罗汉果细胞悬浮培养体系的建立与优化

目的:研究6-BA,萘乙酸(NAA),碳源,pH对罗汉果悬浮培养细胞生长的影响,探讨罗汉果细胞悬浮培养的最适条件.方法:考察1个培养周期内罗汉果悬浮细胞的鲜重和干重,绘制生长曲线和生长速率曲线;采用正交试验设计对6-BA,NAA,碳源,pH 4因素3水平的组合进行优化.结果:罗汉果悬浮细胞培养的生长周期约为21 d,0～6 d为细胞的延滞期,7～21 d为对数期,稳定期较短,第22天进入到衰亡期,在第21天时细胞鲜、干质量达到最大,分别为693.0,416.0 g、L-1,最大生长速率为0.040 g·L-1·d-1.NAA是影响罗汉果悬浮细胞生长的关键因素,其影响效应依次为NAA＞6-BA＞pH＞碳源.悬浮细胞生长的适宜培养基为MS+1.0 mg·L-1 6-BA +0.02 mg·L-1 NAA,4.0％蔗糖为碳源,初始培养基pH5.5～6.0.结论:通过组织培养方式可以成功建立罗汉果细胞悬浮培养体系.     

罗汉果甜苷对PC12细胞氧化应激的保护作用研究

目的 考察罗汉果甜苷在PC12细胞中对外源性氧化损伤的保护作用.方法 采用MTT法考察0.01 ～ 100μg/ml罗汉果甜苷对正常PC12细胞活力的影响,并在H2O2刺激的PC12细胞中考察0.01 ～ 10 μg/ml的罗汉果甜苷对细胞活力的影响,通过对细胞活力的考察选择罗汉果甜苷合适的实验剂量,测定罗汉果甜苷0.1及1μg/ml对H2O2刺激的PC12细胞中过氧化氢酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活力的影响以及对丙二醛(MDA)含量的影响.结果 罗汉果甜苷在1 ～ 10 μg/ml浓度能提高正常PC12细胞活力,在0.1～ 10 μg/ml浓度下能显著提高H2O2刺激的PC12细胞活力(P＜0.01).0.1及1μg/ml两个剂量的罗汉果甜苷均能显著提高H2O2刺激的PC12细胞中SOD活力(P＜0.001)和GSH-Px活力(P ＜0.001),并减少MDA生成(P＜0.05).结论 罗汉果甜苷能缓解氧化应激导致的PC12细胞活力下降,并能增强PC12细胞的抗氧化能力.     

罗汉果甜苷对体外诱导肝细胞脂肪变性的影响

目的 探讨罗汉果甜苷对体外诱导肝细胞脂肪变性的影响.方法 以人肝L-02细胞作为研究对象,用四环素建立肝细胞脂肪变性模型.将实验分为诱导组、继续诱导组、脱离诱导组,用油红O染色观察细胞形态和细胞内脂滴的形成情况,并测定细胞内甘油三酯(TG)含量的变化.结果 罗汉果甜苷(20～80μg·ml-1)可明显降低细胞内TG值,并呈剂量依赖性.结论 罗汉果甜苷通过减少细胞内TG的生成从而阻止肝细胞脂肪变性.     

罗汉果甜苷对体外人肝星状细胞活化和凋亡的影响

目的研究罗汉果甜苷对体外培养人肝星状细胞LX-2活化及凋亡的影响。方法体外培养LX-2,将其分为空白对照组、不同质量浓度的罗汉果甜苷干预组(80、160、240、320μg/mL),每组8瓶细胞,药物与细胞共同孵育24 h后,采用CCK-8试剂检测细胞增殖状态,用酶联免疫吸附法测定转化生长因子-β1(TGF-β1)和Ⅰ型胶原的蛋白分泌,用RT-PCR法检测TGF-β1和α-平滑肌肌动蛋白(α-SMA)的mRNA的表达。用流式细胞仪双染法检测细胞凋亡。结果 CCK8结果显示罗汉果甜苷各剂量组作用LX-2细胞24 h后,细胞增殖均明显降低(P<0.05);ELISA和RT-PCR结果表明,与空白对照组比较,罗汉果甜苷各剂量组可抑制LX-2中TGF-β1和Ⅰ型胶原的蛋白分泌;并能抑制TGF-β1和α-SMA的mRNA表达(P<0.05)。流式细胞检测结果显示罗汉果甜苷各剂量组亦可剂量依赖性显著增加LX-2凋亡率。结论罗汉果甜苷可通过抑制TGF-β1和I型胶原的蛋白及TGF-β1和α-SMA的mRNA的表达来抑制肝星状细胞活化,还能促进肝星状细胞凋亡,从而发挥抗肝纤维化的作用。     

