| AIMS.To explore the kinetogenic effects of serum of Semen Arecae,Dandelion,Semen Raphani and Atractylodes macrocephala on the colonic smooth muscular strip and smooth muscular cell of the rats.METHODS:1.Groups:Fifty-two Wistar rats(230±20) g were randomly divided into thirteen groups,containing 10%concentration groups(Arecae,Dandelion,Semen Raphani and Atractylodes macrocephala),50%concentration groups(Arecae,Dandelion,Semen Raphani and Atractylodes macrocephala),100%concentration groups(Arecae,Dandelion,Semen Raphani and Atractylodes macrocephala) and saline group.2.Preparation of pharmacologic serum:four rats of every group was infused by 1ml/100g two times a day for three days.With intraperitoneal anesthesia,blood was drawn from inferior vena cava after the last intragastric administration.Then pharmacologic serum was prepared.Serum of every group was about 7ml,stored at -20℃,used within 1 month。3.Observation index:contraction frequency and amplitude of colonic smooth muscular strip,notable contraction of colonic smooth muscle cell.4.Experimental methods:(1)The transverse and longitudinal smooth muscle strips were made into 7mm×3mm,placed in 5ml Kerbs liquid ventilated mixed gas(95%O2 and 5%CO2).50μl serum and positive control(1×103mol/L ACh) was added each time, contraction effect was separately observed.The result was expressed with contraction frequency and amplitude.SMUP-PC-based physiological signal processing system in real-time recorded.(2)Smooth muscle cell was isolated from the muscle layers of Wistar rat colon,referred to improved Bitar 'method.The contractile response of colonic smooth muscle cell to each serum and positive control(1×103mol/L ACh) was separately observed. The contractility was presented by the decrease of the cell length between the drug group and the control.Trypan blue staining was used to check the cell activity,the living cells was required above 90%.Results:1.vs.control serum:Serum containing 10%and 50%Semen Arecae could mostly enhance contraction frequency on the longitudinal smooth muscle strips(P＜0.05).Serum containing Dandelion of 10%concentration could enhance contraction amplitude of the transverse smooth muscle strips(P＜0.05).Serum containing Semen Raphani of 100% concentration could enhance contraction frequency of the longitudinal smooth muscle strips (P＜0.05),various concentration serum could enhance contraction amplitude on the longitudinal and transverse smooth muscle strips(P＜0.05).Serum containing Atractylodes macrocephala of 50%and 100%concentration could enhance contraction amplitude on the longitudinal smooth muscle strips(P＜0.05),various concentration serum could promote contraction frequency and amplitude on traverse smooth muscle strips(P＜0.05).2.The average length of detached cell was(69.8±7.2)μm,the average width was (8.0±3.4)μm.The average length of cell after adding 1×10-3 mol/L ACh was(50.4±4.9)μm,the average width was(9.0±4.7)μm,the length was significantly reduced.Serum of each Chinese materia medica could make dose dependent contraction at different concentration(P＜0.05).Serum containing Semen Arecae of 100%concentration had the strongest contractile effect(P＜0.05),the contraction percentage was(18±2)%.Serum containing Dandelion of 100%concentration had the strongest contractile effect(P＜0.05), the contraction percentage was(24±6%).Serum containing Semen Raphani of 100% concentration had the strongest contractile effect(P＜0.05),the contraction percentage was (24±7)%.Serum containing Atractylodes macrocephala of 100%concentration had the strongest contractile effect(P＜0.05),the contraction percentage was(23±6)%.1×10-3 mol/L ACh made notable contraction,the contraction percentage was(28±5)%.The strongest effect of each serum had no significant difference. Conclusions:1.Serum containing Semen Arecae,Herba Taraxaci,Semen Raphani and Atractylodes macrocephala can make notable kinetogenic effect on colonic smooth muscle strips in rats. The average contraction frequency and amplitude of colon longitudinal muscle strips were excited strongest by serum of 50%Semen Arecae and 50%Atractylodes macrocephala.The average contraction frequency and contraction amplitude of colon transverse muscle strips were excited strongest by serum of 10%concentration Atractylodes macrocephala.2.Serum containing Semen Arecae,Dandelion,Semen Raphani and Atractylodes macrocephala can make notable contraction on colonic smooth muscle cells in rats. |
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