Attenuation of diabetic nephropathy by Chaihuang-Yishen granule through anti-inflammatory mechanism in streptozotocin-induced rat model of diabetics

Ethnopharmacological relevance

Traditional Chinese medical herbs have been used in China for a long time to treat different diseases. Based on traditional Chinese medicine (TCM) principle, Chaihuang-Yishen granule (CHYS) was developed and has been employed clinically to treat chronic kidney disease including diabetic nephropathy (DN). The present study was designed to investigate its mechanism of action in treatment of DN.

Materials and methods

Diabetic rats were established by having a right uninephrectomy plus a single intraperitoneal injection of STZ. Rats were divided into four groups of sham, diabetes, diabetes with CHYS and diabetes with fosinopril. CHYS and fosinopril were given to rats by gavage for 20 weeks. Samples from blood, urine and kidney were collected for biochemical, histological, immunohistochemical and molecular analyses.

Results

Rats treated with CHYS showed reduced 24 h urinary protein excretion, decreased serum TC and TG levels, but CHYS treatment did not affect blood glucose level. Glomerular mesangial expansion and tubulointerstitial fibrosis in diabetic rats were significantly alleviated by CHYS treatment. Moreover, CHYS administration markedly reduced mRNA levels of NF-κB p65 and TGF-β1, as well as decreased protein levels of NF-κB p65, MCP-1, TNF-α and TGF-β1 in the kidney of diabetic rats.

Conclusions

CHYS ameliorates renal injury in diabetic rats through reduction of inflammatory cytokines and their intracellular signaling.     

Evaluation of Tableting and Tablet Properties of Kollidon SR: The Influence of Moisture and Mixtures with Theophylline Monohydrate

The aim of the study was firstly to investigate the influence of moisture on the tableting and tablet properties of Kollidon SR and secondly to investigate the influence of theophylline monohydrate on the tableting behavior and tablet properties produced from binary mixtures with Kollidon SR. In comparison to Kollidon SR, microcrystalline cellulose (MCC) was used. The glass transition temperature (T_g) of the powder over the whole range of RH (0–90%), and in addition, the T_g of tablets of Kollidon SR were measured. Densities and flowability of the powders were analyzed. The tablets were produced at five different maximum relative densities (ρ_{rel, max}) on an instrumented eccentric tableting machine. They were produced at three different relative humidities (RH), 30%, 45%, and 60% RH for the pure substances and binary mixtures with different ratios of drug and excipient were tableted at 45% RH. The tableting properties were analyzed by 3D modeling, force-displacement profiles, and compactibility plots. First, the T_g of the powder decreased with increasing RH and the T_g of the tablet was 4–8 K lower than the powder. The predominant deformation of Kollidon SR is plastic deformation and Kollidon SR showed a higher compactibility than MCC. The parameters of the 3D model showed an extreme change between 45 and 60% RH, and at higher RH more and more particles deformed elastically. This was confirmed by analysis of force-displacement profiles. At 60% RH, the radial tensile strength of the Kollidon SR tablets was half of the radial tensile strength at 45% RH. The reason is a higher relative energy of plastic deformation than for MCC. This results in a better utilization of the energy to deform the powder into a tablet and the exceeding of the glass transition temperature at higher RH. In conclusion, at 60% RH at the same ρ_{rel, max}, tableting and tablet properties of Kollidon SR are extremely changed since plasticity is significantly higher. In the second part of the study, the insufficient flowability of theophylline monohydrate can be compensated by using Kollidon SR in a mixture with up to 20% theophylline. Further, pressure plasticity ε of MCC and Kollidon SR was lowered in the mixture with theophylline monohydrate. The same is valid for the compactibility. The influence of theophylline monohydrate on the pressure plasticity e of the mixtures was better compensated in the mixture with MCC than in a mixture with Kollidon SR. This compensation was also visible by analyzing the force-displacement-profiles. However, hardly any influence on the radial tensile strength could be detected. Kollidon SR and Kollidon SR mixtures exhibited a higher compactibility than MCC and MCC mixtures. The differences became smaller with increasing theophylline content.     

