Introduction Melittin, a 26-amino acid polypeptide with various pharmacological effects, possesses potential anti-tumor properties. The present study examined the possible mechanisms of action on Osteosarcoma (OS) cells.Materials and Methods In the beginning, the proliferation of human osteoblast precursor cell line hFOB 1.19 was assessed. A series of in-vitro tests was implemented to investigate the effects of melittin on the survival, apoptosis, migration, and invasion of OS cells (143-B and MG63). An immunoblotting assay was applied to quantify the protein expression of potential mediators. Further, the mechanisms of action of melittin on OS cells were validated through rescue assays.Results In OS cells 143-B and MG63, melittin treatment evidently reduced the cell viability, migration, and invasion and elevated apoptosis, with downregulated Bcl-2 and upregulated Bax and cleaved caspase-3. Moreover, the phosphorylation level of PI3K/AKT/mTOR was diminished following melittin treatment. In contrast, the rescue assay demonstrated that the anti-OS effects of melittin in vitro were negated by the 740 Y-P (a known PI3K activator). This was evidenced by altered viability, migration, and invasion, reduced apoptosis in OS cells, and the expression of apoptosis-related mediators.Discussion Utilizing an in-vitro OS cell model, the current study discovered that melittin can suppress the growth, migration, and invasion of OS cells yet promote apoptosis, potentially via targeting PI3K/AKT/mTOR pathway, providing a novel mechanism underlying the effect of melittin on OS cells.Conclusion The research demonstrated the anti-OS effects of melittin and its possible relationship with the PI3K/AKT/mTOR pathway.

OBJECTIVE:The objective of this study is to analyze and verify the main drug components and targets of "Fuzi-Guizhi" in the treatment of osteoarthritis by using the network pharmacology platform.METHODS:The integrated pharmacology of "Fuzi-Guizhi" was analyzed by using the platform of integrated pharmacology of traditional Chinese medicine to explore its mechanism in the treatment of osteoarthritis. By establishing an arthritis model in vitro, the pharmacological effect of "aconitecassia twigs" on articular cartilage was evaluated and conducted for molecular docking.RESULTS:28 candidate active components, 37 compound targets, and 583 osteoarthritis-related potential targets were screened, and 10 key target processes were screened in the protein interaction network model. Enrichment analysis showed that the 10 core targets involved 958 GO biologic function items and 76 KEGG signal pathways, which were mainly related to apoptosis and mitochondrial functional metabolism and "Fuzi-Guizhi" drug-containing serum inhibited the expression of Caspase-3 mRNA and protein in chondrocytes and promoted the synthesis of ATP.CONCLUSION:Our research is preliminary that the mechanism of action of "Fuzi-Guizhi" may inhibit chondrocyte degeneration by resisting mitochondrial apoptosis, and further experimental research is required to determine.

