疗膝骨关节炎的机制之一。

独活寄生汤对大鼠腰椎间盘细胞活性及CaM-CaMK-CREB信号通路的影响

目的研究独活寄生汤治疗腰椎间盘突出症与Ca M-Ca MK-CREB信号通路的内在关联。方法用独活寄生汤中药血清、空白血清及胎牛血清对大鼠腰椎间盘纤维环细胞进行分组干预。采用MTT比色法检测独活寄生汤中药血清作用后的纤维环细胞生长曲线;应用western blotting技术检测独活寄生汤中药血清对Ca M、Ca MK、CREB表达及定量的影响,并对检测结果进行数据处理及统计学分析。结果中药血清干预后的第1天,5组之间两两比较无明显差异(P>0.05);中药血清干预后的第2天至第7天各组与空白血清组比较均有明显差异(P<0.05);与空白血清组比较,各剂量独活寄生汤中药血清均能可显著提高大鼠腰椎间盘纤维环细胞内的Ca M、Ca MKII、Ca MKIV和CREB等蛋白的表达水平(P<0.01),且随着中药剂量的增加Ca M、Ca MKII、Ca MKIV和CREB等蛋白的表达水平提升能力也在增加。结论独活寄生汤中药血清可明显促进大鼠椎腰间盘纤维环细胞生长,随着中药剂量的增加中药血清促进细胞生长能力增强;临床上独活寄生汤治疗腰椎间盘突出症效应过程可能存在一条重要的信号通路"Ca M-Ca MK-CREB"。

独活寄生汤联合针灸治疗对类风湿性关节炎患者红细胞沉降率、C-反应蛋白及关节功能的影响

目的探讨独活寄生汤联合针灸治疗对类风湿性关节炎患者红细胞沉降率、C-反应蛋白及关节功能影响。方法选自我院于2016年5月至2017年5月收治的类风湿关节炎患者94例分为观察组及对照组。对照组采用针灸治疗,观察组联合独活寄生汤治疗。对比2组疗效,治疗前后关节疼痛、关节肿胀、关节压痛积分,压痛数、肿胀数和VAS评分变化,及红细胞沉降率和C-反应蛋白水平变化。结果观察组总有效率(93.62%)高于对照组(74.47%)(P<0.05)。2组治疗后关节疼痛、关节肿胀、关节压痛积分均降低(P<0.05),且观察组低于对照组(P<0.05)。2组治疗后压痛数、肿胀数和VAS评分均降低(P<0.05),且观察组低于对照组(P<0.05)。2组治疗后红细胞沉降率和C-反应蛋白水平均降低(P<0.05),且观察组低于对照组(P<0.05)。结论独活寄生汤联合针灸治疗可降低类风湿性关节炎患者红细胞沉降率、C-反应蛋白,改善患者关节功能。

独活寄生汤含药血清细胞外囊泡对类风湿关节炎滑膜成纤维细胞的影响

背景:独活寄生汤是治疗类风湿关节炎的经典方剂,但具体作用机制尚不清楚,细胞外囊泡具有细胞间通讯及传递信号的强大功能,可能是该方剂发挥作用的信号载体。目的:探讨独活寄生汤含药血清细胞外囊泡对类风湿关节炎滑膜成纤维细胞增殖、迁移、侵袭及凋亡的影响。方法:肿瘤坏死因子α体外诱导构建类风湿关节炎滑膜成纤维细胞模型,实验分为5组:正常组、模型组、独活寄生汤含药血清组、独活寄生汤含药血清细胞外囊泡组、生理盐水血清细胞外囊泡组。采用CCK-8法筛选独活寄生汤含药血清及独活寄生汤含药血清细胞外囊泡作用的最佳浓度及时间;ELISA检测细胞上清炎症因子水平;划痕实验检测细胞迁移能力;Transwell实验检测细胞侵袭能力及迁移能力;Hoechst染色检测细胞凋亡情况;qRT-PCR及Western blot检测细胞凋亡相关基因及蛋白表达。结果与结论:(1)独活寄生汤含药血清作用的最佳体积分数为10%,独活寄生汤含药血清细胞外囊泡作用的最佳质量浓度为10 ng/mL,二者作用的最佳时间均为24 h;(2)与模型组相比,独活寄生汤含药血清、独活寄生汤含药血清细胞外囊泡及生理盐水血清细胞外囊泡均能抑制类风湿关节炎滑膜成纤维细胞的炎症因子表达(P<0.05)、划痕愈合(P<0.05)、迁移及侵袭(P<0.05),且独活寄生汤含药血清细胞外囊泡的抑制作用更为突出(P<0.05);(3)独活寄生汤含药血清及独活寄生汤含药血清细胞外囊泡均可促进类风湿关节炎滑膜成纤维细胞的凋亡;(4)与模型组相比,独活寄生汤含药血清、独活寄生汤含药血清细胞外囊泡及生理盐水血清细胞外囊泡均可增加促凋亡因子Caspase-3、Caspase-9、Bax表达(P<0.05),降低抑凋亡因子Bcl-2表达(P<0.05),且独活寄生汤含药血清细胞外囊泡对凋亡相关因子的调节作用更显著。上述实验结果表明独活寄生汤含药血清细胞外囊泡可以抑制类风湿关节炎滑膜成纤维细胞的过度增殖、迁移、侵袭并促进其凋亡。