用差示扫描量热法及激光拉曼光谱研究足叶乙甙对二棕榈酰磷脂酰胆碱脂质体膜流动性影响

本文用DSC和激光拉曼光谱研究抗癌药物足叶乙甙(4-去甲基表鬼臼毒素-β-D-乙叉吡喃葡萄糖甙,简称VP 16-213)与二棕榈酰磷脂酰胆碱(DPPC)脂质体的作用。VP 16-213分子掺入DPPC脂质体双层中,不但使相转变温度向高温移动,而且吸热峰的半高宽度随VP 16-213浓度增加而变宽。其Raman光谱在频率2850 cm~(-1)处的C-H键对称伸缩振动亦随着药物浓度增加而减弱。这些结果表明VP 16-213分子是定域在脂双层中DPPC分子链的C_1～C_9亚甲基区域,使脂质体的有序性提高而流动性降低。     

Medical visits of childhood cancer survivors in Japan: A cross‐sectional survey

Background: Although more children with cancer continue to be cured, these survivors experience various late effects. Details of the medical visit behaviors of childhood cancer survivors (CCS) in adulthood remain to be elucidated. Methods: In order to examine medical visits in the past and future of CCS, we performed a cross‐sectional survey with self‐rating questionnaires on medical visits of CCS compared with control groups (their siblings and the general population). Results: Questionnaires were completed by 185 CCS, 72 of their siblings and 1000 subjects from the general population and the results were analyzed. Mean ages at this survey and the duration after therapy completions of CCS were 23 and 12 years, respectively. We found that the previous treatment hospitals (where CCS were treated for their cancer) were the most commonly visited medical facilities for the CCS group (74% for female patients and 64% for male patients) and more than half of the CCS preferred to continue visiting the previous treatment hospital with enough satisfaction in Japan. The multivariate analysis showed that female sex and relapse were significantly associated with the past visits to the previous treatment hospital and that the CCS with brain tumors or bone/soft tissue sarcomas and CCS with any late effects tended to continue the relationships with the hospital. In addition female sex was also significantly associated with desired future visits to the previous treatment hospital. On the other hand, the married CCS tended to be disinclined to visit the hospital it in the future. Conclusions: In order to optimize risk‐based care and promote health for CCS after adulthood, we should discuss the medical transition with CCS and their parents.     

内皮-间质转化在眼部疾病中的研究进展

内皮-间质转化(endothelial-to-mesenchymal transition,EndoMT)是在生理或病理条件下,内皮细胞向间质细胞转化或分化,并伴随表型和功能的改变,是纤维修复的重要组成部分,广泛参与胚胎发育、肿瘤侵袭及多种纤维化疾病发展的病理生理过程,有关EndoMT在眼部疾病中作用的研究也取得一定进...     

Chrysophanol localizes in mitochondria to promote cell death through upregulation of mitochondrial cyclophilin D in HepG2 cells

Objective:Chrysophanol(Chry) displays potent anticancer activity in human cancer cells and animal models,but the cellular targets of Chry have not been fully defined.Herein,we speculated whether mitochondria were a target involved in Chry-induced cytotoxicity.Methods:Human liver cancer cell line HepG2 was incubated.The cytotoxicity was evaluated by MTT assay.Mitochondria localization was evaluated by a confocal microscopy.Mitochondrial membrane potential ΔΨm was detected by TMRE staining and determined by the flow cytometer.The levels of ATP,mitochondrial superoxide anions,and GSH/GSSG were determined according to the assay kits.The apoptosis were evaluated through Hoechst33342/PI and Annexin V/PI staining,respectively.The expression of cyclophilin D(CyPD) was determined by immunoblot method,and the interaction between CyPD and Chry was analyzed by molecule docking procedure.Results:Chry itself mainly localized in mitocho ndria to cause mitochondrial dysfunction and cell death in HepG2 cells.As regard to the mechanism,cyclosporin A as the inhibitor for the formation of mitochondrial permeability transition pore(mPTP) moderately suppressed cell death,indicating mPTP involved in the process of cell death.Further,Chry enhanced the protein expression of Cyclophilin D(CyPD) which is a molecular componentry and a modulator of mPTP,while antioxidant N-acetyl-L-cysteine inhibited the expression of CyPD.Molecule docking procedure disclosed two hydrogen-bonds existed in CyPD-Chry complex with-11.94 kal/mol of the binding affinity value.Besides,the mtDNA-deficient HepG_2-ρ0 cells were much resistant to Chry-induced cell death,indicating mtDNA at least partly participated in cell death.A combination of Chry and VP-16 produced the synergism effect toward cell viability andΔΨm,while Chry combined with Cis-Pt elicited the antagonism effect.Conclusion:Taken together,enrichment in mitochondria and actions on mPTP,CyPD and mtDNA provides an insight into the anticancer mechanism of Chry.The combination therapy for Chry with clinical drugs may deserve to further explore.     