Acute ankle sprain is one of the most common lower limb injuries particularly in athletes and it accounts for 16-40% of all sports-related injuries. A significant proportion of patients experience persistent residual symptoms and recurrence. Traditional Chinese medicine (TCM) offers effective treatment in terms of pain relief, swelling reduction, functional improvement, and shorter disease duration. This study provides a comprehensive analysis of the active ingredients, molecular targets, signaling pathways, and Longji Xiaozhong ointment's clinical efficacy for treating acute ankle sprains. We followed network pharmacology approach to identify the molecular targets and mechanism of action of Longji Xiaozhong ointment in treating acute ankle sprain and to validate its clinical value using relevant clinical data. Initially, Longji Xiaozhong Ointment's active ingredients were screened using the TCMSP platform based on drug properties similar to DLP >0.18 and oral bioavailability OB >= 30%. The molecular targets were identified from the ETCM database, followed by constructing protein-protein interaction (PPI) network diagrams and GO and pathway enrichment analysis on key genes. Furthermore, the UniProt database was used to investigate the 3D structures of these key genes. Finally, the clinical data of 111 patients with acute ankle sprains were retrospectively analyzed and further explored by comparing the different curative effects of Longji Xiaozhong ointment with Celebrex and Shexiang Analgesic ointment to check the clinical value of Longji Xiaozhong ointment in treating acute ankle sprain. In the TCMSP database, we found that there were 42 active ingredients in Longji Xiaozhong ointment, including 13 kinds of Strychnosnux-vomica L., 7 kinds of Ligusticum chuanxiong, and 22 kinds of Safflowers (Carthamus tinctorius), and a total of 3206 target genes were obtained. After applying the OB >= 50% criteriaand DL >0.5, 19 key target genes were selected based on their correspondence to the active ingredients. Protein mutual aid network construction and module analysis yielded two high-scoring Clusters and identified seven key proteins, including SLC6A4, ADRB2, ADRA1B, CHRM1, F2, OPRM1, and OPRD1. Functional enrichment analysis (FEA) of candidate target genes of Strychnosnux-vomica, Ligusticum chuanxiongand Safflowers in Longji Xiaozhong ointment in ETCM database showed SUMOylation of intracellular receptors, blockade of NMDA receptors, and activation of GABA A receptors, lipid metabolism regulationby peroxisome proliferator-activated receptor alpha (PPAR alpha). PPARA activates aspects of gene expression and transcriptional activation of mitochondrial biogenesis. On retrospective analysis, all patients were divided into Group A: Celebrex combined with brace immobilization group (36 cases); Group B: Shexiang Jietong ointment combined with brace immobilization group (37 cases); and Group C: Longji Xiaozhong ointment combined with brace immobilization group (38 cases). After treatment, the VAS score, swelling degree and scope score of Longji Xiaozhong ointment combined with brace immobilization group were significantly lower than those of the other two groups, and the effect was significantly better in terms of daily activities, sports, quality of life and occurrence of re-sprain than the other two groups. Our findings, supported by clinical data, demonstrate that Longji Xiaozhong ointment effectively alleviates swelling and pain, accelerates the repair of ankle ligaments, enhances ankle joint function, and improves ankle joint stability.

目的:观察电针腰椎华佗夹脊穴与椎间孔镜手术治疗腰椎间盘突出症(LDH)临床疗效.方法:收集187例LDH患者,根据患者意愿分为电针组(n=95)和椎间孔镜组(n=92).电针组在病变节段患侧选取华佗夹脊穴施以电针治疗,椎间孔镜组则在椎间孔镜系统下行病变节段突出髓核摘除术.比较两组患者VAS、ODI、SF-36量表评分及直腿抬高角度、疗效.结果:治疗后,两组患者VAS、ODI、SF-36评分和直腿抬高角度较治疗前改善(均P<0.05).治疗后1个月、治疗后3个月,椎间孔镜组VAS、ODI、SF-36评分及直腿抬高角度改善优于电针组(均P<0.05).治疗后6个月,两组患者VAS、ODI评分比较,差异无统计学意义(均P>0.05),椎间孔镜组直腿抬高角度、SF-36评分改善优于电针组(均P<0.05).治疗后12个月,两组患者VAS、ODI、SF-36评分及直腿抬高试验比较,无统计学意义(均P>0.05).治疗后1个月,椎间孔镜组优良率高于电针组(P<0.05);治疗后12个月,椎间孔镜组优良率与电针组比较,差异无统计学意义(P>0.05).结论:电针腰椎华佗夹脊穴与椎间孔镜手术治疗腰椎间盘突出症均有效,可改善临床症状,提高生活质量.椎间孔镜组近期疗效优于电针治疗组,两组远期疗效相近.

