This monthly, peer-reviewed journal includes studies devoted to molecular medicine, underscoring aspects including pharmacology, pathology, genetics, neurosciences, infectious diseases, molecular cardiology and molecular surgery. In vitro and in vivo studies of experimental model systems pertaining to the mechanisms of a variety of diseases offer researchers the necessary tools and knowledge with which to aid the diagnosis and treatment of human diseases.

目的 观察蒲葵子乙醇提取物(EELC)对肝癌细胞以及裸鼠皮下移植瘤的抑制作用,探讨EELC阻滞细胞周期的抗肝癌作用机制.方法 培养人肝癌HepG2细胞,采用MTT法和平板克隆形成实验检测不同浓度EELC(0、0.125、0.25、0.5 mg/mL)对HepG2细胞活力和细胞周期的影响.构建裸鼠HepG2皮下移植瘤模型,将裸鼠随机分为对照组和EELC组,分别给予生理盐水和EELC灌胃液[3 g/(kg·d)]灌胃给药,连续21 d后,剥取肿瘤组织,采用免疫组织化学方法分析肿瘤组织中PCNA、CDK1、CyclinB1和p21的蛋白表达.结果 细胞实验结果显示,与0 mg/mL组比较,0.125、0.25、0.5 mg/mL组HepG2细胞活力和克隆形成率均明显下降(P<0.05),呈剂量依赖;与0 mg/mL组比较,0.125、0.25、0.5 mg/mL组细胞周期进程明显抑制,阻滞于G2/M期.动物实验结果显示,与对照组比较,EELC组小鼠肝癌组织PCNA、CDK1和CyclinB1蛋白表达明显降低(P<0.05),p21表达明显提高(P<0.05).结论 EELC通过促进p21表达,抑制PCNA、CDK1和CyclinB1表达,使细胞周期阻滞于G2/M期,抑制肝癌细胞增殖,进而抑制肝癌生长.

Doxorubicin (Dox), an effective antineoplastic drug, was limited use for cardiotoxicity. Xinshuitong Capsule (XST), a patented herbal formula, showed desirable beneficial effects in the treatment of chronic heart failure (CHF) patients. However, the drug on Dox-induced cardiotoxicity remains unclear. Ninety male Sprague-Dawley rats were randomized into two groups: 15 rats were selected as the normal group and 75 rats were injected intraperitoneally with Dox to establish CHF rat models, the success ones were randomly divided into five groups: low XST (LXST), medium XST (MXST) or high XST (HXST) (4.9, 9.8, or 19.6 g/kg d) administrated intragastrically twice a day for 4 weeks, with the captopril-treated group and the model group as comparison. The model group showed the cardiac functions generally impaired, and CHF mortality rate higher (47%) than those in the XST-treated groups (averaged 24%, P < 0.05). Compared with XST-treated groups, myocardial remodeling, inflammation and desarcomerization, and higher water content more severe in the cardiac tissue in the model group (P < 0.05), which was associated with higher expressions of mRNA or protein levels of AQP1, 4 and 7. Dox-impaired cardiac functions, cardiac remodeling and myocardial edema could be dose-dependently reverted by XST treatment. XST could inhibit AQP1, 4 and 7 at mRNA levels or at protein levels, which was associated with the attenuation of myocardial edema and cardiac remodeling, decreasing the ventricular stiffness and improving the cardiac functions and rats' survival. AQPs is involved in cardiac edema composed one of the mechanisms of Dox-induced cardiotoxicity, XSTvia inhibition of AQPs relieved the Dox-induced side effects.