基于IRS2/PI3K/FOXO4信号通路研究复方仙草颗粒抑制糖尿病肾病足细胞上皮-间充质转化的作用机制

目的 基于胰岛素受体底物2（insulin receptor substrate 2,IRS2）/磷脂酰肌醇-3-激酶（phosphatidylinositol-3-kinase,PI3K）/叉头框蛋白O4（forkhead box O4,FOXO4）信号通路探讨复方仙草颗粒抑制糖尿病肾病（diabetic nephropathy,DN）大鼠肾足细胞上皮-间充质转化（epithelial-mesenchymal transition,EMT）的作用机制。方法 采用单侧肾切除、高糖高脂饲料喂养及一次性ip链脲佐菌素制备DN模型大鼠,另设置对照组和假手术组,将造模成功的大鼠随机分为模型组、厄贝沙坦（15.75 mg/kg）组和复方仙草颗粒组低、中、高剂量（315.0、472.5、945.0 mg/kg）组,各给药组ig相应药物,连续6周。检测各组大鼠空腹血糖、肾脏指数（kidney index,KI）、尿微量白蛋白/肌酐比值（urinary albumin-to-creatinine ratio,UACR）、尿素氮（blood urea nitrogen,BUN）水平;采用苏木素-伊红（HE）染色法观察肾组织病理变化;采用透射电镜（TEM）观察肾足细胞超微结构变化;采用qRT-PCR和Western blotting检测大鼠肾组织足细胞裂孔膜蛋白（nephrin）、P-钙黏蛋白（P-cadherin）、α-平滑肌肌动蛋白（α-smooth muscle actin,α-SMA）、Desmin、成纤维细胞特异蛋白-1（fibroblast specific protein-1,FSP-1）及IRS2/PI3K/FOXO4信号通路相关因子mRNA和蛋白表达。结果 与对照组比较,模型组大鼠KI、UACR、BUN水平均显著升高（P<0.05）,肾小球肥大,肾小管广泛扩张,上皮细胞变性脱落,足突融合或丢失;肾组织nephrin、P-cadherin表达显著降低（P<0.05）,α-SMA、Desmin、FSP-1表达显著升高（P<0.05）,IRS2/PI3K/FOXO4信号通路相关因子mRNA和蛋白表达均显著降低（P<0.05）。与模型组比较,复方仙草颗粒组大鼠KI、UACR、BUN水平显著降低（P<0.05）,肾脏病理结构和足细胞超微结构明显改善;肾组织nephrin、P-cadherin表达显著升高（P<0.05）,α-SMA、Desmin、FSP-1表达显著降低（P<0.05）,IRS2/PI3K/FOXO4信号通路相关因子mRNA和蛋白表达均显著升高（P<0.05）。结论 复方仙草颗粒能够通过调节IRS2/PI3K/FOXO4信号通路改善DN大鼠肾功能,抑制DN大鼠足细胞EMT,减轻足细胞损伤。     