Background and Purpose: Osteosarcoma is the most commonly seen type of primary malignant bone tumors in children and adolescents. Partial patients with osteosarcoma cannot tolerate the side effects of chemotherapy drugs. Hence, it is urgent to find anti-osteosarcoma drugs with low side effects. Melittin is an anti-tumor Traditional Chinese Medicine with low side effects. The purpose of this study was to explore the anti-osteosarcoma effect of melittin and its possible molecular mechanisms. Methods: The effects of melittin on cell growth were detected by CCK-8, clonal formation, and flow cytometry. The related molecules were also investigated by Real-time PCR and Western blot. A xenograft model in nude mice was established to observe the effects of melittin on tumor growth and the related molecular expression was detected by immunohistochemistry. Results: Melittin can inhibit the proliferation of osteosarcoma 143B cells, reduce colony formation, and induce apoptosis while significantly up-regulating the expression of Bax and Caspase-3 and down-regulating the expression of Bcl-2 proteins. Moreover, treatment with melittin significantly reduced the mRNA and protein levels of β-catenin and Wnt/β- catenin related genes (LRP5, c-Myc, and Survivin) in osteosarcoma 143B cells in vitro. The xenograft model found that melittin significantly inhibited tumor growth and decreased the protein expression levels of β-catenin and Wnt/β- catenin related genes in vivo. Conclusion: These findings show that melittin could inhibit the growth of osteosarcoma 143B cells, which may be related to the inhibition of Wnt/β-catenin signaling pathway activity and induce apoptosis by up-regulating the ratio of Bax/Bcl-2 in osteosarcoma 143B cells. Therefore, melittin is a promising anti-tumor drug for the treatment of osteosarcoma.

OBJECTIVEThis study aims to investigate the regulatory role of exosome lncRNA OIP5-AS1 in tumor progression and autophagy.METHODSSeventy-three cases of osteosarcoma (OS) tissues and 56 cases of adjacent normal tissues were collected to culture human OS cell line HOS. The exosomes secreted by OS cell line were isolated and collected. Apoptosis and exosome markers were detected by flow cytometry. A nude mouse model of OS was established. The gene expression levels of lncRNA OIP5-AS1, miR-153 and autophagy-related protein 5 (ATG5) were quantified by real-time quantitative PCR (RT-PCR). The binding sites of lncRNA OIP5-AS1 and miR-153 were predicted by Starbase3.0, and the binding sites of miR-153 and ATG5 were predicted by Targetscan7.2. The gene binding sites were verified by luciferase reporter gene detection or RNA immunoprecipitation (RIP). The relative level of protein was tested by Western blot. Transwell was applied to test migration and invasion of OS cells. The angiogenesis of OS cells was tested by tubule formation test.RESULTSThe results of RT-PCR showed that lncRNA OIP5-AS1 levels were elevated in OS cells and exosomes secreted by cells. Cell function experiments revealed that the proliferation, migration, and invasion of OS cells were promoted by exosomal lncRNA OIP5-AS1. In exosomes, lncRNA OIP5-AS1 inhibited the expression of LC3-II and Beclin 1 proteins, indicating that exosomal lncRNA OIP5-AS1 inhibited autophagy. According to the results of bioinformatics tools and dual-luciferase reporter (DLR) assay or RNA immunoprecipitation (RIP), miR-153 targeted the 3'-UTR of lncRNA OIP5-AS1 and autophagy-related protein 5 (ATG5). The results of western blot (WB) assay showed that exosomal lncRNA OIP5-AS1 and down-regulated miR-153 led to the enhancement of ATG5 protein expression, while up-regulated miR-153 resulted in the decrease of ATG5 protein expression. ATG5 was negatively correlated with miR-153 and positively correlated with lncRNA OIP5-AS1. The results of tubule formation assay disclosed an increase in the angiogenesis level caused by the exosomal lncRNA OIP5-AS1, which was then reversed by the increase of miR-153 and decrease of ATG5.CONCLUSIONHighly enriched exosomal lncRNA OIP5-AS1 can regulate OS tumor angiogenesis and autophagy through miR-153 and ATG5.