Background Liver metastasis is a common cause of death from colorectal cancer (CRC). In this paper we developed a liver metastasis mouse model by microsurgical orthotopic implantation (MSOI) to illuminate the CRC progression with an eye toward developing effective drug treatment. Methods Murine colon carcinoma CT-26 cells were cultured and then injected to male BALB/c athymic nude mice right flank to generate subcutaneous implantation tumor with 2×107 CT-26 cell suspension in DMEM. Tumor tissue at an average size of 1 cm3 was injected into another nude mice right flank with 20-gauge inoculating needle. Between fourth and sixth generations, tumor tissue sewn into the cecal surface establishes orthotopic transplanted CRC model by MSOI. Then on the 7th, 14th, 21st and 35th day, body weight, abdomen circumference, volume of ascites and local tumor weight were observed and weighed. On the 21st day and 35th day, local tumor rate was calculated, and metastatic tumors of other organs were observed. Tumor tissue was stained by HE for pathologic analysis. Results On the 35th day, body weight and abdomen circumference of the model group were significantly higher than the control group (P<0.01). Local tumor weight increased rapidly from the 21st d to the 35th d (P<0.01), and take rate was high (100%). Metastatic tumor appeared only in liver on the 21st day and then invaded to liver, stomach, retroperitoneal lymph node and abdominal wall on the 35th day. The metastatic rate of liver tumor respectively was 83.3% and 100% on the 21st day and 35th day, but liver function remained normal. Pathologic analysis showed that colorectal tumor invaded the normal tissue of liver, abdominal wall and stomach. Conclusions A stable hepatic metastasis mouse model of murine CRC was established by MSOI.

Objective: This study aimed to explore the neuroprotective effect of Baihe Dihuang Tang (BDT), a traditional Chinese herbal decoction used for nervous-mental system diseases, on serum-deprived PC12 cell. Methods: BDT treatment time and concentration were determined through 4,5-dimethylthiazol-2-y1,2,5-diphenyl tetrazolium (MU) assay. PC12 cells were randomly divided into three groups: control (cells cultured in serum-containing medium), model (cells cultured in serum-deprived medium), and BDT (cells cultured in serum-deprived medium and treated with 3 mg/mL BDT for 24 h) groups. Cell morphology was observed under an inverted phase contrast microscope. The ultrastructure of cells was studied under a transmission electron microscopy. Membrane potential was determined using a laser scanning confocal microscope. Total protein and ATP levels were analyzed by the bicinchoninic acid method and firefly luciferase method, respectively. Results: Compared with the model group, PC12 cells in the BDT group exhibited extensive morphology and adhered to the culture plate. The ultrastructure was modified and possessed a smooth membrane and nuclear surface, increased microvilli-like membrane protrusions, homogeneously distributed organelles, rare pinocytosis and phagocytosis, abundant ribosomes, rare vacuoles, and normal-shaped nucleus. BDT also enhanced the total cell protein content, ATP level, and membrane hyperpolarization. Conclusion: BDT could reduce cell death and neural excitability and regulate energy metabolism caused by serum deprivation. We suggest that BDT could modify the hypometabolic state and neural overactivity in chronic fatigue syndrome (CFS). This work may provide new insights into the possibility of using BDT as a therapeutic agent for CFS.