Cx26和Cx43在乳腺癌中的表达及与上皮间质转化的关系

目的研究连接蛋白26(Connexin26,Cx26)和连接蛋白43(Connexin43,Cx43)在乳腺癌中的表达及与上皮间质转化(EMT)的关系。方法应用免疫组织化学方法检测70例浸润性导管癌、10例乳腺良性病变(乳腺纤维腺瘤)患者中Cx26和Cx43的表达情况,并分析其与EMT的关系。结果Cx26和Cx43在浸润性导管癌中表达的阳性率分别为31%(22/70)和47%(33/70),与乳腺纤维腺瘤中相比,Cx26和Cx43在浸润性导管癌中的表达明显降低(P0.05);在浸润性导管癌中Cx26和Cx43的表达与EMT状态呈负相关(r=-0.26,r=-0.24)。结论Cx26和Cx43与EMT状态呈负相关,可作为乳腺浸润性导管癌中EMT发生的有价值的标记物。     

miR-194-3p靶向调控MMP-11抑制骨肉瘤细胞的增殖、迁移和侵袭

目的:研究miR-194-3p对骨肉瘤细胞增殖、迁移和侵袭的影响,并探讨其机制。方法:实验设置空白对照组（control）、miR-194-3p-mimics组（转染 miR-194-3p-mimics）、miR-194-3p-inhibitor组（转染 miR-194-3p-inhibitor）、siRNA-MMP-11组（转染 siRNA-MMP-11）、ovRNA-MMP-11组（转染ovRNA-MMP-11）、miR-194-3p-mimics+ovRNA-MMP-11组（转染miR-194-3p-mimics+ovRNA-MMP-11）,均以脂质体法转染至人成骨肉瘤细胞Saos-2;MTT法检测细胞增殖率;Transwell实验检测细胞侵袭能力;细胞划痕实验检测细胞迁移能力;蛋白印迹实验（Western blot）检测E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、基质金属蛋白酶 11（MMP-11）蛋白表达;实时荧光定量PCR（qRT-PCR）检测miR-194-3p的表达水平。结果:MTT法检测结果显示:miR-194-3p-mimics组、siRNA-MMP-11组、miR-194-3p-mimics+ovRNA-MMP-11组会降低细胞增殖,并与空白对照组和ovRNA-MMP-11组相比有统计学差异（P＜0.05）。细胞划痕实验检测结果显示:miR-194-3p-mimics组、siRNA-MMP-11组、miR-194-3p-mimics+ovRNA-MMP-11组细胞愈合率明显低于ovRNA-MMP-11组（P＜0.05）。细胞侵袭实验检测结果显示:siRNA-MMP-11组、miR-194-3p-mimics+ovRNA-MMP-11组降低细胞侵袭能力,并与空白对照组和ovRNA-MMP-11组相比有统计学差异（P＜0.05）。qRT-PCR检测结果显示:miR-194-3p-mimics组中miR-194-3p的表达水平与空白对照组、miR-194-3p-inhibitor组、siRNA-MMP-11组、ovRNA-MMP-11组、miR-194-3p-mimics+ovRNA-MMP-11组相比均有统计学差异（P＜0.05）。结论:miR-194-3p在人成骨肉瘤细胞中高表达,并可抑制骨肉瘤细胞的增殖、迁移和侵袭,其机制可能与靶向调控MMP-11所参与的上皮间充质转化（epithelial mesenchymal transition,EMT）信号通道有关,miR-194-3p将来可能为骨肉瘤的预防和治疗提供新靶点。     

TGF-β1诱导的肝癌细胞SMMC-7721上皮间质转化对细胞Wnt/β-catenin表达的影响

目的探讨TGF-β1诱导的肝癌细胞SMMC-7721上皮间质转化对细胞信号通路蛋白Wnt/β-catenin表达的影响。方法采用5μg·mL-1 TGF-β1诱导SMMC-7721发生上皮间质转化,分别采用RT-PCR和Western blot法检测细胞中Wnt1、β-catenin、Vimentin mRNA和总蛋白的表达。结果 TGF-β1能显著诱导肝癌细胞SMMC-7721发生上皮间质转化,细胞中Wnt1、β-catenin、Vimentin mRNA和相应总蛋白的表达均显著增加(P<0.05)。结论 TGF-β1诱导肝癌细胞SMMC-7721上皮间质转化时可促进细胞Wnt/β-catenin的表达。