Melittin is a major active peptide component of bee venom that has been demonstrated to show anti-tumor effects. Osteosarcoma is a type of bone tumor with a high degree of malignancy, and metastasis is the main challenge of osteosarcoma therapy. This study aimed to investigate the role of melittin in the lung metastasis of osteosarcoma. 143 B cells were treated with different concentrations of melittin in vitro. Wound-healing and transwell assays were performed to determine the cell migration and invasion potential. Quantitative real-time PCR and Western blot experiments were performed to evaluate the expression levels of Wnt/β-catenin signaling pathway-related factors after treatment with melittin. The orthotopic implantation model and hematoxylin-eosin staining were used to investigate the effect of melittin treatment on tumor formation and lung metastasis. Immunohistochemical staining and Western blot experiments were performed to indicate the melittin-mediated expression changes in Wnt/β-catenin signaling pathway-related factors. The cell migration and invasion potential were observed to be inhibited in a dose-dependent manner upon treatment with melittin. Treatment with medium and high concentrations of melittin attenuated the mRNA and protein expression of LRP5, β-catenin, MMP-2, cyclin D, c-Myc, survivin, MMP-9, and VEGF genes in vitro. Melittin significantly inhibited the growth of tibia xenografts in nude mice and decreased the number of lung metastatic nodules. Consistent with the results observed in vitro, treatment with melittin at medium and high concentrations attenuated the expression of Wnt/β-catenin signaling pathway-related factors in vivo. In vitro, Wnt/β-catenin signaling pathway was involved in Melittin-mediated -migration and invasion potential of 143 B cells. Similarly, as observed in the in vivo experiments, Wnt/β-catenin signaling pathway was also associated with the role of melittin on lung metastasis of osteosarcomas.

甲状腺癌是内分泌系统最常见的肿瘤之一.环状RNA(circular RNA,circRNA)是一种高度保守的小分子非编码RNA.近年研究发现,多种circRNA在不同类型甲状腺癌中表达异常,可能在甲状腺癌的发生和发展过程中有重要作用.本文从分子生物学特性综述circRNA在甲状腺癌发生、发展、诊治和预后评估中的研究现状.

目的 观察补中益气汤对原发性肝癌患者肝动脉化疗栓塞术(TACE)后疲劳指数及生活质量的影响,为临床治疗提供参考依据.方法 选取我院收治的60例肝动脉化疗栓塞术后患者为研究对象,随机分为治疗组和对照组各30例,治疗组术后在常规治疗基础上加服补中益气汤煎剂,对照组术后采取常规对症治疗,观察8周后两组的疲劳评分及生活质量评分的比较.结果 治疗组在术后第4周、第8周简明疲劳量表(BFI)指数较治疗前持续下降(P＜0.05),且第8周与对照组同期比较有统计学意义(P＜0.05);对照组BFI指数虽也呈下降趋势,但差异不明显(P＞0.05);治疗组与对照组的肝胆疾病特异量表(FACT-Hep)中躯体日常功能(PWB)、社交/家庭生活(SWB)、功能状况(FWB)及肝胆疾病特异模块(HEP)四个维度和总分均较治疗前明显升高(P＜0.05),其中治疗组在PWB、SWB、FWB三-个维度和总分的治疗后评分与对照组同期相比,差异有统计学意义(P＜0.05),HEP与对照组同期无显著差异(P＞0.05),而情感状况(EWB)维度治疗前后无明显变化(P＞0.05).结论 补中益气汤可有效改善患者肝动脉化疗栓塞术后导致的疲劳,提高患者生活质量,值得临床推广.

This study aimed to identify co-expressed differentially expressed genes (DEGs) in quiescence and senescence of osteosarcoma (OS) U2OS cells and investigate their biological functions. GSE94805 from Gene Expression Omnibus database was extracted, involving 12 samples of OS U2OS cells (4 quiescence, 4 senescence, and 4 control samples). After analysis of DEGs by limma package, VENN analysis was performed to identify co-expressed DEGs in quiescence and senescent. The Cytoscape software was used to construct an interactive network of co-expressed DEGs. Finally, box-plot was drawn for the co-expressed DEGs in sub-network. Besides, the relation literatures were selected in GenCLiP database for the co-expressed DEGs. Seven hundred and forty-three DEGs (255 up-regulated genes, 488 down-regulated genes) were obtained in quiescence and 2135 DEGs (1189 up-regulated genes, 946 down-regulated genes) in senescence. Through VENN analysis, 448 DEGs (131 up-regulated genes, 317 down-regulated genes) were co-expressed in quiescent and senescence. In the co-expressed DEGs network, 896 nodes (448 nodes in quiescent, 448 nodes in senescent) were obtained. Finally, 16 co-expressed DEGs were obtained in the sub-network analysis, in which Aurora kinase A (AURKA) and polo-like kinase (PLK4) had been reported in OS. AURKA and PLK4 might be the key genes in quiescence and senescence of OS U2OS cells.

