Coptis Chinensis Franch And Radix Ophiopogonis Extract On Rats Conjunctival Goblet Cell In Vitro Proliferation Function And Effective Ingredients Compatibility Screening Of Experimental Study

Purpose:(1) Establish SD Rats Conjunctival Goblet cells (Rats Conjunctival Goblet Cells, RCj-GCs) in vitro conventional cultivating model; (2) Observation coptidis and dwarf lilyturf five extract intervene RCj-GCs 48h the cell proliferation activity; (3) Observation compatibility of effective ingredients intervene RCj-GCs 24h and 48h respectively the cell proliferation activity; (4) Observation Compatibility group intervene RCj-GCs 48h cell cycle effects; (5) Observation Compatibility group intervene RCj-GCs 48h for cellular proteins secreted effects.Methods:(1) Tissue mass primary culture cells, characterization cells by histochemistry and Immunohistochemical; (2) Will five experimental drug:Coptis polysaccharide (CP), coptidis alkaloids (CA), Dwarf lilyturf polysaccharides (DLP), Dwarf lilyturf isoflavone (DLI), Dwarf lilyturf saponins (DLS) dissolved and dilute respectively for 8 the experimental group, final concentration respectively is 10mg/ml, 5 mg/ml,1 mg/ml,0.2 mg/ml,0.04 mg/ml,0.008mg/ml,0.0016 mg/ml,0.00032 mg/ml; detecting 8 group intervention RCj-GCs 48h proliferative activity by MTT method; (3) detecting 4 compatibility groups respectively intervention RCj-GCs 24h and 48 cell proliferative activity by MTT method;(4)detecting 4 compatibility groups cell cycle by Flow cytometric analysis; (5) detecting 4 compatibility groups the cell protein secretion culture solution by Coomassie brilliant blue G-250 (CBBG).Results:(1)With the tissue mass primary culture.in 7-10 days can cultivate RCj-GCs, Identified as RCj-GCs by AB/PAS and immunohistochemistry; (2) CP1 mg/ml, DLI 1 mg/ml and DLP0.00032 mg/ml intervention RCj-GCs proliferative activity significantly higher than in control group (P＜0.05); (3) Compatibility groups of the intervention RCj-GCs 24h proliferation activity was significantly higher than negative control group and the positive control group (P<0.01); Compatibility groupsⅠ,ⅢandⅣon RCj-GCs interfere with the proliferation activity of 48h was significantly higher than the negative control group (P<0.01); Compatibility groupsⅠon RCj-GCs intervention the proliferation activity of 48h was significantly higher than the positive control group (P＜0.01); Compatibility groupⅢon RCj-GCs interfere with the proliferation activity of 48h was significantly higher than the positive control group (P＜0.05); (4) There was no significant different compatibility group on RCj-GCs promote apoptosis; compatibility groupⅠG2/M phase and compatibility groupⅢS phase cells is higher than the negative control grou (P＜0.05); compatibility GroupⅠS phase cells is higher than the negative control group (P＜0.01); compatibility groupⅠG2/M phase cells is higher than the positive control group (P＜0.05) (5) The compatibility group intervention RCj-GCs48h cell mucin secretion compared with control group no significant difference (P>0.05).Conclusion:(1) Successfully established RCj-GCs in vitro culture model of normal, and identified as RCj-GCs; (2) CP1mg/ml, DLI1mg/ml and DLP 0.00032mg/ml on RCj-GCs have a good effect on the proliferation; (3) Compatibility groupⅠandⅢpromoted the RCj-GCs proliferation obviously; (4) Compatibility groupⅠandⅢcan promote cell division and proliferation; (5) Compatibility of each group of cells no inhibition of mucin secretion.