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Diuretic Activity of Compatible Triterpene Components of Alismatis Rhizoma

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Diuretic Activity of Compatible Triterpene Components of Alismatis Rhizoma molecules Article Diuretic Activity of Compatible Triterpene Components of Alismatis rhizoma Xue Zhang 1,2,†, Xiao-Yan Li 1,†, Na Lin 1, Wan-Li Zhao 1, Xiao-Qiang Huang 1, Ying Chen 1, Ming-Qing Huang 1,2, Wen Xu 1,2,* and Shui-Sheng Wu 1,2,* 1 College of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China; [email protected] (X.Z.); [email protected] (X.-Y.L.); [email protected] (N.L.); [email protected] (W.-L.Z.); [email protected] (X.-Q.H.); [email protected] (Y.C.); [email protected] (M.-Q.H.) 2 Centre of Biomedical Research & Development, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China * Correspondence: [email protected] (W.X.); [email protected] (S.-S.W.); Tel.: +86-0591-2286-1622 (S.-S.W.) † These authors contributed equally to this work. Received: 15 August 2017; Accepted: 31 August 2017; Published: 6 September 2017 Abstract: Alismatis rhizoma (AR), the dried rhizoma of Alisma orientale Juzepzuk (Alismataceae), is a traditional Chinese medicine. AR is an important part of many prescriptions and is commonly used as a diuretic agent in Asia. This study aimed to evaluate the diuretic effects of total triterpene extract (TTE) and triterpene component compatibility (TCC, the mixture of alisol B 23-acetate, alisol B, alisol A 24-acetate, alisol A, and alisol C 23-acetate) of AR in saline-loaded rats. The optimal diuretic TCC of AR was optimized using a uniform design. Different doses (5, 20, and 40 mg/kg) of TTE and TCC groups (N1–N8) were orally administered to rats. Urinary excretion rate, pH, and electrolyte excretion were measured in the urine of saline-loaded rats. Results showed that TTE doses increased urine volume and electrolyte excretion compared with the control group. All uniformly designed groups of TCC also increased urine excretion. In addition, optimal diuretic TCC was calculated (alisol B 23-acetate: alisol B: alisol A 24-acetate: alisol A: alisol C 23-acetate 7.2:0.6:2.8:3.0:6.4) and further validated by saline-loaded rats. This study demonstrated that TTE presented a notable diuretic effect by increasing Na+,K+, and Cl− displacements. The most suitable TTC compatible proportion of alisol B 23-acetate: alisol B: alisol A 24-acetate: alisol A: alisol C 23-acetate for diuretic activity was validated, and triterpenes were the material basis for the diuretic activity of AR. Keywords: total triterpenes; Alismatis rhizoma; diuretic activity; component compatibility 1. Introduction In recent years, an increase in the attempts to find natural sources of molecules with biological potential has been noticed. Alismatis rhizome (AR), the dried rhizome of Alisma orientale (Sam.) Juzepzuk (Alismataceae), is a natural medicine widely cultivated in China, Japan, Korea, India, and Europe. Traditionally, AR is known to exhibit diuretic, hyperlipidemic, inflammatory, antitumoral, and damp-heat clearing actions [1–7] and has been used for treating dysuria, edema, urinary tract infection, retention of fluid and phlegm, and vertigo for more than 1000 years [8]. As a folk medicine that has long been used to promote health and longevity, AR is now included in the Pharmacopoeia of China as a representative traditional Chinese herbal diuretic. AR is an important component of many famous Chinese formulae for diuretics in the Treatise on Febrile and Miscellaneous Disease (Shang Han Lun in Chinese) including Wu Ling San, Zhu Ling Tang, and Mu Li Ze Xie San [4], for usage as diuretics due to their low toxicity, high effectiveness, and minimal side effects. Natural Molecules 2017, 22, 1459; doi:10.3390/molecules22091459 www.mdpi.com/journal/molecules Molecules 2017, 22, 1459 2 of 13 medicineMolecules is 2017 a precious, 22, 1459 resource of therapeutically active compounds including terpenoids.2 of Previous 13 studies have reported the diuretic activity of different fractions (including petroleum ether, ethyl acetate,usage n-butanol, as diuretics and due remaining to their low fractions) toxicity, ofhigh the effectiveness, ethanol extract and ofminima AR. Thel side results effects. indicated Natural that the ethylmedicine acetate is a precious (100 and resource 400 mg/kg) of therapeuticall fractionsy significantly active compounds increased including urine terpenoids. output and Previous electrolyte excretion.studies Furthermore, have reported ourthe previousdiuretic activity study of found different fourteen fractions terpenoids (including were petroleum isolated ether, from ethyl the ethyl acetateacetate, fraction n-butanol, of AR, and which remaining indicated fractions) that terpenoids of the ethanol were extract the main of AR. ingredients The results of indicated