Background: β-catenin plays a crucial role in the progression of osteosarcoma. However, the clinical significance of β-catenin over-expression in osteosarcoma still remains unclear. Thus, we performed a meta-analysis of studies that evaluated the impact of β-catenin on metastasis and overall survival (OS) in osteosarcoma. Methods: We searched PubMed, The Cochrane Library, Embase, Springer, Science Direct, OVID, Weipu, Wanfang and China National Knowledge Internet (CNKI) databases from their start year up to Aug.2019. Individual hazard ratios (HRs) and 95% confidence intervals (CIs) were extracted and pooled HRs with 95% CIs or odd ratio (OR) were used to evaluate the relationships between β-catenin over-expression and metastasis and overall survival in osteosarcoma. Results: Eight related studies involving 521 patients were qualified for this meta-analysis. Results showed that over-expression of β-catenin was significantly correlated with metastasis (OR = 3.31, 95% CI = 2.08–5.24, P < 0.001) and overall survival (HR = 2.32, 95% CI = 1.48–363, P = 0.02). Conclusion: The meta-analysis revealed that over-expression of β-catenin might be associated with distant metastasis and overall survival in osteosarcoma, which reminds that β-catenin acts as a prognostic biomarker and it can guide the clinical therapy in osteosarcoma patients.

Wnt/β-catenin信号通路是调节细胞发育和生长的一个关键途径[1].在正常情况下对胚胎发育和器官的形成和功能起着重要的调节作用,此通路的失调参与人类多种肿瘤的发病过程[2].骨肉瘤是儿童和青少年长骨干骺端常见的原发性恶性骨肿瘤,以产生骨样基质为病理特征,具有高度异质性、高复发率、多药耐药性、早期易肺转移等特点[3].以外科手术联合甲氨蝶呤、顺铂等新辅助化疗的应用,使骨肉瘤患者的 5 年生存率提高到 75％ 左右[4] ,但却不能有效地降低总病死率和改善整体预后,出现转移或复发的患者5 年生存率 <20％ [5].研究表明Wnt/β-catenin 信号通路参与骨肉瘤细胞的生长、侵袭和转移,并有效诱导骨肉瘤的耐药和再生,是介导骨肉瘤发生和发展的一个重要因素[6].

Wnt/β-catenin信号通路在调控细胞增殖、分化中起重要作用,其活性异常表达能引起肿瘤的发生,被认为是肿瘤发生过程中的关键信号通路.骨肉瘤是儿童及青少年常见的原发性恶性骨肿瘤.以往的研究证实Wnt/β-catenin信号异常激活是导致骨肉瘤发生和发展的重要因素之一.体内及体外实验研究中通过抑制Wnt/β-catenin信号通路的活性可以抑制骨肉瘤细胞的生长和侵袭转移.因此进一步探索Wnt/β-catenin信号通路抑制剂可能成为骨肉瘤治疗的新热点.本文将近5年来关于Wnt/β-catenin信号通路抑制剂在骨肉瘤中的研究做一综述.

类风湿性关节炎是临床常见的自身免疫性疾病,治疗不当易导敛关节畸形、功能丧失,影响患者日常生活.中医药在治疗类风湿疾病中具有独特的优势,附子作为具有抗炎、抗寒冷、免疫调节作用的中草药,其复方在阳的治疗上具有明显的疗效,能够改善症状,但作用机理不明确.文章结合近5年有关附子复方对类风湿关节炎动物模型的实验研究进行综述,为附子复方在类风湿关节炎的防治起到一定的参考作用.

