| Objective:To observe the influence of serum containing Zhi Gancao Decoction on the electrophysiologyical effect in isolated rabbit's ventricular myocytes.Methods: (1) First make the blood serum containing Zhi Gancao Decoction. (2) The glycyrrhetinic acid concentration was determined by RP-HPLC with methanol-water-acetic acid (85 : 14 : 1) as mobile phase,while Hypersil BDS C18 column(4.6mm×200mm, 5um)as stationary phase, and detected at UV(λ=250nm). (3) The influence of normal serum on the APD in isolated rabbit's ventricular myocytes : Single rabbit's ventricular myocytes were isolated with enzyme. The whole-cell patch clamp recording technique was used to observe the changes of APD. The study was divided into five consist of groups : normal group, 5%,10%, 20%, 40% serum group. (4) The influence of serum containing Zhi Gancao Decoction on the APD in isolated rabbit's ventricular myocytes : Single rabbit's ventricular myocytes were isolated with enzyme. The whole-cell patch clamp recording technique was used to observe the changes of APD. The study was divided into six consist of groups : normal group , normal serum group, 5%, 10%, 20%, 40% serum containing Zhi Gancao Decoction group. (5) The influence of serum containing Zhi Gancao Decoction on the ICa-L in isolated rabbit's ventricular myocytes : Single rabbit's ventricular myocytes were isolated with enzyme. The whole-cell patch clamp recording technique was used to observe the changes of ICa-L The study was divided into eight consist of groups : normal group , normal serum group, 5%,10%, 20%, 40% serum containing Zhi Gancao Decoction group,verapamil group, verapamil and serum group. (6) The influence of serum containing Zhi Gancao Decoction on the Ito in isolated rabbit's ventricular myocytes : Single rabbit's ventricular myocytes were isolated with enzyme. The whole-cell patch clamp recording technique was used to observe the changes of Ito. The study was divided into six consist of groups : normal group , normal serum group, 5%,10%, 20%, 40% serum containing Zhi Gancao Decoction group. (7) The influence of serum containing Zhi Gancao Decoction on the INa in isolated rabbit's ventricular myocytes : Single rabbit's ventricular myocytes were isolated with enzyme. The whole-cell patch clamp recording technique was used to observe the changes of INa. The study was divided into six consist of groups : normal group , normal serum group, 5%,10%, 20%, 40% serum containing Zhi Gancao Decoction group.Results: (1) After oral Zhi Gancao Decoction, the blood drug concentration peak time is about 1.5 hour. (2) The normal serum can't prolong the APD in isolated rabbit's ventricular myocytes. (3) The serum containing Zhi Gancao Decoction prolonged the APD in isolated rabbit's ventricular myocytes, 5%,10%,20%, 40% serum group prolonged APD50 from (198.8±35.9)ms to (352.7±39.2)ms, (430.1 ± 28.8)ms,(521.2 ± 35.3)ms,(539.3 ±26.8)ms, and prolonged APD90 from (340.1 ±32.9) ms to (447.1±26.8) ms,(526.0±39.4) ms, (595.7 ± 53.3) ms, (624.3± 54.0) ms. (4) The serum containing Zhi Gancao Decoction decreased the Ica-L density in isolated rabbit's ventricular myocytes, 5%, 10%, 20%, 40% serum group decreased the ICa-L density from (9.4 ± 1.0) PA/PF to (7.8 ±1.0) PA/PF, (6.7 ± 0.7) PA/PF, (5.6 ± 0.9) PA/PF, (5.7±1.1) PA/PF, there were no remarkable difference between 20%, 40% serum group and verapamil group, verapamil serum group. (5) The serum containing Zhi Gancao Decoction decreased the Ito density in isolated rabbit's ventricular myocytes, 5%, 10%, 20%, 40% serum group decreased the Ito density from (16.1 ± 1.4) PA/PF to (13.9±1.5) PA/PF, (11.8±1.9) PA/PF, (8.3 ± 1.5) PA/PF,( 8.2 ±1.2 ) PA/PF. (6) The serum containing Zhi Gancao Decoction can't decrease the INa density.Conclusion: (1) After oral Zhi Gancao Decoction, the blood drug concentration peak time is about 1.5 hour. (2) The serum containing Zhi Gancao Decoction prolonged the APD in isolated rabbit's ventricular myocytes,and decreased the ICa-L, Ito density.So it can act on severa... |
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