Objective:Up to now,morbidity and mortality of hepatocellular carcinoma (HCC) have been increasing all over the world especially in China.5-fluorouracil (5-FU) is a common used chemotherapy drug for many types tumor in clinic including HCC.However with the bigger dosage of 5-FU applying the more severely toxicity and side-effects have limited it's widely clinical use.Therefore,there is a strong demand to find some new drugs of low side-effect for HCC treatment.Accumulating attention has been paid to Traditional Chinese Medicine (TCM) because of the multi-targets,low-toxicity and the whole system regulation for patients.Maybe it is a potential effective way on finding some TCM to enhance anti-tumor effect with low dose 5-FU.Jiedu Xiaozheng Decoction (JXD),a polyherbal formula of TCM,has been used in clinical treatment for digestive tract tumor especially HCC for more than a decade.The previous studies found that JXD inhibited tumor growth by inducing apoptosis,blocking cell cycle and anti-angiogenesis.In this study,we evaluated anti-cancer effects of JXD ethyl acetate extract (EE-JXD) combined with 5-FU in vitro and in vivo in order to explore the biological mechanism and provide more experimental basis for clinical application of JXD.Methods:EE-JXD was prepared according to the describe in the previous study.Then EE-JXD was diluted to 200 mg/mL with dimethyl sulfoxide for in vitro experiment and was dissolved in normal saline to a final concentration of 6 mg/mL in vivo study.HepG2 cell line was cultured in RPMI-1640 medium supplemented with 10％ fetal bovine serum,1％ penicillin and streptomycin at 37 ℃ in an incubator containing 5％ CO2.In vitro,cells viability was detected by MTT assay after HepG2 cells were treated with different concentrations of EE-JXD (0.05,0.1,0.2,0.4 mg/mL),5-FU (0.05,0.1,0.2,0.4 mg/mL),and EE-JXD+5-FU (0.05+0.05,0.1+0.1,0.2+0.2,0.4+0.4 mg/mL) for 24 hours.And then the HepG2 cells were divided into control group,EE-JXD group (0.1 mg/mL),5-FU group (0.1 mg/mL) and EE-JXD+5-FU group (0.1mg/mL+0.1 mg/mL).Cells apoptosis of four groups were evaluated by fluorescence assay and flow cytometry respectively after treated for 24 hours.In fluorescence assay,the HepG2 cells were stained with Hoechst 33258 at room temperature and apoptosis was observed under a fluorescence microscope.The treated HepG2 cells were processed and analyzed using an Annexin V-FITC/propidium iodide (PI) assay with flow cytometry for quantita tive determination of early and late stage apoptotic cells.Thirty-two male BALB/c (nu/nu) mice (body weight,18-22 g) were used to establish the subcutaneous xenotransplanted tumor model of HepG2 cell line.HepG2 cells in density of 5×106 cells/mL were subcutaneously injected in the right flank of each mouse.After seven days thirty-two mice were randomly assigned to four groups:EE-JXD group,administered 15 g/(kg ·d) EE-JXD (i.g.);5-FU group,administered 10 mg/(kg· d) 5-FU (i.p.),a low dose confirmed by pre-experiment;EE-JXD+5-FU group,administered 15 g/(kg· d) EE-JXD and 10 mg/(kg· d) 5-FU;and control group (normal saline).Tumor volume and body weight were measured every three days.Tumor weight was weighed after the tumor was stripped at day 30.The apoptosis rate of tumor was evaluated by terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL).Results:EE-JXD,5-FU and EE-JXD+5-FU inhibited proliferation of HepG2 cells in a concentration-dependent manner.The combination of JXD and 5-FU had a strong inhibitory effect on the proliferation of HepG2 (P＜0.01,compared with the control group).Apoptosis was obvious in both 5-FU and EE-JXD groups in fluorescence assay.Apoptosis were enhanced when EE-JXD combined with 5-FU.Further assay using Annexin V-FITC/PI double staining showed HepG2 cells treated with 5-FU,EE-JXD and EE-JXD+5-FU to induce apoptosis in 5％,7.8％ and 12.9％ after 24 hours.In contrast,mice treated with either 5-FU,EE-JXD or 5-FU plus EE-JXD showed slower progression of established tumors.Control mice reached a total tumor weight of (0.43±0.06) g within 30 days.At this time point,the tumor weight of mice treated with 5-FU was significantly smaller (P＜ 0.01) at (0.08±0.01) g.Significant slowdown in tumor progression compared with control mice was observed in the group of mice receiving 5-FU and EE-JXD,reaching only (0.05±0.01) g tumor volume with in 30 days (P＜ 0.01).During 30 days' treatment,the body weight between the control group and the treatment groups were similar (P＞0.05).Apoptosis index was significantly higher in the 5-FU group compared to the control group (P＜ 0.05),and was further increased when EE-JXD combined with 5-FU (P＜ 0.01).Conclusion:The present study shows that low-dose 5-FU combined with EE-JXD has a strong anti-cancer effect than 5-FU alone.The effect is more obvious in vivo than in vitro experiments.No adverse events appeared in mice treated with both 5-FU and EE-JXD,indicating that 5-FU combine with EE-JXD at the indicated dose is safe.The results suggest that EE-JXD may potentially be combined with 5-FU for HCC and warrants additional investigation.

