Buyang Huanwu decoction up-regulates Notch1 gene expression in injured spinal cord

Expression of genes in the Notch signaling pathway is altered in the injured spinal cord, which indicates that Notch participates in repair after spinal cord injury. Buyang Huanwu decoction, a traditional Chinese herbal preparation, can promote the growth of nerve cells and nerve fibers; however, it is unclear whether Buyang Huanwu decoction affects the Notch signaling pathway in injured spinal cord. In this study, a rat model was established by injuring the T10 spinal cord. At 2 days after injury, rats were intragastrically administered 2 m L of 0.8 g/m L Buyang Huanwu decoction daily until sacrifice. Real-time reverse transcription polymerase chain reaction analysis demonstrated that at 7, 14 and 28 days after injury, the expression of Notch1 was increased in the Buyang Huanwu decoction group compared with controls. These findings confirm that Buyang Huanwu decoction can promote the expression of Notch1 in rats with incomplete spinal cord injury, and may indicate a mechanism to promote the repair of spinal cord injury.

Xuezhikang(血脂康) Reduced Renal Cell Apoptosis in Streptozocin-Induced Diabetic Rats through Regulation of Bcl-2 Family

Objective： To investigate the effect of Xuezhikang（血脂康, XZK） on renal cell apoptosis in diabetic rats and the possible mechanism. Methods： Sixty-six rats were randomly divided into 3 groups： the normal, model and XZK groups. In each group, the rats were further randomly divided into 3-month and 6-month subgroups, respectively. Diabetes of rats was induced by a single intraperitoneal injection of 1% streptozocin at 60 mg/kg body weight. Rats in the XZK group received gastric perfusion of XZK（1200 mg/kg body weight） everyday for 3 or 6 months, while rats in the normal and model groups received equal volume of saline. Twenty-four hours＇ urine was collected for urinary albumin excretion（UAE） measurement. Periodic acid-Schiff（PAS） and Masson＇s trichrome staining were used for saccharides and collagen detection. Cell apoptosis of renal cortex was investigated by Td T-mediated d UTP nick end labeling（TUNEL） staining. Bax and Bcl-2 expressions were detected by immunohistochemistry and Western blot, respectively. Cytochrome C（Cyt C） and caspase-9 concentration were detected by Western blot. Results： Compared with the model group, XZK treatment could significantly decrease the kidney hypertrophy index, 24 h UAE, renal cell apoptosis, cytoplasmic Cyt C level and active caspase-9 level, as well as suppress the increment of Bax and up-regulate the expression of Bcl-2, leading to the suppression of Bax/Bcl-2 ratio at 3 and 6 months（P〈0.05 or P〈0.01）. Moreover, XZK treatment could alleviate the deposition of PAS-stained saccharides and Masson＇s trichromestained collagen to different extent. Conclusions： Renal cell apoptosis was observed in diabetic kidney, in which mitochondrial apoptotic pathway might be involved. XZK treatment could attenuate pathological changes in diabetic kidney and reduce renal cell apoptosis, probably via the suppression of Bax/Bcl-2 ratio, which lead to inhibition of Cyt C release and following caspase-9 activation.

Potent Therapeutic Effects of Shouwu Jiangqi Decoction(首乌僵芪汤)on Polycystic Ovary Syndrome with Insulin Resistance in Rats