部分骨肉瘤患者对化疗缺乏有效反应,其耐药机理目前尚不明确.骨肉瘤的难治性、复发转移和抗药性与癌干细胞有关.癌干细胞可通过细胞表面特异性标记物、侧群细胞、细胞球形成等进行辨别.多项研究发现,骨肉瘤中存在Wnt/β-catenin信号转导通路的异常表达.通过调控该信号转导通路可改变骨肉瘤癌干细胞的特性,抑制骨肉瘤的增殖、复发和远处转移.该文对Wnt/β-catenin信号转导通路对骨肉瘤癌干细胞的调节作用及治疗前景作一综述.

Endothelial progenitor cells (EPCs) are important in tumor angiogenesis. Stromal cell-derived factor-1 alpha (SDF-1 alpha) and its receptor C-X-C chemokine receptor type 4 (CXCR4) are key in stem cell homing. Melittin, a component of bee venom, exerts antitumor activity, however, the underlying mechanisms remain to be elucidated. The present study aimed to assess the effects of melittin on EPCs and angiogenesis in a mouse model of osteosarcoma. UMR-106 cells and EPCs were treated with various concentrations of melittin and cell viability was determined using the MTT assay. EPC adherence, migration and tube forming ability were assessed. Furthermore, SDF-1 alpha, AKT and extracellular signal-regulated kinase (ERK) 1/2 expression levels were detected by western blotting. Nude mice were inoculated with UMR-106 cells to establish an osteosarcoma mouse model. The tumors were injected with melittin, and its effects were assessed by immunohistochemistry and immunofluorescence. Melittin decreased the viability of UMR-106 cells and EPCs. In addition, it decreased EPC adhesion, migration and tube formation when compared with control and SDF-1 alpha-treated cells. Melittin decreased the expression of phosphorylated (p)-AKT, p-ERK1/2, SDF-1 alpha and CXCR4 in UMR-106 cells and EPCs when compared with the control. The proportions of cluster of differentiation (CD) 34/CD133 double-positive cells were 16.4 +/- 10.4% in the control, and 7.0 +/- 4.4, 2.9 +/- 1.2 and 1.3 +/- 0.3% day, respectively (P<0.05). At 11 days, melittin reduced the tumor size when compared with that of the control (control, 4.8 +/- 1.3 cm(3); melittin, 3.2 +/- 0.6, 2.6 +/- 0.5, and 2.0 +/- 0.2 cm(3) for 160, 320 and 640 mu g/kg, respectively; all P<0.05). Melittin decreased the microvessel density, and SDF-1 alpha and CXCR4 protein expression levels in the tumors. Melittin may decrease the effect of osteosarcoma on EPC-mediated angiogenesis, possibly via inhibition of the SDF-1 alpha/CXCR4 signaling pathway.

目的 检测成骨-破骨细胞共育体系中应用加味三根汤含药血清后抗酒石酸酸性磷酸酶(TRACP)以及白细胞介素-1β(IL-1β)、肿瘤坏死因子(TNF-α)的变化,探讨加味三根汤的作用机制.方法 建立成骨-破骨细胞共育体系,制作SD大鼠含药血清,分为空白对照组,加味三根汤低、中、高剂量组,每组设6个复孔,采用酶联免疫吸附试验(ELISA)检测共培养体系上清液中TRACP及IL-1β、TNF-α的含量.结果 经鉴定成功培养成骨细胞和破骨细胞,并建立成骨-破骨细胞共育体系.加味三根汤含药血清中、高剂量组TRACP活性低于空白血清对照组,差异具有统计学意义(P＜0.05).与空白血清对照组比较,加味三根汤中、高剂量组IL-1β含量有所下降,加味三根汤高剂量组TNF-α含量降低,差异具有统计学意义(P＜0.05).结论 加味三根汤能够抑制共育体系中破骨细胞的功能,此作用可能是通过抑制IL-1β、TNF-α的分泌而达到.