罗汉果甜苷对肝星状细胞HSC-T6增殖及肝纤维化相关基因的影响

目的 研究罗汉果甜苷含药血清对大鼠肝星状细胞HSC-T6的毒性以及对细胞增殖与相关基因表达的影响,初步探讨其抗肝纤维化作用机制.方法 以罗汉果甜苷含药血清干预HSC-T6,乳酸脱氢酶(LDH)法检测罗汉果甜苷含药血清对HSC-T6的细胞毒性;MTT法检测其对HSC-T6生长的抑制作用;RT-PCR法检测HSC-T6中Ⅰ型胶原(Col-Ⅰ)、转化生长因子-β1 (TGF-β1)及基质金属蛋白酶抑制因子-1(TIMP-1)mRNA表达.结果 罗汉果甜苷含药血清对HSC-T6无特异毒性,罗汉果甜苷含药血清体积分数为20％、10％时可显著抑制HSC-T6的增殖(P＜0.05、0.01),并对Col-Ⅰ、TGF-β1及TIMP-1mRNA表达均表现显著抑制作用(P＜0.05、0.01).结论 罗汉果甜苷含药血清对HSC-T6细胞无特异毒性,对HSC-T6细胞增殖、Col-Ⅰ有抑制作用,促进细胞外基质(ECM)降解,这可能是其抗肝纤维化的部分作用机制.     

不同罗汉果品种生育后期净光合速率变化的比较研究

目的:分析净光合速率(P_n)与罗汉果品质的关系.方法:以永青一号和野红一号2个生育后期的罗汉果品种为实验材料,采用CI-310便携式光合作用测定系统测生育后期的净光合速率;用HPLC测甜苷V.结果:永青一号的光补偿点低于野红一号,光饱和点相同.在整个生育后期永青一号的Pn、干果重和甜苷V含量均高于野红一号.结论:净光合速率可作为评价罗汉果品质的重要辅助指标.     

罗汉果甜苷的提取与药理作用研究概况

罗汉果[Siraitia grosvenorii(Swingle)C.Jeffrey]为我国传统保健药材,是一种极具开发潜力的天然产物资源,罗汉果甜苷是罗汉果中主要有效成分,具有广泛的生物特性,其中罗汉果苷Ⅴ为罗汉果果实中含量和甜度均较高的成分,其含量约为1%,甜度相当于蔗糖的350倍,是主要的甜味成分[1]     

罗汉果甜苷保肝作用实验研究

目的:研究罗汉果甜苷(mogroside,Mog)的保肝作用。方法:将72只小鼠随机分成6组:正常对照组、模型组、联苯双酯(BPD)组(0.6 g.kg-1)、Mog高、中、低剂量组(1.5,0.75,0.38 g.kg-1)。除正常组和模型组外,其余小鼠每日ig给药1次,共7 d,末次给药1 h后各给药组及模型组腹腔注射(ip)0.08%CCl41次(10 mL.kg-1)。造模后12 h,采血测定血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)含量,并观察肝组织病理变化;另取72只Wistar大鼠,随机分成6组:正常对照组、模型组、联苯双酯(BPD)组(0.1 g.kg-1)、Mog高、中、低剂量组(0.8,0.4,0.2 g.kg-1)。除正常对照组外,其他大鼠按2 mL.kg-1(首次5 mL.kg-1)皮下注射(sc)25%CCl4每周2次,连续8周,造成慢性肝损伤,观察罗汉果甜苷对大鼠血清ALT,AST活性,透明质酸(HA)、Ⅲ型前胶原氨基端肽(PⅢNP)、羟脯氨酸(Hyp)含量,肝组织超氧化物歧化酶(SOD)、谷胱甘肽(GSH-Px)活性及丙二醛(MDA)含量的影响,观察罗汉果甜苷对肝组织转化生长因子β1表达的影响及对肝组织病理变化的影响。结果:罗汉果甜苷对CCl4诱导小鼠急性肝损伤,可降低血清中ALT,AST活性,明显减轻肝组织病理变化程度;对CCl4所致大鼠慢性肝损伤,罗汉果甜苷可降低血清中ALT,AST活性;降低HA,PⅢNP,HyP含量;升高肝组织SOD,GSH-Px活性;降低肝组织MDA含量;抑制肝组织转化生长因子β1(TGF-β1)的表达,并明显减轻肝组织病理变化程度。结论:罗汉果甜苷对CCl4所致小鼠急性肝损伤有保护作用;对CCl4所致大鼠慢性肝损伤有防治作用,并有一定的抗肝纤维化作用。     