目的:分析透骨消痛胶囊对骨质疏松性骨关节炎模型骨重建与炎症因子的影响,为治疗骨质疏松性骨关节炎提供依据.方法:建立骨质疏松性骨关节炎大白兔模型,取模型兔与正常对照兔各6只比较,鉴定造模成功.再将正常对照组与模型对照组大白兔各12只用生理盐水灌胃,中药组大白兔12只用透骨消痛胶囊灌胃;维持4,8周时,每组各6只取材,检测血清骨重建关键因子、骨形成与骨吸收标志物以及股骨髁炎症因子蛋白.结果:中药组与模型对照组比较,碱性磷酸酶、抗酒石酸酸性磷酸酶和骨保护素(OPG)、破骨细胞异化因子(RANKL)活性显著降低,OPG/RANKL比值显著升高(P<0.05或P<0.01);白细胞介素-1β、肿瘤坏死因子-α显著降低(P<0.01或P<0.05).结论:透骨消痛胶囊能下调过度的骨重建速率,调节异常骨代谢水平,降低炎症因子水平,改善骨关节的软骨下骨与软骨变化,延缓骨质疏松性骨关节炎进程.

BACKGROUND:Inflammatory cytokines enhanced the progress of the pathogenesis of osteoarthritis, however the mechanisms remain unclear. The objective is to determine aquaporins (AQPs) in the pathogenesis of osteoarthritis.METHODS AND FINDINGS:Primary rat articular chondrocytes were treated with IL-1β to mimic the early stage of osteoarthritis in vitro. Early osteoarthritis animal model was established by intra-articular injection of 4% papain. Micro- or ultra-structure histopathologic changes, cell viability, apoptosis cells and cell membrane permeability, locations and expressions of AQP1 and AQP3 and matrix were detected in the cartilage or in the chondrocytes of knee. IL-1β could reduce the chondrocytes viability, increase the apoptosis cells, and also impair the cell membrane and organelles. IL-1β significantly induced the up-regulation of AQP1 and AQP3 in the chondrocytes. In the chondrocytes, AQPs were mainly clustered in both membrane and perinuclear region of cytoplasm, while higher AQPs were detected in the superficial and middle layers of the cartilage. With the up-regulation of AQPs, the cartilage matrix was considerably decreased in both the chondrocytes and in the osteoarthritis cartilage. In the early osteoarthritis rat model, serum and synovial fluid confirmed that higher IL-1β could increase the expressions of AQPs, and decrease the cartilage matrix in both the chondrocytes and the cartilage.CONCLUSIONS:Inflammatory cytokine IL-1β via up-regulation of AQPs caused the abnormal metabolism of water transport and loss of the cartilage matrix in the chondrocytes, and ultimately exacerbated the pathogenesis of early osteoarthritis. Therefore, AQPs may be a candidate therapeutic target for prevention and treatment of osteoarthritis.

Previous studies have shown that Tougu Xiaotong capsule (TGXTC) has therapeutic effects on knee osteoarthritis (OA) through multiple targets. However, the mechanisms of action underlying its regulation of subchondral bone reconstruction remain unclear. In this study, we investigated the effects of TGXTC on subchondral bone remodeling. Eighteen six-month-old New Zealand white rabbits of average sex were randomly divided into the normal, model and TGXTC groups. The rabbit knee OA model was induced by a modified Hulth's method in the model and TGXTC groups, but not the normal group. Five weeks postoperatively, intragastric administration of TGXTC was performed for four weeks. After drug administration, the medial femoral condyle and tibia were prepared for observation of cartilage histology via optical microscopy and micro-computed tomography, the serum was collected for biochemical parameters assay and the subchondral bone isolated from the lateral femoral condyle was collected for detection of IL-1β and TNF-α mRNA and protein by reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. The results showed that treatment with TGXTC significantly mitigated cartilage injury and subchondral bone damage, improved the parameter of subchondral trabecular bone, decreased alkaline phosphatase and tartrate-resistant acid phosphatase activity, and significantly reducing the osteoprotegerin/receptor activator of nuclear factor-κB ligand ratio, reduced the expression of IL-1β and TNF-α mRNA and protein. These results suggest that TGXTC could delay the pathological development of OA by regulating subchondral bone remodeling through regulation of bone formation and bone resorption and its relating inflammatory factors, and this may partly explain its clinical efficacy in the treatment of knee OA.