Objective：To investigate the effect of Shouwu Jiangqi Decoction（首乌僵芪汤,SJD） on polycystic ovary syndrome（PCOS） with insulin resistance（IR） in rats and to explore the underlining molecular mechanisms.Methods：A total of 51 female Sprague-Dawley rats were randomly divided into 6 groups：control group（n=7）,model group（n=8）,SJD high-dose group（n=9）,SJD medium-dose group（n=9）,SJD low-dose group（n=9） and DMBG group（n=9）.Radioimmunoassay was used to measure serum follicle-stimulating hormone（FSH）,luteinizing hormone（LH） and testosterone concentrations and qRT-PCR and western blot were used to examine the expression levels of mRNA and protein respectively of insulin receptor substrate 1（IRS-1）and phosphatidylinositide 3-kinases（PI3K） p85α in different groups.Results：FSH level significantly decreased in the model group compared with the normal control（P〈0.01）,and high-dose SJD and DMBG can significantly increase FSH level（P〈0.01）.LH level showed a mild increase without statistic significance in the model group compared with the control and different dosages of SJD had no significance effect on LH level,while DMBG can significantly decrease LH level（P〈0.01）.Testosterone level significantly increased in the model group compared with the control group（P〈0.01）,and high-dose SJD and DMBG can significantly decrease testosterone level（P〈0.01）.The expression of IRS-1 as well as PI3Kp85α were significantly decreased in the model group compared with the normal control group at both mRNA（P〈0.001） and protein（P〈0.01） level,and both high-dose SJD and DMBG can enhance IRS-1 and PI3 K expression（P〈0.05）.Conclusions：SJD has potent therapeutic effects on PCOS with IR in rats.The therapeutic effects of SJD on IR and ovulatory dysfunction are probably achieved through correcting the defective insulin signaling transduction.

Differentially Expressed Genes in Heads and Tails of Angelica Sinensis diels： Focusing on Ferulic Acid Metabolism

Objective： To explore the scientific connotation of the discrepant pharmaceutical activities between the head and tail of Angelica sinensis diels（AS）, an important herb extensively utilized in Chinese medicine, by the approach of transcriptome sequencing. Methods： Ten samples of AS were randomly collected in Min County, Gansu Province of China. Transcriptome sequencing of AS was accomplished in a commercial ILLumina Hi Seq-2000 platform. The transcriptome of each head and tail of AS were fixed in a gene chip, and detected under the procedure of Illumina Hi Seq-2000. Differentially expressed unigenes between the heads and tails of AS were selected by Shanghai Biotechnology Corporation（SBC） online analysis system, based on Gene Ontology, Kyoto Encyclopedia of Genes and Genomes and relevant bioinformatic database. Results： Totally 63,585 unigenes were obtained from AS by high-throughput sequencing platform. Among which 3359 unigenes were identified as differentially expressed unigenes between the heads and tails of AS by SBC analysis system scanning. Of which 15 differentially expressed unigenes participate in the metabolic regulation of phenylpropanoid biosynthesis（PB） pathway and ferulic acid metabolites, in response to the distinguished pharmaceutical actions of the heads and tails of AS. Conclusion： Different content of ferulic acid in the heads and tails of AS is related to the differentially expressed genes, particularly involved in the PB pathway.

Inhibition Mechanism of Qingluo Tongbi Granule(清络通痹颗粒)on Osteoclast Differentiation Induced by Synovial Fibroblast and Monocytes Co-Culture in Adjuvant-Induced Arthritic Rats

Objective： To study the mechanism underlying the inhibitory effect of Qingluo Tongbi Granule （清络通痹颗粒, QTG） on osteoclast differentiation in rheumatoid arthritis in rats. Methods： Fibroblast and monocyte co-culture were used to induce osteoclast differentiation in adjuvant-induced arthritic （AIA） rats. Serum containing QTG was prepared and added to the osteoclasts, and activation of the tumor necrosis factor receptorassociated factor 6/mitogen-activated protein kinase/nuclear factor of activated T cells, cytoplasmic1 （TRAF6/ MAPK/NFATcl） pathways was examined. Results： The induced osteoclasts were multinucleated and stained positive for tartrate-resistant acid phosphatase （TRAP） staining. Serum containing QTG at 14.4, 7.2 or 3.6 g/kg inhibited the activation of TRAF6, extracellular regulated protein kinase （ERK）1/2, c-Jun N-terminal kinase （JNK） and p38 and decreased the percentage of cells with nuclear NFATcl in a dose-dependent manner, the high and middle doses exhibited clear inhibitory activity （P〈0.01 and P〈0.05, respectively）. After the addition of MAPK inhibitors, the NFATcl expression showed no significant difference compared with the control group （P〉0.05）. Conclusions： Serum containing QTG could generally inhibit the TRAF6/MAPK pathways and possibly inhibit the NFATcl pathway. In addition, QTG may regulate other signaling pathways that are related to osteoclast differentiation and maturation.