喜树胚轴细胞悬浮培养的优化和喜树碱的积累

目的优化喜树胚轴细胞悬浮培养体系生产喜树碱。方法将由喜树种子下胚轴诱导的愈伤组织进行悬浮培养,研究了基本培养基、激素和培养条件对培养物生长的影响,测定了培养细胞生长和喜树碱积累的动态变化。结果悬浮培养细胞在M S培养基中细胞生长良好,细胞分散性和喜树碱积累优于在其他培养基中。最佳激素组合为2,4-D 0.2 m g/L NAA 0.5 m g/L 6-BA 0.5 m g/L,细胞继代周期以12 d左右为宜,悬浮细胞适合在22~26℃,120 r/m in下生长。悬浮培养细胞在第4~20天为对数生长期,细胞倍增时间为t(d)=3.92-d 1,细胞干重至第20天达到最大,为27.44 g/L。在细胞对数生长期内,喜树碱积累与细胞生长呈线形正相关性,第20天喜树碱质量分数达到最大值9.319×10-5,喜树碱产量为2.56 m g/L。结论M S基本培养基适合喜树胚轴细胞悬浮培养,细胞生长前20天内,培养体系中喜树碱积累与细胞生长呈正线形相关性。     

HPLC法测定罗汉果中罗汉果苷V和11-氧化罗汉果苷V

目的建立HPLC法测定不同产地、不同栽培品系的罗汉果药材中罗汉果苷V和11-氧化罗汉果苷V。方法采用ODS(250mm×4.6mm,5μm)色谱柱,以乙腈-水为流动相进行梯度洗脱,体积流量0.75mL/min,检测波长210nm,柱温40℃。结果罗汉果苷V在0.8046~20.1150μg线性关系良好(r=0.9998),平均回收率为104.6%,RSD为3.28%(n=6)。11-氧化罗汉果苷V在0.5985~14.9625μg线性关系良好(r=0.9984)。结论本方法简便、快速、准确,可作为罗汉果药材的质量控制方法。     

Investigation of plant extracts in traditional medicine of the Brazilian Cerrado against protozoans and yeasts

Aim of the study

To investigate the activities of the 217 plant extracts in traditional medicine of the Brazilian Cerrado against protozoans and yeasts.

Materials and methods

Plant extracts were prepared by the method of maceration using solvents of different polarities. The growth inhibition of chloroquine-resistant Plasmodium falciparum strain (FcB1) was determined by measuring the radioactivity of the tritiated hypoxanthine incorporated. Activity against Leishmania (Leishmania) chagasi and Trypanosoma cruzi was measured by the MTT colorimetric assay. The antifungal tests were carried out by using the CLSI method. The active extracts were tested also by cytotoxicity assay using NIH-3T3 cells of mammalian fibroblasts.

Results

Two hundred and seventeen extracts of plants were tested against Plasmodium falciparum. The eleven active extracts, belonging to eight plant species were evaluated against L. (L.) chagasi, Trypanosoma cruzi, yeasts and in NIH-3T3 cells. The results found in these biological models are consistent with the ethnopharmacological data of these plants. The ethyl acetate extract of Diospyros hispida root showed IC₅₀ values of 1 μg/mL against Plasmodium falciparum. This extract demonstrated no toxicity against mammalian cells, resulting in a significant selectivity index (SI) of 435.8. The dichloromethane extract of Calophyllum brasiliense root wood was active against Cryptococcus gattii LMGO 01 with MIC of 1.95 μg/mL; and Candida albicans ATCC 10231 and Candida krusei LMGO 174, both with MIC of 7.81 μg/mL. The same extract was also active against Plasmodium falciparum and L. (L.) chagasi with IC₅₀ of 6.7 and 27.6 μg/mL respectively. The ethyl acetate extract of Spiranthera odoratissima leaves was active against Cryptococcus gattii LMGO 01 with MIC of 31.25 μg/mL, and against Plasmodium falciparum with IC₅₀ of 9.2 μg/mL and Trypanosoma cruzi with IC₅₀ of 56.3 μg/mL.

Conclusion

The active extracts for protozoans and human pathogenic yeasts are considered promising to continue the search for the identification and development of leading compounds.     