Liver damage results from a variety of insults, including hepatitis and chemical toxicity from alcohol, drugs and other toxins. The present study evaluated the hepatoprotective effects and potential mechanisms of action of the Traditional Chinese Medicine Pien Tze Huang Gan Bao (GB) in a rat model of carbon tetrachloride (CCl4)-induced liver injury. Sixty male Sprague-Dawley rats were randomly divided into six different groups: i) Control, ii) CCl4 injury model and groups treated with iii) silymarin as a positive drug control, iv) 150 mg/kg GB, v) 300 mg/kg GB and vi) 600 mg/kg GB. Control rats received no treatment, while the remaining ones were intraperitoneally injected with CCl4 (2 ml/kg) to induce acute liver disease. Silymarin or GB was orally administered prior to CCl4 treatment in various treatment groups for 7 days. Animals were sacrificed 24 h post-CCl4 injection. It was revealed that GB significantly reduced serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma glutamyl transpeptidase and total bilirubin levels in the serum induced by CCl4. BG also prevented CCl4-induced changes in liver tissues, as revealed by histopathological analysis. CCl4-induced reductions in endogenous liver antioxidant enzyme activities of superoxide dismutase, glutathione and glutathione peroxidase as well as increases in malondialdehyde and thiobarbituric acid reactive substances were inhibited by GB treatment. Activated NF-κB in liver tissues was also significantly increased by CCl4, which was attenuated by GB as indicated by immunohistochemical and PCR analysis. Furthermore, CCl4-mediated increases in the inflammatory factors tumor necrosis factor-alpha and interleukin-1β secretion into the serum and their expression in liver tissues were reversed following GB treatment, as revealed by ELISA and PCR, respectively. These findings suggested that GB protects against CCl4-induced hepatic injury, inflammation and oxidative damage in rats and may be useful in future clinical application of liver injury and disease.

The present study aimed to detect the impact of the ethanol extract of the Livistona chinensis seed (EELC) on angiogenesis in human umbilical vein endothelial cells (HUVECs). A chorioallantoic membrane (CAM) assay was used to detect the anti-angiogenic activity of EELC in vivo. In vitro, the effect of EELC on the proliferation, migration and angiogenesis of HUVECs was determined by an MTT assay, a wound healing assay and a tube formation assay, respectively. The vascular endothelial growth factor (VEGF)-A and VEGF receptor (VEGFR)-2 protein and mRNA level were measured with ELISA and reverse transcription-semi-quantitative polymerase chain reaction. It was observed that EELC significantly decreased the formation of new vessels in the CAM assay. EELC inhibited the proliferation and migration of HUVECs. The extent of tube formation by HUVECs was also reduced by EELC. In addition, EELC treatment reduced the level of VEGF-A and VEGFR-2 mRNA and protein. The results suggest that EELC inhibits tumor angiogenesis through inhibiting the proliferation and migration of HUVECs, and by downregulating VEGF and VEGFR.

目的:观察透骨消痛胶囊对骨质疏松性骨关节炎模型软骨与软骨下骨结构的影响,为探讨治疗该类型骨关节炎提供依据.方法:将18只新西兰大白兔分为正常组、模型组、中药组.除正常组外,其余2组先诱导骨质疏松,再建立膝骨关节炎模型.正常组与模型组给予生理盐水,中药组给予透骨消痛胶囊灌胃4周.HE和番红-固绿染色,光镜观察软骨结构;显微CT观察软骨下骨,分析骨小梁参数;扫描电镜观察软骨下骨微细结构.结果:模型组软骨损伤,基质蛋白多糖浅染,Mankin评分达中期损伤;软骨下骨骨小梁结构破坏,参数变化;胶原纤维等微细结构损伤.中药组软骨损伤减轻,基质深染,Mankin评分为早期损伤;软骨下骨骨小梁结构和参数改善;胶原纤维等微细结构改善.结论:透骨消痛胶囊能改善骨质疏松性骨关节炎模型软骨结构与基质病变,减轻软骨下骨微结构破坏,故可延缓该类型骨关节炎病理进程.

