Sixty Sprague-Dawley female rats were randomly divided into sham-operated groups and five ovariectomy (OVX) subgroups. Rats subjected to sham and OVX were treated with the vehicle, alendronate, and Zuogui Wan (ZGW) at the doses of low, medium and high lyophilized powder daily for 3 months, respectively. The gene or protein expression of NK1R, PPAR γ, and OSX were assayed by either quantitative polymerase chain reaction or Western blot analysis. The results showed that compared with the OVX group, ZGW could reduce the level of PPARγ and increase the levels of OSX and. Meanwhile, ZGW could prevent bone loss. In addition, we found ZGW upregulated for the NK1R mRNA or protein expression by promoting the expression level of transcription factor FoxO3 and increasing its binding to the NK1R promoter region -700/-200 sequence. These results suggest that the regulation of FoxO3 and NK1R played a role and contributed to the mechanism of ZGW underlying the increase in bone mass in the OVX rat model.

目的:系统评价柴胡疏肝散合四君子汤加减方治疗慢性萎缩性胃炎(chronic atrophic gastritis,CAG)的临床效果.方法:电子计算机检索中国知网(CNKI)、万方数据库(WANFANG DATA)、中国生物医学文献数据库(CBM)、中国科技期刊全文数据库(VIP)、Cochrane Library、Pubmed、Embase数据库,检索时间限定为建库至2020年8月17日,纳入所有柴胡疏肝散合四君子汤加减方治疗CAG的随机对照试验(randomized controlled trial,RCT),并运用RevMan5.3软件进行Meta分析.结果:最终纳入7项研究共606例CAG患者,Meta分析结果显示与对照组常规西药治疗方案比较,柴胡疏肝散合四君子汤加减方可提高CAG患者的临床总有效率[RR=1.23,95％CI(1.16,1.31),P<0.00001],降低CAG不良反应发生率[RR=0.18,95％CI(0.05,0.58),P=0.004]和CAG的复发率[RR=0.30,95％CI(0.13,0.70),P=0.005],差异有统计学意义;分层分析显示对照组吗丁啉、奥美拉唑、硫糖铝混悬液3种药物联合使用治疗CAG有效率高于其他2种药物合用(P<0.05),维酶素、果胶铋2种药物联合使用治疗CAG疗效最差(P<0.05);漏斗图分析结果显示不对称可能存在发表偏倚.结论:基于纳入7项研究的Meta分析结果,柴胡疏肝散合四君子汤加减方治疗CAG的临床疗效优于常规西药.

目的 探讨左归丸(Zuoguiwan,ZGW)的补肾填精、益髓壮骨的作用,促进骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)的增殖和向成骨细胞方向分化.方法 制备ZGW含药血清,设置10％FBS DMEM组、成骨诱导液组、2.5％空白大鼠血清-成骨诱导液组、2.5％ZGW含药血清-成骨诱导液组.采用MTT法观察各组对BMSCs的增殖,使用碱性磷酸酶钙钴染色法检测BMSCs向成骨细胞方向的分化能力,碱性磷酸酶(alkaline phosphatase,AKP)法检测细胞培养液中AKP的含量,Western blot检测BMSCs中Runt相关转录因子2(Runx2)、骨钙蛋白(osteocalcin,OPN)和碱性磷酸酶(alkaline phosphatase,ALP)蛋白的表达.结果 2.5％ZGW含药血清-成骨诱导液组能够促进BMSCs的快速增殖,较早地进入平台期,促进BMSCs中AKP的分泌,提高ALP含量,上调BMSCs细胞中Runx2、ALP和OPN的蛋白表达水平.结论 ZGW抗骨质疏松的分子机制,可能与调节Runx2激活ALP、OPN蛋白表达,促进BMSCs增殖和向成骨细胞方向分化有关.

