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ObjectiveTo investigate the mechanism of ethyl acetate extract of Tibetan medicine dampness bud Gentianopsis paludosa in the prevention and treatment of recurrent ulcerative colitis （UC） in rats with dampness-heat in large intestine syndrome based on the apoptotic pathway mediated by the B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax）. MethodUsing the disease-syndrome combination method， a recurrent UC model of dampness-heat in large intestine syndrome was constructed in rats. Seventy SPF-grade male SD rats were randomly divided into control group， model group， high-， medium-， and low-dose ethyl acetate of G.paludosa groups （150， 75， 37.5 mg·kg-1）， and mesalazine group （135 mg·kg-1）. The rats were orally administered with respective drugs for 14 days. The general conditions of the rats were recorded， and colon length and mucosal damage were observed. The colon wet weight index and organ coefficients of the liver， spleen， and thymus were calculated. Enzyme-linked immunosorbent assay （ELISA） was used to measure the levels of interleukin-6 （IL-6） and interleukin-1β （IL-1β） in the serum of each group. Hematoxylin-eosin （HE） staining was performed to observe pathological changes in the colon. Terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） was used to detect apoptosis in colonic epithelial cells. Western blot was used to measure the expression levels of Bcl-2， Bax， Caspase-3， Caspase-9， Zona Occludens-1 （ZO-1）， Claudin3， and Occludin in colonic tissue. Immunohistochemistry （IHC） was used to observe the expression of Bax and Caspase-3 in colonic epithelial cells. ResultCompared with the control group， the model group showed significant increases in the disease activity index （DAI） score， colonic mucosal damage index （CMDI）， intestinal epithelial apoptosis， liver and spleen indexes， and levels of inflammatory factors IL-1β and IL-6 in the serum （P<0.01）， decreased expression of intestinal mucosal protective proteins ZO-1， Claudin3， and Occluding （P<0.01）， increased expression of pro-apoptotic proteins Bax， Caspase-3， and Caspase-9 （P<0.01）， and decreased expression of anti-apoptotic protein Bcl-2 （P<0.01）. Compared with the model group， the high-， medium-， and low-dose ethyl acetate of G.paludosa groups all significantly improved the general condition of the rats， reduced colonic lesions， decreased intestinal epithelial cell apoptosis， reduced liver and spleen indexes， upregulated the expression of ZO-1， Claudin3， Occludin， and Bcl-2 proteins， and downregulated the expression of Bax， Caspase-3， and Caspase-9 proteins， with the high- and medium-dose ethyl acetate of G.paludosa groups showing the superior effects （P<0.05， P<0.01）. ConclusionEthyl acetate of G.paludosa can alleviate colonic mucosal damage and exert a therapeutic effect on UC by regulating the Bcl-2/Bax signaling pathway.
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Objective To compare analgesic and anti-inflammatory effects of Gentianopsis Paludosa and Sophora Alopecuroide of different kinds of compatibility on the mice.Methods Mice were randomly divided into normal control group, positive medicine group, and different kinds of compatibility groups, 10 mice in each group. Mice received gavage for successive 7 days, once a day. Glacial acetic acid twist body method and hot board to pain method were used to observe and compare analgesic effects of different kinds of compatibility. Xylene to ear swelling method and agar to granulation swollen method were used to observe and compare inflammatory effects of different kinds of compatibility.Results Compared with normal group, compound Gentianopsis Paludosa had obvious analgesic and anti-inflammatory effects (P<0.01). When the ratio of Gentianopsis Paludosa and Sophora Alopecuroide was 20:0.6 and 20:0.9, the analgesic and anti-inflammatory effects were the best.Conclusion Gentianopsis Paludosa and Sophora Alopecuroide have good analgesic and anti-inflammatory effects, among which, when the ratio of Gentianopsis Paludosa and Sophora Alopecuroide is 20:0.6, the best effects show.
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Objective To establish the determination of luteolin and 1,7-dihydroxy-3,8-dimethoxyxanthone in Herbal Gentianopsis paludosa. Method The RP-HPLC with ZORBAX SB-C18 (250 mm?4.6 mm, 5 ?m) comlum was used. Mobile phase was methanol (A)-0.4% H3PO4 (B) Gradient elution was used, 0～15 min, A∶B=50∶50, 15～60 min, A∶B=55∶45. Detection wavelengh was 260 nm. Flow rate was 1.0 mL/min. Temperature of column was 30 ℃. Result The linear relationship of 1,7-dihydroxy-3,8-dimethoxyxanthone and luteolin were showed at the range of 0.084～0.84 ?g and 0.184～1.84 ?g (r=0.999 4 and r =0.999 7). The average recovery rate of 1,7-dihydroxy-3,8-dimethoxyxanthone and luteolin were 96.82% and 97.19%, RSD