Herbs protect against osteoporosis through anti-inflammation action / 中国组织工程研究

BACKGROUND: The occurrence and development of osteoporosis are shown to be directly related to the inflammatory response induced by immune dysfunction. OBJECTIVE: To summarize the mechanisms of osteoclast differentiation and formation at cellular and molecular levels, as well as the underlying mechanisms of several kinds of medical herbs against osteoporosis, thus paving ways for finding more effective and safe herbs for anti-osteoporosis.METHODS: PubMed database was retrieved using the English keywords of (osteoporosis OR bone loss) AND lipopolysaccharide AND bone resorption", and WanFang database was searched with the Chinese keywords of "osteoarthritis, receptor activator of nuclear factor-κB ligand, lipopolysaccharide,Dendrobium moniliforme,Portulaca oleracea,Ampelopsis sinica,Schizonepeta".The literatures addressing osteoporosis, inflammation and herbal medicine were screened, and those using lipopolysaccharide-induced mouse models were included. Finally, four eligible literatures were enrolled for review. RESULTS AND CONCLUSION:In vivo experiments,CT images and pathological sections of the cancellous bone in the mouse distal femur show that Dendrobium moniliforme,Purslane oleracea,Ampelopsis sinica,and Schizonepeta exert inhibitory effects on lipopolysaccharide-induced osteoporosis.In vitro experiments reveal that these four kinds of herbs fight against osteoporosis by inhibiting osteoclast differentiation and further reducing bone resorption.

Puerarin prevents bone loss in ovariectomized mice and inhibits osteoclast formation in vitro / 中国天然药物

The present study aimed at investigating the effects of Puerarin (PR), a major isoflavonoid isolated from the Chinese medicinal herb Puerariae radix, on bone metabolism and the underlying mechanism of action. The in vivo assay, female mice were ovariectomized (OVX), and the OVX mice were fed with a diet containing low, middle, and high doses of PR (2, 4, and 8 mg·d(-1), respectively) or 17β-estradiol (E2, 0.03 μg·d(-1)) for 4 weeks. In OVX mice, the uterine weight declined, and intake of PR at any dose did not affect uterine weight, compared with the control. The total femoral bone mineral density (BMD) was significantly reduced by OVX, which was reversed by intake of the diet with PR at any dose, especially at the low dose. In the in vitro assay, RAW264.7 cells were used for studying the direct effect of PR on the formation of osteoclasts. PR reduced the formation of tartrate resistant acid phosphatase (TRAP)-positive multi-nucleated cells in the RAW 264.7 cells induced by receptor activator for nuclear factor-κB Ligand (RANKL). MC3T3-E1 cells were used for studying the effects of PR on the expression of osteoprotegerin (OPG) and RANKL mRNA expression in osteoblasts. The expression of OPG mRNA and RANKL mRNA was detected by RT-PCR on Days of 5, 7, 10, and 12 after PR exposure. PR time-dependently enhanced the expression of OPG mRNA and reduced the expression of RANKL mRNA in MC3T3-E1 cells. In conclusion, our results suggest that PR can effectively prevent bone loss in OVX mice without any hyperplastic effect on the uterus, and the antiosteoporosis activity of PR may be related to its effects on the formation of osteoclasts and the expression of RANKL OPG in osteoblasts.

Study on differentially expressed proteins of effect of kudiezi injection on cerebral cortexin rats with cerebral ischemic stroke and heat toxin syndrome / 中国中药杂志

This study is to investigate the modulation of Kudiezi (KDZ) injection on differential protein expression in cerebral cortex of rats with cerebral ischemic stroke and heat toxin syndrome established by intraperitoneal injection of carrageenan and middle cerebral artery occlusion (MCAO) methods. According to random number table rats were divided into three groups: drug group, model group and sham group. The tripheye tetrazolium chloride (TTC) staining and HE staining were used to observe brain tissue injury of rats. After therapeutic intervention with above drug for seventy-two hours, the level of differential protein expression was analyzed by two-dimensional gel electrophoresis (2-DE). The results show that there are differential protein expressions between cerebral ischemic stroke and heat toxin syndrome rats and sham rats. Furthermore, as a Chinese medicine injection with effect of clearing heat, resolving toxin and dredging collaterals, KDZ injection can decrease alleviate morphological changes of cerebral ischemia, regulate the levels of some differential proteins expression.

Suppressive effect in vitro of resveratrol on ADP induced human platelet aggregation and its active mechanism / 药学学报

Resveratrol (RESV) is a polyphenolic compound existed in native plants such as grape, fleeceflower root, and peanut, etc. The aim of this study was to investigate the effects in vitro of RESV on adenosine diphosphate (ADP)-induced platelet aggregation, platelet membrane-bound fibrinogen (PFig) its mechanism of action. The effects of RESV and phospholipase Cbeta inhibitor (U73122) on ADP-induced healthy human volunteers platelet aggregation, PFig, and the expression of phospho-phospholipase Cbeta3 (P-PLCbeta3) and total-phospholipase Cbeta3 (T-PLCbeta3) were studied with platelet aggregometer, flow cytometry and Western blotting, respectively. Compared with control group, RESV at 25, 50 and 100 micromol x L(-1) inhibited ADP-induced platelet aggregation and PFig in a dose dependent manner, and RESV at 25 micromol x L(-1) obviously reduced expression of P-PLCbeta3 and ratio of P-PLCbeta3 to T-PLCbeta3 in platelet of healthy human volunteers. Furthermore, RESV and U73122 had additive effect in inhibiting platelet aggregation and PFig. All these suggested that RESV inhibited platelet aggregation and PFig induced by ADP partly through decreasing the activity of PLCbeta of platelets, and that RESV had definite effect of antiplatelet and might be developed as a novel antithrombotic agent.

Cloning, expression and sequence analysis and tissue distribution of angiotensin-converting enzyme 2 (ACE2) gene in adult mice / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>o clone angiotensin-converting enzyme 2(ACE2) gene, to analyze its amino acids and nucleotides sequence and to investigate tissue distribution of ACE2 in adult mice.</p><p><b>METHODS</b>The full-length ACE2 encoding sequence was amplified from the RNA of mice kidney tissue by RT-PCR technique, cloned into plasmid pGEM-T easy, then subcloned into plasmid pcDNA3.1+. After identification of DNA sequence, the recombinant plasmid pmACE2 was transfected into Cos7 cells with lipofectin reagent. The transient expression of ACE2 molecule was detected by SDS-PAGE. Sequence analysis was conducted with CLUSTALX program. Tissue distribution of ACE2 in mice was detected by RT-PCR.</p><p><b>RESULTS</b>A fragment about 2.6 kb was amplified and the recombinant plasmid pmACE2 was confirmed by two-enzyme digesting and DNA sequencing. The cloned DNA sequence was consistent with that previously reported, except for 3 variations: A701G, T1102C and T1330C. SDS-PAGE proved that expression of a soluble, truncated products form of ACE2 was a glycoprotein of approximately 80 kD in Cos7 cells. The predicted mice ACE2 sequence contained an N-terminal signal sequence (amino acid residues 1-18), a single HHEMGHIQ zinc-binding domain (amino acid residues 373-380) and C-terminal membrane anchor (amino acid residues 738-765). Mice ACE2 showed 84 % identity with that of human, and 90 % identity with that of rat. Expression of ACE2 was the greatest in lungs, hearts and kidneys, and moderate levels were also detected in testes and livers.</p><p><b>CONCLUSION</b>Mice ACE2 gene has been cloned and successfully expressed in vitro. The tissue-specific expression of ACE2 in different species is not identical.</p>