独活寄生汤对CIA大鼠滑膜IL-1β、IL–8表达的影响

目的:研究中药独活寄生汤对胶原诱导性关节炎(collagen induced arthritis,CIA)大鼠滑膜白介素-1β(interleukin-1β,IL-1β)、白介素-8(interleukin-8,IL–8)表达的影响,探讨独活寄生汤治疗CIA的作用机制。方法:选用健康雄性Wistar大鼠,随机分为造模组、正常对照组,采用牛Ⅱ型胶原乳剂于大鼠尾根部及颈背部多点注射建立CIA模型。造模成功后,选取20只造模成功大鼠随机分为模型组(10只)、独活寄生汤组(简称中药组,10只)。中药组大鼠按22.9 g/kg剂量灌胃中药独活寄生汤,正常组、模型组灌胃等量生理盐水,每日1次。给药30天后,运用免疫组化法检测大鼠滑膜IL-1β、IL–8表达,并于给药前后进行关节炎指数(arthritis index,AI)评分。结果:与模型组比较,中药组治疗后大鼠关节炎指数(AI)评分明显降低(P<0.01);与治疗前比较,中药组大鼠AI评分均明显降低(P<0.01)。与正常组比较,模型组大鼠滑膜IL-1β、IL–8表达水平明显升高(P<0.01);与模型组比较,中药组大鼠滑膜IL-1β、IL–8表达水平明显降低(P<0.01)。结论:降低CIA大鼠滑膜IL-1β、IL–8表达可能是独活寄生汤治疗CIA的作用机制之一。

分子对接法预测独活寄生汤对TNF-α和IL-1β作用的活性成分

目的探讨独活寄生汤中作用于TNF-α和IL-1β受体的成分。方法采用Discovery Studio 2.0软件,通过Dock Ligands(LigandFit)模块,研究独活寄生汤中成分与受体TNF-α和IL-1β之间的相互作用,在此基础上发现潜在的活性成分。结果以TNF-α和IL-1β为靶点,与原配体的打分阈值对比,从复方中筛出与其结合较好的成分分别有10个和1个。结论独活寄生汤治疗骨性关节炎的机制,可能与其能有效降低患者关节液中IL-1β和TNF-α水平有关。

独活寄生汤加黄芪对退变人骨滑膜细胞TNF-α及IL-6mRNA的影响

目的:观察独活寄生汤加黄芪方血清对退变人骨滑膜细胞TNF-α与IL-6mRNA的影响。方法:以SPF级雄性SD大鼠制备药物血清。将P2代的退变人骨滑膜细胞分别予独活寄生汤加减方血清、莫比可血清及生理盐水血清干预24小时。通过qRT-PCR检测TNF-α、IL-6mRNA的表达。结论:独活寄生汤加黄芪方对退变人骨滑膜细胞TNF-α、IL-6的表达有明显抑制作用而发挥治疗膝骨关节炎。