目的 探讨扶正抑瘤方对H22原位肝癌小鼠CD4+ CD25+调节性T淋巴细胞(Tregs)的影响.方法 建立小鼠原位肝癌模型,随机分为模型组(荷瘤小鼠灌胃生理盐水)和药物组(荷瘤小鼠灌胃扶正抑瘤方18 g· kg-1·d-1),另设空白组(正常小鼠灌胃生理盐水),每组8只.连续给药2周后处死,测量肿瘤体积,计算抑瘤率;流式细胞术检测外周血CD3+、CD4+、CD8+T淋巴细胞及CD4+CD25+ Tregs的百分比;Western blot检测瘤组织中又头样转录因子3(Foxp3)和诱导的T细胞共刺激分子(ICOS)的蛋白表达量.结果 药物组的瘤体体积明显小于模型组(P＜0.05),抑瘤率达62.56％;模型组CD3+、CD4+T淋巴细胞含量较空白组明显减低(P＜0.05),CD4+ CD25+ Tregs较空白组明显升高(P＜0.05).药物组给药后CD3+、CD4+T淋巴细胞含量较模型组有明显升高(P＜0.05),CD4+ CD25+ Tregs则明显下降(P＜0.05),Foxp3和ICOS蛋白水平也较模型组明显下降(P＜0.05).结论 扶正抑瘤方可以通过调控CD4+ CD25+ Tregs的数量和功能,提高H22原位肝癌小鼠的细胞免疫功能,发挥抑制肿瘤生长的作用.

Osteoporotic osteoarthritis is a phenotype of osteoarthritis (OA) manifested as fragile and osteoporotic subchondral bone. However, the ultrastructural features of subchondral bone in osteoporosis OA have not been determined. The study was aimed to investigate the ultrastructural dynamic changes of subchondral bone in osteoporotic OA model and how the ultrastructural damage in the subchondral bone caused by osteoporosis deteriorated the cartilage damage in OA. Eighteen rabbits were equally randomized to three groups, including the control, the OA and the osteoporotic OA groups. The structural changes of cartilage were evaluated by HE and safranin-O fast green staining, the Mankin's grading system was used to assess the stage of OA progression. And microstructural or ultrastructural changes in subchondral bone were assessed by micro-computed tomography or by scanning electron microscopy. According to the changes of cartilage histopathology, the OA group was in the early pathological stage of OA while the osteoporotic OA group was in the middle stage of OA based on Mankin's grading system. In addition, the damage of cartilage surface, reduction in the number of chondrocytes and the matrix staining were more increased in the osteoporotic OA group compared to the OA group. Compared to the OA group, the subchondral bone in the microstructure and ultrastructure in the osteoporotic OA group showed more microfracture changes in trabecular bone with more destructions of the tree-like mesh. Moreover, the collagen fibers were random rough with a fewer amount of bone lacunae in subchondral cortical plate in the osteoporotic OA group compared to the OA group. These findings indicated that the subchondral bone ultrastructure in the osteoporotic OA model was characterized by the destruction of the network structure and collagen fibers. The subchondral bone ultrastructural damage caused by osteoporosis may change mechanical properties of the upper cartilage and aggravate OA cartilage. Therefore, early diagnosis and treatment of osteoporosis is of great significance to prevent early OA from further developing osteoporotic OA.