目的 探讨CCK-8与MTT法在检测左归丸含药血清干预大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)增殖时所需的最佳实验条件,并比较两种方法的优缺点.方法 用左归丸含药血清进行干预P3 BMSCs,孵育结束后,分别加入CCK-8与MTT试剂,检测两种方法的最佳波长、培养时间、灵敏度和增殖时间.结果 CCK-8法与MTT法检测BMSCs的最佳波长分别为450 nm和550 nm,孵育时间分别为3 h和4 h,细胞接种密度分别为0.1×104/孔和0.25×104/孔.CCK-8法与MTT法在BMSCs接种密度分别高于0.25×104/孔和0.5×104/孔时,两种方法检测所得OD值均提早进入平台期,易出现假S型增长曲线.而当密度分别低于0.05×104/孔和0.1×104/孔时,细胞增加缓慢,难以形成S型增长曲线,不能反映出BMSCs的增殖特性.与CCK-8法相比较,在细胞密度低于0.1×104/孔或高于2×104/孔时,MTT法对BMSCs在翻倍时OD值增加的响应量无明显变化,差异无统计学意义.而细胞密度为0.25×104/孔至1×104/孔时,MTT法对BMSCs在翻倍时OD值增加的响应量明显增加,差异有统计学意义(P<0.05).结论 MTT法更适用于左归丸含药血清干预BMSCs增殖和分化的研究.

目的 探讨左归丸治疗绝经后骨质疏松症合并骨关节炎的可能作用机制.方法 60只雌性SD大鼠按随机数字表法分为假手术组,模型组,阿仑膦酸钠组及左归丸低、中、高剂量组,每组10只.除假手术组外,其余各组大鼠用去卵巢法建立绝经后骨质疏松症合并骨关节炎模型.造模2周后,假手术组、模型组给予蒸馏水1 ml/(kg·d)灌胃;阿仑膦酸钠组给予阿仑膦酸钠混悬液63 mg/(kg·d)灌胃;左归丸低、中、高剂量组分别给予2.42、4.84、9.68g/(kg·d)左归丸灌胃,每日1次,每周6次.12周后观察大鼠软骨组织形态学变化,检测左侧股骨远端软骨下骨骨密度(BMD),血清肿瘤坏死因子α(TNF-α)和白细胞介素1β(IL-1β)含量,股骨中P物质(SP)和Runt相关转录因子2(Runx2)蛋白表达,下丘脑SP及其受体神经激肽1受体(NK1-R)蛋白及mRNA表达.结果 与假手术组比较,病理显示模型组大鼠软骨表面破坏、有裂隙,软骨细胞排列混乱;软骨下骨BMD降低,血清TNF-α和IL-1 β水平升高,下丘脑SP和NK1-R蛋白及mRNA表达水平下调(P＜0.05或P＜0.01),Runx2、SP表达量减少.与模型组比较,病理显示阿仑膦酸钠组及左归丸中、高剂量组大鼠软骨损伤明显改善;软骨下骨BMD明显升高,血清TNF-α和IL-1β水平显著降低,下丘脑SP和NK1-R蛋白及mRNA表达水平上调(P＜0.01);阿仑膦酸钠组及左归丸低剂量组Runx2、SP表达量增多.与左归丸低剂量组比较,左归丸中、高剂量组大鼠下丘脑NK1-R mRNA表达显著升高(P＜0.05或P＜0.01).左归丸高剂量组IL-1β水平显著低于左归丸低、中剂量组(P＜0.01).结论 左归丸可能通过上调下丘脑神经肽SP及其受体蛋白、mRNA水平,提高软骨下骨骨密度,进而纠正骨代谢失衡,是其防治绝经后骨质疏松症合并骨关节炎的作用机制之一.

目的:探讨左归丸及其拆方治疗绝经后骨质疏松症(PMOP)的作用机制.方法:去卵巢法建立骨质疏松症模型,将40只SD大鼠平均随机分为假手术组,模型组(0.1 mL·kg-1·d-1),17β-雌二醇组(50 μg·kg-1·d-1)和左归丸补阳药低、高剂量(0.55,1.1 g·kg-1·d-1)组.灌胃给药12周后,显微CT(μ-CT)检测右侧远端股骨骨密度(BMD)及显微结构;苏木精-伊红(HE)染色观察大鼠股骨形态学变化;酶联免疫吸附法(ELISA)检测大鼠血清中Ⅰ型胶原交联羧基端肽(β-CTX)和骨特异性碱性磷酸酶(BALP)浓度;实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测大鼠下丘脑β2-肾上腺素能受体(β2AR),胫骨平台骨保护素(OPG)和核转录因子-κB受体激活因子配体(RANKL)蛋白及mRNA表达水平.结果:与假手术组相比,模型组大鼠BMD显著下降(P＜0.01),骨组织形态破坏;血清BALP含量降低,β-CTX含量升高(P＜0.01);下丘脑β2AR,胫骨RANKL mRNA及蛋白表达量升高(P＜0.05,P＜0.01),胫骨OPG mRNA及蛋白表达量降低(P＜0.05,P＜0.01).治疗后,与模型组比较,各用药组大鼠BMD升高(P＜0.01),骨组织形态改善;血清BALP含量升高,β-CTX含量降低(P＜0.01);下丘脑β2AR,胫骨RANKL mRNA及蛋白表达降低(P＜0.05,P＜0.01),胫骨平台OPG mRNA和蛋白表达升高(P＜0.05,P＜0.01).结论:左归丸拆方中补阳药能通过调节β2AR介导的OPG/RANKL信号通路提高去卵巢大鼠骨密度,“阳中求阴”配伍特点是左归丸治疗PMOP的关键之一.