独活寄生汤对大鼠类风湿关节炎的治疗效果及对血清IL-1β、TNF-α和MMPs的影响

目的研究独活寄生汤对大鼠类风湿关节炎(rheumatoid arthritis, RA)的治疗效果及对血清IL-1β、TNF-α和MMPs的影响。方法通过向雄性SD大鼠足跖面皮下注射牛Ⅱ型胶原建立大鼠胶原性关节炎(collagen-induced arthritis, CIA)模型。将雄性SD大鼠随机分为对照组(Control),模型组(Model),Model+独活寄生汤低剂量组(Model+L),Model+独活寄生汤中剂量组(Model+M),Model+独活寄生汤高剂量组(Model+H),于CIA造模21 d进行对应处理,持续36 d。测定不同实验组大鼠的体质量变化、足肿胀情况及关节炎指数评分,通过试剂盒检测抗氧化酶肝组织中SOD、CAT、GSH及氧化指标MDA活性,ELISA检测血清IL-1β、TNF-α的水平,Western Blot法分析血清MMPs的表达情况。结果 CIA模型组大鼠出现明显的RA症状,包括关节炎评分高、足肿胀、肝脏SOD、CAT、GSH活性降低及MDA活性升高,血清IL-1β、TNF-α和MMPs表达增加(均P<0.05)。独活寄生汤可呈浓度依赖性显著改善上述变化(均P<0.05),高剂量独活寄生汤对调节CIA大鼠体质量、足肿胀情况及关节炎指数评分作用明显。且血清IL-1β、TNF-α、MMP-1、MMP-3和MMP-13蛋白表达水平在高剂量处理的CIA大鼠中减少至正常大鼠相似的水平。结论独活寄生汤具有减轻RA症状、抑制促炎因子的表达及抗氧化作用,作为治疗RA的药物具有进一步研究的价值。

基于NLRP3/caspase-1信号通路影响软骨细胞焦亡探讨独活寄生汤干预类风湿关节炎作用机制

目的探讨独活寄生汤通过NLRP3/caspase-1信号通路调控软骨细胞焦亡影响类风湿关节炎(rheumatoid arthritis,RA)的作用机制。方法动物水平采用佐剂性关节炎(AA)构建RA大鼠模型,将模型大鼠随机分为两组,分别采用中药独活寄生汤和西药依托考昔干预,足趾容积测量仪检测足趾容积评估两组模型大鼠治疗前后关节肿胀情况,并采用ELISA检测治疗前后两组模型大鼠血清中类风湿因子(RF)、白细胞介素-1β(IL-1β)及白细胞介素-18(IL-18)水平。细胞水平通过胞外酸化法诱导大鼠关节软骨细胞,构建RA模型,使用独活寄生汤或依托考昔处理细胞,采用RT-qPCR及Western blotting检测各组细胞中NLRP3、caspase-1、凋亡相关斑点样蛋白(ASC)、IL-1β、IL-18 mRNA及蛋白水平,流式细胞术检测细胞焦亡水平。结果动物实验结果提示,与治疗前相比,独活寄生汤或依托考昔治疗后,RA模型大鼠关节肿胀程度均显著降低,且血清中RF、IL-1β及IL-18水平显著下降。细胞实验结果表明,独活寄生汤与依托考昔干预均可显著下调胞外酸化法诱导的RA模型细胞中NLRP3、caspase-1、ASC、IL-1β、IL-18 mRNA及蛋白水平,并显著抑制RA模型细胞焦亡。结论独活寄生汤通过抑制NLRP3/caspase-1信号通路介导的软骨细胞焦亡缓解类风湿关节炎症状是其发挥药效的途经之一。

独活寄生汤对膝骨关节炎大鼠IL-17/NF-κB通路的影响

目的:探讨独活寄生汤通过调控IL-17/NF-κB信号通路治疗膝骨关节炎(KOA)的作用机制。方法:将大鼠随机分为正常组、模型组、独活寄生汤组和双醋瑞因组,采用关节腔注射碘乙酸钠溶液建立KOA模型。确认造模成功后,分别采用药物干预30 d,观察大鼠膝关节软骨病理形态变化。酶联免疫吸附试验(ELISA)检测大鼠血清中白细胞介素17(IL-17)和肿瘤坏死因子α(TNF-α)的表达。Real-time PCR和免疫组化法检测软骨核因子κB p65(NF-κB p65)、肿瘤坏死因子受体相关因子6(TRAF6)、NF-κB激活剂1(ACT1)mRNA和蛋白表达水平。结果:独活寄生汤组大鼠膝关节软骨表面稍不平整,软骨细胞数量减少,排列较规则,软骨基质轻染。与正常组比较,模型组大鼠血清IL-17、TNF-α表达水平升高(均P<0.01)。与模型组比较,独活寄生汤组大鼠血清IL-17、TNF-α表达下降(均P<0.05)。与正常组比较,模型组TRAF6 mRNA表达升高,NF-κB p65、ACT1 mRNA表达升高(均P<0.01)。与模型组比较,独活寄生汤组NF-κB p65、ACT1、TRAF6 mRNA表达降低(均P<0.05)。与正常组比较,模型组大鼠软骨NF-κB p65、TRAF6和ACT1蛋白表达水平升高(均P<0.01)。与模型组比较,独活寄生汤组大鼠软骨NF-κB p65、TRAF6、ACT1的蛋白表达降低(均P<0.01)。结论:独活寄生汤通过调节IL-17/NF-κB信号通路,降低IL-17、TNF-α促炎因子表达,有效缓解KOA模型大鼠症状。