[目的]观察针刺对多囊卵巢综合征胰岛素抵抗(PCOS-IR)模型大鼠生殖内分泌及代谢的影响.[方法]采用来曲唑灌胃联合高脂膳食建立PCOS-IR模型,模型大鼠随机分为模型对照组、针刺组、二甲双胍组,每组6只;另设6只同期喂养的15周龄雌性SD大鼠作为空白对照组.针刺组予针刺穴位(一组为关元、三阴交,另一组为肝俞、脾俞、肾俞穴)治疗,每天1次,两组穴位轮替使用,连续治疗20 d;二甲双胍组给予二甲双胍溶液灌胃;模型对照组给予与针刺组相同的抓取、固定、穴位剪毛、消毒,不做其他任何治疗;空白对照组自由摄食、饮水.各组大鼠治疗后检测血清性激素,观察体重、动情周期和卵巢组织学变化,测定血脂水平、糖耐量、空腹血糖(FPG)和空腹胰岛素(FINS)浓度,并计算胰岛素抵抗指数(HOMA-IR).[结果]①模型对照组大鼠卵巢呈多囊样改变,卵巢闭锁卵泡增多,闭锁卵泡的直径较大,颗粒细胞层数有所减少,未见黄体分布;针刺组和二甲双胍组卵巢恢复排卵,切片可见新鲜黄体分布.②与模型对照组比较,针刺组血清T、LH、PRL水平明显降低,FSH、E2、P水平明显升高(P＜0.05或P＜0.01).③与模型对照组比较,针刺组及二甲双胍组治疗后大鼠体重增加速度明显减慢(P＜0.01),肥胖指数(Lee's指数)明显降低(P＜0.01);血清FINS含量及HOMA-IR显著降低(P＜0.01),糖耐量曲线下面积明显减少(P＜0.05或P＜0.01);血清总胆固醇水平显著降低(P＜0.01),血清低密度脂蛋白显著降低(P＜0.01),针刺组与二甲双胍治疗组差异无统计学意义.[结论]针刺能够改善PCOS-IR模型大鼠生殖内分泌异常和胰岛素抵抗.

目的:观察痰脂消汤对来曲唑灌胃联合高脂膳食诱导的多囊卵巢综合征胰岛素抵抗(PCOS-IR)模型大鼠卵巢胰岛素信号转导途径的影响.方法:来曲唑灌胃联合高脂膳食喂养SD大鼠建立PCOS-IR模型,大鼠随机分为模型组、痰脂消汤组(35 g·kg-1·d-1)、二甲双胍组(200 mg·kg-1·d-1),连续灌胃20 d;另设正常组.实验结束后,腹主动脉采血,测定血清空腹血糖(FPG)和空腹胰岛素(FINS)浓度,并计算胰岛素抵抗指数(HOMA-IR);实时荧光定量PCR(Real-time PCR)检测大鼠卵巢胰岛素受体(INSR),胰岛素受体底物(IRS)-1,酯酰肌醇-3激酶(PI3K),蛋白激酶B(Akt) mRNA表达;蛋白免疫印迹法(Western blot)检测卵巢组织INSR,IRS-1,PI3K,Akt蛋白及其相应磷酸化水平表达.结果:与空白组比较,模型组大鼠空腹胰岛素(FINS),HOMA-IR水平均显著升高(P＜0.01),FPG无明显升高;模型组大鼠卵巢INSR,IRS,PI3K,Akt mRNA表达水平明显降低(P ＜0.05);INSR,p-INSR,IRS,p-IRS,PI3K,p-PI3K,Akt,p-Akt等蛋白表达水平明显降低(P ＜0.05);INSR,IRS,PI3K及Akt磷酸化比例明显降低(P＜0.05).与模型组比较,中药和二甲双胍治疗后大鼠FINS,HOMA-IR均显著降低(P＜0.01);大鼠卵巢INSR,IRS,PI3K,Akt mRNA表达水平均明显升高(P ＜0.05);INSR,p-INSR,IRS,p-IRS,PI3K,p-PI3K,Akt,p-Akt等蛋白表达水平明显升高(P ＜0.05);INSR,IRS,PI3K及Akt磷酸化比例明显升高(P＜0.05).结论:PCOS大鼠存在IR,卵巢内PI3 K/Akt信号途径转导存在异常.痰脂消汤能够显著改善大鼠胰岛素抵抗,可能与调控胰岛素信号转导通路中关键的蛋白表达有关.

