Department of Physiology, Faculty of Science, Prince of Songkla University, Hat-Yai, Songkhla 90112, Thailand.
J Ethnopharmacol. 2013 Aug 26;149(1):123-32. doi: 10.1016/j.jep.2013.06.010. Epub 2013 Jun 15.
Tinospora crispa has been used in folkloric medicine for the control of blood pressure. We previously found that an extract of Tinospora crispa and its constituents effect the heart rate and blood pressure in anesthetized rats.
The aim was to investigate the effects and mechanisms of the Tinospora crispa extract and bioactive components on the rat isolated left atria.
Air-dried stems of Tinospora crispa were extracted with water, followed by partitioning with chloroform, ethyl acetate, and finally by n-butanol. The n-butanol soluble material was concentrated and dried under reduced pressure and lyophilized to obtain a crude powder (Tinospora crispa extract). The active components of Tinospora crispa extract were separated by column chromatography and preparative HPLC. The effects and mechanisms of the n-butanol extract and the bioactive purified components (adenine, uridine, adenosine, salsolinol, tyramine, higenamine, syringin, (-)-litcubinine, borapetoside A, borapetoside B, borapetoside D and borapetoside E) were studied in isolated left atria from normal and reserpinized rats.
Tinospora crispa extract caused an increase in the force of contraction of the electrical field stimulated left atrium. This effect was inhibited by propranolol, atenolol, ICI-118,551, phentolamine and atropine. The positive inotropic effect on the reserpenized isolated left atrium of the Tinospora crispa extract was significantly inhibited by propranolol, atenolol and ICI-118,551. Phentolamine, on the other hand, caused potentiation and the effect was inhibited when propranolol was also added. Higenamine caused an increase in the force of contraction of the electrical field stimulated left atrium and this effect was significantly inhibited by ICI-118,551 and atenolol but not by phentolamine. Reserpine did not significantly shift the concentration-response curve (C-R curve) of the inotropic effect of the higenamine. ICI-118,551 and atenolol caused a parallel shift of the C-R curve to the right of about 8 and 33 fold, respectively. At low concentrations salsolinol caused a slight increase in the force of contraction of the left atrium, but at higher concentrations a decrease was observed. The negative inotropic effect of salsolinol was significantly inhibited by propranolol and atropine. In the reserpinized isolated left atrium, the negative inotropic effect of salsolinol was potentiated and again this effect was significantly inhibited by propranolol and atropine. Tyramine caused a positive inotropic effect, and this effect was inhibited by propranolol or by pretreatment of the rat with reserpine. Adenosine caused a negative inotropic effect, while uridine caused a slight positive inotropic effect on the left atrium. This effect was significantly inhibited by DPCPX.
Crude extract of Tinospora crispa exert a positive inotropic effect on the electrical field stimulated isolated left atria that results from the concerted action of 5 bioactive compounds: higenamine, salsolinol, tyramine, adenosine and uridine. Higenamine, salsolinol (at low concentration) and tyramine acted via the adrenergic receptors to increase the force of the atrial contraction, whereas a high concentration of salsolinol acted indirectly by stimulating the release of acetylcholine. Adenosine and uridine acted via the purinergic pathways to cause negative inotropic effects on the isolated left atria.
三叶鬼针草在民间医学中被用于控制血压。我们之前发现三叶鬼针草的提取物及其成分会影响麻醉大鼠的心率和血压。
目的是研究三叶鬼针草提取物及其生物活性成分对大鼠离体左心房的作用和机制。
将三叶鬼针草的干燥茎用水提取，然后用氯仿、乙酸乙酯和正丁醇进行分配。正丁醇可溶部分浓缩并在减压下干燥，然后冻干得到粗粉（三叶鬼针草提取物）。三叶鬼针草提取物的活性成分通过柱色谱和制备 HPLC 进行分离。研究了正丁醇提取物和生物活性纯化成分（腺嘌呤、尿嘧啶、腺苷、藜芦醇、酪胺、海罂粟碱、丁香苷、（-）紫堇灵、博来霉素 A、博来霉素 B、博来霉素 D 和博来霉素 E）对正常和利血平化大鼠离体左心房的作用和机制。
三叶鬼针草提取物可引起电场刺激左心房收缩力增加。这种作用被心得安、阿替洛尔、ICI-118,551、酚妥拉明和阿托品抑制。三叶鬼针草提取物对利血平化离体左心房的正性变力作用明显被心得安、阿替洛尔和 ICI-118,551 抑制。另一方面，酚妥拉明引起增强作用，当加入心得安时，该作用被抑制。海罂粟碱可引起电场刺激左心房收缩力增加，该作用被 ICI-118,551 和阿替洛尔显著抑制，但不受酚妥拉明抑制。利血平对海罂粟碱变力作用的浓度-反应曲线（C-R 曲线）没有显著影响。ICI-118,551 和阿替洛尔使 C-R 曲线分别向右平行移动约 8 倍和 33 倍。在低浓度下，藜芦醇引起左心房收缩力略有增加，但在较高浓度下观察到收缩力下降。藜芦醇的负性变力作用被心得安和阿托品显著抑制。在利血平化离体左心房中，藜芦醇的负性变力作用被增强，再次被心得安和阿托品显著抑制。酪胺引起正性变力作用，该作用被心得安或利血平预处理的大鼠所抑制。腺苷引起负性变力作用，而尿嘧啶对左心房产生轻微的正性变力作用，该作用被 DPCPX 显著抑制。
三叶鬼针草粗提取物对电场刺激的离体左心房产生正性变力作用，这是由 5 种生物活性化合物的协同作用所致：海罂粟碱、藜芦醇、酪胺、腺苷和尿嘧啶。海罂粟碱、藜芦醇（低浓度）和酪胺通过肾上腺素能受体作用增加心房收缩力，而高浓度的藜芦醇通过刺激乙酰胆碱释放间接作用。腺苷和尿嘧啶通过嘌呤能途径对离体左心房产生负性变力作用。
