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AIM:To investigate the pharmacological effect ofTong Xie-Yao Fang(TXYF)formula and its underlying mechanisms.METHODS:A neonatal maternal separation plus restraint stress(NMS+RS)model of diarrheapredominant irritable bowel syndrome was developed by subjecting male Sprague-Dawley rats to daily maternal separation from postnatal days 2 to 21 plus restraint stress from days 50 to 59.Rats were randomly divided into two groups(NMS+RS and TXYF formula),and rats with no handling or separation were used as normal controls.Starting from postnatal day 60,rats were administered TXYF formula(9.84 g/100 g body weight)orally twice daily for 14 consecutive days,while the normal and NMS+RS groups were given distilled water.The distinctions of movement index(MI,area under the curve of contraction intensity/min,mg/min)and contraction frequency(CF,number of contractions/min,times/min)of isolated colonic longitudinal smooth muscle strips(CLSMs)in the three groups before and after treatment were observed with a Power Lab system.Different inhibitors were applied,and then 10-4mol/L acetylcholine chloride(Ach)was added to CLSMs to induce muscle contraction.RESULTS:Before treatment,the MI of CLSMs in the NMS+RS and TXYF formula groups was similar and both higher than that in the normal group(545.49±73.66 mg/min vs 245.76±34.44 mg/min and551.09±54.29 mg/min vs 245.76±34.44 mg/min,P<0.01,respectively).After treatment,the MI in the TXYF formula group was lower than that in the NMS+RS group(261.39±38.59 mg/min vs 533.9±61.63 mg/min,P<0.01).In the same way,the CF of CLSMs in the NMS+RS and TXYF formula groups was similar and both higher than that in the normal group(3.42±0.25 times/min and 3.31±0.21 vs1.1±0.17 times/min,P<0.01)before treatment.After treatment,the CF in the TXYF formula group was lower than that in the NMS+RS group(1.42±0.87 times/min vs 3.11±0.82 times/min,P<0.01)and similar to that in the normal group(1.42±0.87 times/min vs 1.09±0.13 times/min).When8-(N,N-diethylamino)octyl 3,4,5-trimethoxybenzoate hydrochloride and 4-aminopyridine were added to the bath and equilibrated for 30 min,respectively,and10-4 mol/L Ach was added to CLSMs to induce muscle contraction,MI of the CLSMs in the TXYF formula group was lower than that in the normal group(666±36.32 mg/min vs 747.77±49.47 mg/min,and 686.53±39.17 mg/min vs 750.45±29.39 mg/min;P<0.01,respectively).The MI of CLSMs in the TXYF formula group was lower than that in the normal group after treatment with nifedipine(689.48±30.84 mg/min vs741.65±32.41 mg/min;P<0.05).CONCLUSION:TXYF formula inhibits colon contraction in rats.This may be related to activation of specific potassium channels and inhibition of extracellular calcium internal flow.
AIM: To investigate the pharmacological effect of Tong Xie-Yao Fang (TXYF) formula and its underlying mechanisms. METHODS: A neonatal maternal separation and restraint stress (NMS + RS) model of diarrheapredominant irritable bowel syndrome was developed by subjecting male Sprague-Dawley rats to daily maternal separation from postnatal days 2 to 21 plus restraint stress from days 50 to 59. Rats were randomly divided into two groups (NMS + RS and TXYF formula), and rats with no handling or separation were used as normal controls. Starting from Postnatal day 60, rats were administered TXYF formula (9.84 g / 100 g body weight) orally twice daily for 14 consecutive days, while the normal and NMS + RS groups were given distilled water. The distinctions of movement index (MI, area under the curve of contraction intensity / min, mg / min) and contraction frequency (CF, number of contractions / min, times / min) of isolated colonic longitudinal smooth muscle strips (CLSMs) in the three groups before and after treatment were observed wi th a Power Lab system. Different inhibitors were applied, and then 10-4 mol / L acetylcholine chloride (Ach) was added to CLSMs to induce muscle contraction .RESULTS: Before treatment, the MI of CLSMs in the NMS + RS and TXYF formula groups was similar and both higher than that in the normal group (545.49 ± 73.66 mg / min vs 245.76 ± 34.44 mg / min and 551.09 ± 54.29 mg / min vs 245.76 ± 34.44 mg / min, P <0.01, respectively) .After treatment , the MI in the TXYF formula group was lower than that in the NMS + RS group (261.39 ± 38.59 mg / min vs 533.9 ± 61.63 mg / min, P <0.01) .In the same way, the CF of CLSMs in the NMS + RS and TXYF formula groups were similar and both higher than that in the normal group (3.42 ± 0.25 times / min and 3.31 ± 0.21 vs 1.1 ± 0.17 times / min, P <0.01) before treatment. After treatment, the CF in the TXYF formula group was lower than that in the NMS + RS group (1.42 ± 0.87 times / min vs 3.11 ± 0.82 times / min, P <0.01) and similar to that in the normal group (1.42 ± 0.87 times / min vs 1.09 ± 0.13 times / min) .When8- (N, N-diethylami no) octyl 3,4,5-trimethoxybenzoate hydrochloride and 4-aminopyridine were added to the bath and equilibrated for 30 min, respectively, and 10-4 mol / L Ach was added to CLSMs to induce muscle contraction, MI of the CLSMs in the TXYF formula group was lower than that in the normal group (666 ± 36.32 mg / min vs 747.77 ± 49.47 mg / min and 686.53 ± 39.17 mg / min vs 750.45 ± 29.39 mg / min; P <0.01, respectively). The MI of CLSMs in the TXYF formula group was lower than that in the normal group after treatment with nifedipine (689.48 ± 30.84 mg / min vs 741.65 ± 32.41 mg / min; P <0.05) .CONCLUSION: TXYF formula inhibits colon contraction in rats. This may be related to activation of specific potassium channels and inhibition of extracellular calcium internal flow.