ABSTRACT
OBJECTIVE To study the improving mechanism of Buyang huanwu decoction （BYHW） on idiopathic pulmonary fibrosis （IPF） model rats. METHODS Ninety-six Wistar rats were randomly divided into blank group， model group， positive control group， and BYHW low-dose， middle-dose and high-dose groups， with 16 rats in each group. Except for the blank group， the IPF model was induced in other groups by intratracheal instillation of bleomycin （5 mg/kg）. Starting from the day of modeling， the blank group and model group were given normal saline （10 mL/kg） intragastrically， while the rats in the positive control group were given dexamethasone solution （5 mg/kg） intragastrically， BYHW low-dose， middle-dose and high-dose groups were treated with BYHW （2.5， 5， 10 g/kg， by crude drug） intragastrically， once a day， for 28 consecutive days. The body mass of rats in each group was measured on days 0， 7， 14， 21 and 28 after modeling. The number of adherent white blood cells in pulmonary veins was observed by a dynamic visualization system. The contents of TNF-α and IL-6 in serum and TNF-α， IL-6， HYP and TGF-β1 in lung tissue were detected； the protein expression of ZO-1 was also detected. The pathomorphological changes in lung tissue were observed. RESULTS Compared with the model group， the body weight of rats all increased significantly in BYHW high-dose and middle-dose groups， positive control group， while the number of adherent white blood cells in pulmonary veins was decreased significantly； the contents of TNF-α （except for serum in BYHW middle-dose group） and IL-6 in serum and lung tissue， the contents of HYP and TGF-β1 in lung tissue were decreased significantly， while the protein expression of ZO-1 in the lung tissue was increased significantly （P＜0.05 or P＜0.01）. The pathological changes of lung tissue were improved to varying degrees. CONCLUSIONS BYHW may play anti-pulmonary fibrosis role by improving leukocyte adhesion， anti-inflammatory， anti-fibrosis， and other aspects of pulmonary microcirculation.
ABSTRACT
ObjectiveTo study the mechanism of Danggui Sinitang in mitigating gouty arthritis （GA） in rats by regulating autophagy via the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway. MethodSixty male SD rats were randomly assigned into normal， model， colchicine （0.3 mg·kg-1）， and low-， medium-， and high-dose Danggui Sinitang （6.54， 13.08， and 26.16 g·kg-1） groups （n=10） and administrated with corresponding drugs by gavage. The rats in the normal group and model group were administrated with equal volume of normal saline by gavage for 7 days. One hour after administration on day 5， the GA model was established by injecting sodium urate suspension （50 g·L-1） into the right ankle joint of rats in other groups except the normal group， and the rats in the normal group were injected with sterile normal saline of the same volume. The swelling and pathological changes of the ankle joint were observed. The serum levels of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and IL-1β were determined. Western blot was employed to determine the protein levels of PI3K， phosphorylated PI3K （p-PI3K）， protein kinase B （Akt）， phosphorylated Akt （p-Akt）， mTOR， phosphorylated mTOR （p-mTOR）， microtubule-associated protein 1 light chain 3 Ⅱ/Ⅰ （LC3Ⅱ/Ⅰ）， autophagy effector Beclin-1， and ubiquitin-binding protein p62 in the synovial tissue. Real-time fluorescent quantitative PCR （Real-time PCR） was employed to determine the mRNA levels of PI3K， Akt， mTOR， LC3， Beclin-1 and p62. ResultCompared with the normal control， the model group showed increased joint swelling index （P<0.01）， elevated serum levels of TNF-α， IL-6， and IL-1β， inflammatory cell infiltration， and fibrous tissue hyperplasia. In addition， the model group showed up-regulated protein levels of PI3K， p-PI3K， Akt， p-Akt， mTOR， p-mTOR， and p62 and mRNA levels of PI3K， Akt， mTOR， and p62 in the synovial tissue， while it showed down-regulated protein levels of LC3Ⅱ/Ⅰ and Beclin-1 and mRNA levels of LC3 and Beclin-1 （P<0.01）. Compared with the model group， medium- and high-dose Danggui Sinitang alleviated the joint swelling （P<0.01）， lowered the serum levels of TNF-α， IL-6， and IL-1β （P<0.05）， and relieved the inflammatory cell infiltration in the synovial tissue of the ankle joint and the fibrous tissue hyperplasia. Moreover， they down-regulated the protein levels of PI3K， p-PI3K， Akt， p-Akt， mTOR， p-mTOR， and p62 and the mRNA levels of PI3K， Akt， mTOR， and p62 in the synovial tissue （P<0.05）， while they up-regulated the protein levels of LC3Ⅱ/Ⅰ and Beclin-1 and the mRNA levels of LC3 and Beclin-1 （P<0.05）. ConclusionDanggui Sinitang， especially at a high dose， can inhibit PI3K/Akt/mTOR signaling pathway to improve autophagy in the synovial tissue， thereby mitigating GA.
ABSTRACT
The HPLC-ELSD method was used in the content determination of astragaloside, astragalosideⅠ, astragalosideⅡand astragalosideⅢ in Mongolia Radix Astragali (Astragalus membranaceus(Fisch.) Bge. var. mongholicus(Bge.) Hsiao) among 16 batches from various habitats. The DIKMA Diamonsil C18 (150 mm× 4.6 mm, 5μm) was adopted with acetonitrile and water as the mobile phase at a gradient mode program. The flow rate was 1.0 mL·min-1. And the column temperature was 30℃. The ELSD detector parameters were the drift tube temperature at 90℃, and the air flow rate of 2.8 L·min-1. The SPSS 16.0 software was used in the cluster analysis of content determination. The results showed that when the injection volume was within the range of 0.093 2-1.02μg (r = 0.999 5), 0.789-8.78μg (r = 0.999 7), 0.506-3.13μg (r = 0.999 6), and 0.016 1-1.38μg (r = 0.999 2) for astragaloside, astragalosideⅠ, astragalosideⅡ and astragalosideⅢ, respectively, the average recoveries were 97.55%, 98.61%, 99.68%, 98.58%with RSD of 1.2%, 1.3%, 1.3%, 1.2%, respectively. The results of cluster analysis showed that the single using of astragaloside as index was unable to differentiate Mongolia Radix Astragali from various habitats. However, the simultaneous determination of 4 types of astragalosides as indexes can differentiate Mongolia Radix Astragali from various habitats. It was concluded that the method was simple, quick and accurate, which can directly reflect the quality status of Mongolia Radix Astragali from different origins. It also provided new ideas for the quality control of Mongolia Radix Astragali.