Effect of feeding Mucuna pruriens on helminth parasite infestation in lambs

Ethnopharmacological relevance

Mucuna pruriens is a tropical legume anecdotally reputed to have anthelmintic properties. This study was conducted to examine the validity of such claims.

Aim of the study

The aim of this study was to determine if ingestion of Mucuna seeds reduces helminth parasite infestation in lambs.

Materials and methods

Thirty-six Dorper × Katahdin ram lambs were assigned to three treatments, a cottonseed meal based control diet, a diet in which Mucuna replaced cottonseed meal and the control diet with levamisole (7.5 mg/kg body weight) administration. All diets were isonitrogenous and isocaloric. The 12 lambs in each treatment were assigned randomly to 4 pens, each containing 3 lambs. Lambs were trickle infected three times per week by gavage with infectious Haemonchus contortus larvae (2000 larvae/lamb) for 3 weeks.

Results

Levamisole treatment decreased fecal egg counts by 87% and abomasal worm counts by 83%. Mucuna intake did not statistically affect fecal egg counts or abomasal worm counts, though numerical (P > 0.10) reductions of 7.4% and 18.1%, respectively were evident. Anemia indicators, feed intake, and lamb growth were unaffected by treatment.

Conclusions

Levamisole reduced the Haemonchus parasite burden in lambs significantly but feeding Mucuna reduced the burden by levels unlikely to eliminate the clinical effects of parasitism.     

Screening of the topical anti-inflammatory activity of the bark of Acacia cornigera Willdenow, Byrsonima crassifolia Kunth, Sweetia panamensis Yakovlev and the leaves of Sphagneticola trilobata Hitchcock

Ethnopharmacological relevance

An investigation of topical anti-inflammatory activity was undertaken on plants used in Central America traditional medicine.

Aim of study

Four herbal drugs used in the folk medicine of Central America to treat inflammatory skin affections (Acacia cornigera bark, Byrsonima crassifolia bark, Sphagneticola trilobata leaves and Sweetia panamensis bark) were evaluated for their topical anti-inflammatory activity.

Materials and methods

Petroleum ether, chloroform and methanol extracts were obtained for herbal medicines and then extracts were tested on Croton oil-induced ear dermatitis model in mice.

Results

Almost all the extracts reduced the Croton oil-induced ear dermatitis in mice and the chloroform ones showed the highest activity, with ID₅₀ (dose giving 50% oedema inhibition) values ranging from 112 μg/cm² (Byrsonima crassifolia) to 183 μg/cm² (Sphagneticola trilobata). As reference, ID₅₀ of the non-steroidal anti-inflammatory drug indomethacin was 93 μg/cm².

Conclusions

Lipophilic extracts from these species can be regarded as potential sources of anti-inflammatory principles.     

中国石斛属植物文献计量研究

石斛是珍稀濒危中药材,目前正处于快速发展阶段。为全面了解我国石斛属植物研究的历史和发展现状,作者以1954～2010年"中国知网中国学术期刊网络出版总库"收录的石斛研究文献为依据,采用文献计量学的原理和方法,对我国石斛属植物研究文献从文献年代分布、期刊分布与被引频率、主题分布、研究对象分布、作者与研究机构分布等方面进行了统计与分析。结果表明,我国石斛研究明显分为起步(2个)、停滞、平稳发展、快速上升5个阶段;期刊分布存在离散性与集中性并存的现象,已形成核心期刊研究群,并以《中国中药杂志》、《中草药》和《陕西中医》为代表;研究主题广泛涉及临床与药理、组织培养与种苗繁育、成分分析等多个领域,已经形成比较稳定的研究机构和团队,但研究对象差异显著,以铁皮石斛、金钗石斛和霍山石斛最为集中。我国石斛属植物的研究已取得显著成果,但种植产业发展缓慢,供需矛盾突出,预计种苗繁育与人工种植、产品开发、化学与药理等方面是未来的研究热点,其文献报道仍将进一步上升。     

厚朴与凹叶厚朴群体遗传学研究

目的:对厚朴与凹叶厚朴的群体遗传学进行研究,为中药厚朴的质量控制提供分子生药学方面的依据。方法:对厚朴与凹叶厚朴15个居群应用2个叶绿体基因间序列psbA-trnH和trnL-trnF进行PCR扩增并测序,计算厚朴与凹叶厚朴单倍型频率,用程序HaploNst分析遗传多样性和遗传结构,应用TCS version 1.13软件构建单倍型网状进化树。结果:厚朴与凹叶厚朴均无特有单倍型存在,但单倍型频率存在显著差异,已开始出现遗传分化的趋势,NST略大于GST。结论:厚朴与凹叶厚朴在遗传上已出现遗传分化的趋势,但尚未完全分化成彼此独立的单系。     