Background: Osteoarthritis is the main cause of disability and pain. Due to limited understanding of the disease mechanism, it is generally difficult to diagnose early and eliminate sequelae. The aim of this study was to identify novel biomarkers for osteoarthritis (OA) using a bioinformatics approach. Method: The gene expression dataset GSE82107 were obtained from the Gene Expression Omnibus (GEO) database. Matrix quality assessment was performed using corrplot packages and principal component analysis. The differentially expressed genes (DEGs) picked out using GEO2R tool. Enrichment analyses were performed using The Database for Annotation, Visualization and Integrated Discovery and Gene Set Enrichment Analysis (GSEA). Weighted correlation network analysis (WGCNA) was used to find gene modules highly associated with OA. Cytoscape with Molecular Complex Detection (MCODE) plug-in was utilized to analyze protein-protein interaction of these DEGs. Receiver operator characteristic curve analysis was used to evaluate the diagnostic effectiveness of genes. Results: Samples with different conditions (HC and OA) are obviously distinguished, and the distance between biological replicates is relatively close. In total, 1676 DEGs were identified. Enrichment analysis showed that there were some gene sets related to OA pathology, such as chondrocyte development. The results of WGCNA analysis showed that 298 genes were most positive associated with OA. 10 common genes obtained were selected as candidate core genes. ROC results showed that 5 of these genes had the greatest diagnostic value. Conclusion: This is the first study to identify biomarkers related to OA by combining multiple algorithms such as GSEA, WGCNA, MCODE and ROC. We suggest that GLG1, PAPSS2, CTSK, TIMP1 and SDC1 could serve as valuable biomarkers. Further studies are needed to examine the precise role and mechanism of these genes in OA.

目的:探讨左归丸治疗绝经后骨质疏松症的分子机制之一是通过对β2-肾上腺素能受体(β2AR)介导的核转录因子-κB受体激活因子配体(RANKL)/骨保护素(OPG)信号通路的调节.方法:通过去卵巢法建立骨质疏松模型,将40只雌性SPF级SD大鼠随机分为模型组、假手术组、雌二醇(50 μg·kg-1·d-1)组、左归丸低、高剂量(2.3,4.6 g·kg-1)组.采用酶联免疫吸附实验检测血清骨转换标志物;显微CT对右侧股骨远端骨密度水平和骨小梁微结构进行测量和分析;实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测胫骨和下丘脑组织中β2AR,OPG,RANK mRNA和蛋白表达水平.结果:与模型组比较,左归丸可显著降低大鼠血清中Ⅰ型胶原交联羧基端肽(β-CTX)水平(P＜0.01),增加骨特异性碱性磷酸酶(BALP)水平(P＜0.01),减少卵巢摘除诱导引起的大鼠骨量丢失,改善股骨远端骨小梁显微结构水平.同时,左归丸高、低剂量组均能显著上调胫骨组织中OPG mRNA和蛋白表达(P＜0.01),下调下丘脑组织β2AR mRNA和蛋白表达水平(P＜0.01),同时也能显著下调胫骨组织中RANKL mRNA和蛋白表达水平(P＜0.01).结论:左归丸改善围绝经期综合症,增加骨量和改善骨小梁显微结构水平的分子机制可能与对β2 AR介导的RANKL/OPG信号通路的调节有关.