Angiogenesis, which has a critical role in human tumor growth and development, is tightly regulated by the Notch signaling pathway. Total alkaloids are active components of the plant Rubus alceifolius Poir, which is used for the treatment of various types of cancer. A previous study by our group showed that the total alkaloids of Rubus alceifolius Poir (TARAP) induced hepatocellular carcinoma (HCC) cell apoptosis through the activation of the mitochondria-dependent pathway in vitro and in vivo, as well as inhibited angiogenesis in a chick embryo chorioallantoic membrane model. In the present study, to further analyze the specific mechanisms underlying the antitumor activity of TARAP, a HCC xenograft mouse model was used to assess the effect of TARAP on angiogenesis in vivo. TARAP was found to suppress the expression of vascular endothelial growth factor (VEGF) A and VEGF receptor-2 in tumor tissues, which resulted in the inhibition of tumor angiogenesis. In addition, TARAP treatment was observed to inhibit the expression of Notch1, delta-like ligand 4 and jagged 1, which are key mediators of the Notch signaling pathway. The present study identified that the inhibition of tumor angiogenesis through the suppression of the Notch signaling pathway may be one of the mechanisms through which TARAP may be effective in the treatment of cancer.

Tumor necrosis factor-α (TNF-α) plays an important role in the abnormal metabolism of osteoblasts (OBs), which leads to subchondral bone (SB) alterations in osteoarthritis. In the present study, Tougu Xiaotong capsule (TXC), a traditional Chinese medicine, was used to treat TNF-α-injured OB-like cells. The cellular viability, mortality and ultramicroscopic morphology were evaluated. Thereafter, the activity of alkaline phosphatase (ALP), secretion of osteocalcin (OCN) and mineralization of nodules were analyzed. The results showed that TXC treatment significantly promoted cell proliferation, reduced cellular mortality and improved cellular ultrastructure, particularly that of the endoplasmic reticulum and nucleus. These data indicate that TXC is able to promote cell growth, as well as prevent inflammation in OB-like cells. Furthermore, the activity of ALP, secretion of OCN and mineralization of nodules were accelerated, and the calcium content of the TNF-α-injured OB-like cells was promoted by TXC treatment. These results indicate that TXC protected the OB-like cells from TNF-α-induced injuries. This may be a potential mechanism through which TXC regulates SB remodeling in the clinical treatment of osteoarthritis.

Metastasis is the leading cause of cancer-related mortality in almost all types of cancers, including colorectal cancer (CRC). Epithelial-mesenchymal transition (EMT) is a critical process during the metastatic cascade. This process may be a potential target for the diagnosis and treatment of CRC. Pien Tze Huang (PZH), a well-known traditional Chinese formula, has been demonstrated to be clinically effective in treating various types of human malignancies, including CRC. Our published data suggest that PZH can induce apoptosis, as well as inhibit cell proliferation and tumor angiogenesis, thus suppressing CRC growth in vitro and in vivo. We evaluated the therapeutic efficacy of PZH against CRC metastasis using a CRC liver metastasis mouse model to further explore the mechanisms underlying the antitumor action of PZH. MTT, migration, and Matrigel invasion assays were used to assess the effect of PZH on cell viability, migration and invasion. We then established an orthotopic liver metastasis model of colon cancer using microsurgical techniques. Mice were intragastrically administered 234 mg/kg/day dose of either PZH or saline for 14 days. The body and tumor weights of the mice were measured after they were sacrificed. Moreover, we examined the effect of PZH inhibition on liver metastasis. Finally, EMT-related proteins and the TGF-β signaling pathway were assessed using immunohistochemical staining (IHS). The present data revealed that PZH significantly inhibited the migration and invasion of CT-26 cells in a dose-dependent manner, which affirmed the inhibitory effect of PZH on CRC cell metastasis. No significant change was observed between the in vivo primary tumor growth and body weight. However, the control group had five cases of liver metastasis (5/6), whereas one case was found in the PZH group (1/6). Thus, PZH exhibited therapeutic efficacy against CRC metastasis without apparent toxicity. The inhibitory effect of PZH on EMT resulted in an increase in E-cadherin expression, as well as a decrease in N-cadherin expression. In addition, PZH significantly inhibited TGF-β, as well as the phosphorylation of Smad2/3 and Smad4 in the tumor tissues, indicating its suppressive action on TGF-β signaling. These molecular effects ultimately resulted in the inhibition of cancer cell EMT and tumor metastasis.