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AIM:Menoease Pills(MP),a Chinese medicine-based newformula for postmenopausal women,has been shown tomodulate the endocrine and immune systems.The presentstudy investigated the effects of MP and one of its activeingredients,ligustrazine,on epithelial barrier and ion transportfunction in a human colonic cell line,T_(84).METHODS:Colonic transepithelial electrophysiologicalcharacteristics and colonic anion secretion were studied usingthe short circuit current(I_(SC))technique.RT-PCR was used toexamine the expression of cytoplasmic proteins associatedwith the tight junctions,ZO-1(zonula occludens-1)and ZO-2(zonula occludens-2).RESULTS:Pretreatment of T_(84) cells with MP(15 μg/mL)for72 h significantly increased basal potential difference,transepithelial resistance and basal I_(SC).RT-PCR results showedthat the expressions of ZO-1 and ZO-2 were significantlyincreased after MP treatment,consistent with improvedepithelial barrier function.Results of acute stimulationshowed that apical addition of MP produced a concentration-dependent(10-5 000 μg/mL,EC_(50)=293.9μg/mL)increasein I_(SC).MP-induced I_(SC) was inhibited by basolateral treatmentwith bumetanide (100 μmol/L),an inhibitor of the Na~+-K~+-2CI~- cotransporter,apical addition of CI-channelblockers,diphenylamine-2,2’-dicarboxylic acid(1 mmol/L)orglibenclamide(1 mmol/L),but not 4,4’-diisothiocyanostilbene-2,2’-disulfonic acid or epithelial Na+ channel blocker,amiloride.The effect of MP on ZO-1 and ZO-2was mimickedby Ligustrazine and the ligustrazine-induced I_(SC) was alsoblocked by basolateral application of bumetanide andapical addition of diphenylamine-2,2’-dicarboxylic acid orglibenclamide,and reduced by a removal of extracellular CI~-.CONCLUSION:The results of the present study suggestthat MP and Iligustrazine may improve epithelial barrierfunction and exert a stimulatory effect on colonic anionsecretion,indicating the potential use of MP and its activeingredients for improvement of GI tract host defense andalleviation of constipation often seen in the elderly.
AIM: Menoease Pills (MP), a Chinese medicine-based newformula for postmenopausal women, has been shown in tomodulate the endocrine and immune systems. The present study examines the effects of MP and one of its active ingredients, ligustrazine, on epithelial barrier and ion transport function in a human colonic cell line, T 84 (84). METHODS: Colonic transepithelial electrophysiological characteristics and colonic anion secretion were studied using the short circuit current (I SC) technique. RT-PCR was used toexamine the expression of cytoplasmic proteins associated with the tight junctions, Prevalence of T_ (84) cells with MP (15 μg / mL) for 72 h of significantly increased basal potential difference, transepithelial resistance and basal I_ (Zonula occludens-1) and ZO- (SC). RT-PCR results showed that the expressions of ZO-1 and ZO-2 were significantly increased after MP treatment, consistent with improvedepithelial barrier function. Results of acute stimulation showed that apical additi (SC) was inhibited by basolateral treatment with bumetanide (100 μmol / L) on of MP produced a concentration-dependent (10-5 000 μg / mL, EC 50 = 293.9 μg / L, an inhibitor of the Na ~ + -K ~ + -2CI ~ - cotransporter, apical addition of CI-channel blockers, diphenylamine-2,2’- dicarboxylic acid (1 mmol / L) orglibenclamide but not 4,4’-diisothiocyanostilbene-2,2’-disulfonic acid or epithelial Na + channel blocker, amiloride. The effect of MP on ZO-1 and ZO-2 was mimicked by Ligustrazine and the ligustrazine-induced I SC was also blocked by basolateral application of bumetanide andapical addition of diphenylamine-2,2’-dicarboxylic acid orglibenclamide, and reduced by a removal of extracellular CI ~ -CONCLUSION: The results of the present study suggest that MP and Iligustrazine may improve epithelial barrier function and exert a stimulatory effect on colonic anionsecretion, indicating the potential use of MP and its activeingredients for improvement of GI tract host defense andal leviation of constipation often seen in the elderly.