目的 观察蜂毒素对内皮祖细胞微血管形成的影响及探讨其作用机制.方法 通过MTT比色法、黏附实验、Transwell小室迁移实验、小管形成实验等分别测定蜂毒素对内皮祖细胞的增殖、黏附、迁移、小管形成能力的影响.并且通过Western blot法测定蜂毒素对内皮祖细胞VEGFR1、VEGFR2、AKT、ERK1/2磷酸化的影响.结果 随着蜂毒素的作用浓度增大,其对内皮祖细胞增殖的抑制力越强,蜂毒素浓度在2 μg/ml以下时,不表现出明显的细胞毒性;与0μg/ml组相比较,蜂毒素1和2 μg/ml浓度组对内皮祖细胞黏附、迁移和小管形成具有明显的抑制作用(P＜0.05);蜂毒素可抑制内皮祖细胞磷酸化作用,并且下调VEGFR2的表达,VEGFR1的表达无明显变化.结论 蜂毒素能明显抑制大鼠骨髓来源的内皮祖细胞的增殖、黏附、迁移、小管形成的能力,其作用机制可能与蜂毒素抑制内皮祖细胞的磷酸化信号转导通路,从而下调VEGFR2蛋白表达.

Objective:To observe the effect of Qianghuo Dihuang Decoction on the serum levels of tumor necrosis factor alpha(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)in rats with Freund's complete adjuvant-induced rheumatoid arthritis,and to explore the possible mechanism of Qianghuo Dihuang Decoction in the treatment of rheumatoid arthritis, so as to provide experimental basis for the application of Qianghuo Dihuang Decoction in clinic.Methods:The adjuvant arthritis rat model was established.SD rats were randomly divided into normal control group,model group,Qianghuo Dihuang Decoction group and methotrexate group.Intragastric administration method was adopted in the treatment.Enzymelinked immunosorbent assay was used for detection of the serum levels of TNF-α,IL-1βand IL-6in AA rats.Results:The serum levels of TNF-α,IL-1βand IL-6in model group were higher than those in normal control group(P0.01).After treatment,the serum levels of TNF-α,IL-1βand IL-6in Qianghuo Dihuang Decoction group and methotrexate group were lower than those in the model group(P0.01).There was no significant difference between Qianghuo Dihuang Decoction group and the methotrexate group in the serum levels of TNF-α,IL-1βand IL-6(P ﹥0.05).Conclusion:Qianghuo Dihuang Decoction can inhibit the abnormal secretion of TNF-α,IL-1βand IL-6in AA rats,so that it can prevent the development of immune inflammation in AA rats.Our experimental results showed that the effect of Qianghuo Dihuang Decoction on serum levels of TNF-α,IL-1βand IL-6is the possible mechanism in the treatment of RA.

Osteoarthritis (OA) is a degenerative joint disease and a major cause of pain and disability in older adults. We have previously identified epidermal growth factor receptor (EGFR) signaling as an important regulator of cartilage matrix degradation during epiphyseal cartilage development. To study its function in OA progression, we performed surgical destabilization of the medial meniscus (DMM) to induce OA in two mouse models with reduced EGFR activity, one with genetic modification ( Egfr Wa5/+ mice) and the other one with pharmacological inhibition (gefitinib treatment). Histological analyses and scoring at 3 months post-surgery revealed increased cartilage destruction and accelerated OA progression in both mouse models. TUNEL staining demonstrated that EGFR signaling protects chondrocytes from OA-induced apoptosis, which was further confirmed in primary chondrocyte culture. Immunohistochemistry showed increased aggrecan degradation in these mouse models, which coincides with elevated amounts of ADAMTS5 and matrix metalloproteinase 13 (MMP13), the principle proteinases responsible for aggrecan degradation, in the articular cartilage after DMM surgery. Furthermore, hypoxia-inducible factor 2α (HIF2α), a critical catabolic transcription factor stimulating MMP13 expression during OA, was also upregulated in mice with reduced EGFR signaling. Taken together, our findings demonstrate a primarily protective role of EGFR during OA progression by regulating chondrocyte survival and cartilage degradation.