AR [9–11 that]. These the ethyl acetate (100 and 400 mg/kg) fractions significantly increased urine output and electrolyte triterpenoids (protostane-type) have attracted the most attention among the various constituents of AR, excretion. Furthermore, our previous study found fourteen terpenoids were isolated from the ethyl owingacetate to their fraction high of content AR, which and variousindicated biological that terpenoids activities were [4 ].the The main ethanol ingredients extract of ofAR AR [9–11]. and alisol A 24-acetateThese triterpenoids exerts diuretic (protostane-type) effects on animals have [attracted11], this ethanolthe most extract attention contains among mainly the various terpenoids, includingconstituents alisol Bof 23-acetate, AR, owing alisol to their B, alisolhigh Acontent 24-acetate, and various alisol A,biological and alisol activities C 23-acetate [4]. The [12 ethanol–14]. Alisol A, alisolextract B, alisolof AR A and 24-acetate, alisol A and24-acetate alisol Bexerts 23-acetate diuretic were effects the on first animals triterpenoids [11], this isolated ethanol from extract AR [15], beforecontains alisol mainly C 23-acetate terpenoids, was including also isolated alisol from B 23-acetate, AR [16 ,alisol17]. TheB, alisol most A beneficial24-acetate, alisol effects A, of and AR are attributedalisol C to 23-acetate the presence [12–14]. of terpenoidsAlisol A, alisol [12 ,B,14 alisol,18]. Therefore,A 24-acetate, we and speculated alisol B 23-acetate that these were terpenoids the first may be thetriterpenoids active compounds isolated from responsible AR [15], before for the alisol observed C 23-acetat diuretice was also effect. isolated from AR [16,17]. The most beneficial effects of AR are attributed to the presence of terpenoids [12,14,18]. Therefore, we speculated The curative effects of traditional Chinese medicine are not caused by single chemical entities, but that these terpenoids may be the active compounds responsible for the observed diuretic effect. result from their multi-ingredient prescription. The extent of compatibility depends on the integration The curative effects of traditional Chinese medicine are not caused by single chemical entities, of multitargetsbut result from and their multipathways, multi-ingredient which prescription. are mediated The extent by active of compatibility component depends compatibility on the and exertintegration corresponding of multitargets terminal effects. and multipathway Traditional Chineses, which medicine are mediated integration by active compatibility component mainly manifestscompatibility in component and exert compatibility corresponding and terminal specifically effects. relates Traditional to multicomponent Chinese medicine addition, integration competition and cooperation,compatibility andmainly antagonistic manifests action. in component Such treatment compatibility affects the and performance specifically of multicomponentrelates to multitargetsmulticomponent and shows addition, multicomponent competition and sequence cooperat amplificationion, and antagonistic and multicomponent action. Such treatment selection of a dominantaffects role.the performance of multicomponent multitargets and shows multicomponent sequence amplificationThus, the present and multicomponent study primarily selection aimed of a todominant evaluate role. the total triterpene extract (TTE) and Thus, the present study primarily aimed to evaluate the total triterpene extract (TTE) and triterpene component compatibility (TCC, including alisol B 23-acetate, alisol B, alisol A 24-acetate, triterpene component compatibility (TCC, including alisol B 23-acetate, alisol B, alisol A 24-acetate, alisol A, and alisol C 23-acetate, Figure1) of the diuretic effect of AR in saline-loaded rats and to alisol A, and alisol C 23-acetate, Figure 1) of the diuretic effect of AR in saline-loaded rats and to optimizeoptimize the the optimal optimal diuretic diuretic TCC TCC from from AR. AR. O OH OH HO OAc OH OH O HO OH HO OAc H H H O O O 1 alisol C 23-acetate 2 alisol A 3 alisol A 24-acetate O O HO OH HO OAc H H O O 4 alisol B 5 alisol B 23-acetate FigureFigure 1. Chemical 1. Chemical structures structures of of fivefive diuretic compounds compounds used used for for compatibility compatibility studies. studies. 2. Results 2. Results 2.1. Determination of Triterpenes in Total Triterpene Extract (TTE) 2.1. Determination of Triterpenes in Total Triterpene Extract (TTE) High-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS)High-performance analysis liquid showed chromatography-quadrupole that alisol A, alisol A 24-acetate, time-of-flight alisol B, alisol B mass 23-acetate, spectrometry and (HPLC-Q-TOF-MS)alisol C 23-acetate analysis were the showed major triterpenes that alisol in A,the alisol TTE (Figure A 24-acetate, 2). Table
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