Experimental study on anti-inflammatory activity of a TCM recipe consisting of the supercritical fluid CO2 extract of Chrysanthemum indicum, Patchouli Oil and Zedoary Turmeric Oil in vivo

Ethnopharmacological relevance

Chrysanthemum indicum (Compositae) Linné, Pogostemon cablin (Blanco) Benth and Curcuma wenyujin (Zingiberaceae) Y. H. Chen et C. Ling are three of the extensively used herbal remedies among traditional Chinese medicines for the purpose of anti-inflammation. A traditional Chinese medicine (TCM) recipe named CPZ consisting extracts of the above three herbs, has shown noteworthy anti-influenza activity, which is closely related to its anti-inflammatory feature.

Aim of this study

To investigated the anti-inflammtory activity of CPZ in vivo for a further exploration of the recipe's anti-inflammatory properties.

Materials and methods

The anti-inflammatory property of CPZ on acute inflammation was evaluated by inflammatory models of dimethylbenzene (DMB)-induced ear vasodilatation and acetic acid-induced capillary permeability enhancement in mice, as well as the carrageenan-induced paw edema rat model, in which inflammation-related cytokine including prostaglandin E₂ (PGE₂), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and nitric oxide (NO) in the edematous paw tissue were determined by enzyme linked immunosorbent assay (ELISA). Moreover, effect of CPZ on chronic inflammation was observed through granuloma formation in rats subjected to cotton pellet implantation.

Results

CPZ (340, 170, and 85 mg/kg for mice, p.o.) not only decreased the DMB-induced ear vasodilatation but also attenuated capillary permeability under acetic acid challenge in mice. And the significant inhibition on carrageenan-induced paw edema was observed. Further more, the ELISA results showed that CPZ (170, 85, and 42.5 mg/kg for rats, p.o.) could up-regulate the level of IL-1β in the edema paw tissue of rats significantly while down-regulate that of PGE₂, but no apparent effect on TNF-α or NO was observed in the test. Besides, CPZ had a certain degree of restraining effect on the cotton pellet-induced granuloma formation in rats and the highest dose of 170 mg/kg even showed a significant suppression on it.

Conclusion

The above results indicated that CPZ possessed a potent anti-inflammatory activity, which is indicated to be closely associated with its regulation on IL-1β and PGE₂ thereby mediating the inflammatory response acting at an appropriate level.     

Regulation effects on abnormal glucose and lipid metabolism of TZQ-F,a new kind of Traditional Chinese Medicine

Aim of the study

Based on the recipe of the traditional anti-diabetic formula TZQ, we developed TZQ-F, a new formula including 8 fractions isolated from Red Paeony root, Mulberry leaf, Lotus leaf, Danshen root and Hawthorn leaf with a good quality assurance. The study was aimed at fraction preparation and effects of the fractions on abnormal glucose and lipid metabolism.

Materials and methods

The active fractions were obtained by macroporous resin, ion-exchange resin and polyamide resin column chromatographies. HPLC analyses were used for quality control. In vitro mechanism study included DPPH radical scavenging, AGEs formation inhibition, α-glucosidase inhibition and lipase inhibition, and rats on high-fat diet were used for in vivo study.

Results

In vitro mechanism study showed that among the 8 fractions, three of them had inhibition effects on intestinal disaccharase, three with inhibition effects on lipase, and five with effects of free radical scavenging. In vivo study showed that after 4 weeks of treatment, TZQ-F significantly decreased the levels of serum total cholesterol, TG, glucose, LDL-C and HDL-C in rats on high-fat diet. Consistent with the in vitro and in vivo results, histology study demonstrated that TZQ-F alleviated hepatic steatosis induced by high-fat diet.

Conclusions

TZQ-F possesses the potential regulation effects on abnormal glucose and lipid metabolism.     

Antifungal activity of Brazilian medicinal plants involved in popular treatment of mycoses

A survey of medicinal plants used to treat common mycoses was done in the Curituba district, Sergipe State, Brazil. One hundred inhabitants were interviewed by health agents and traditional healers. Four different plants were the most cited (more than 50% of the citations): Ziziphus joazeiro, Caesalpinia pyramidalis, Bumelia sartorum and Hymenea courbaril. The aqueous extracts obtained following traditional methods and using different parts of these plants, were submitted to drop agar diffusion tests for primary antimicrobial screening. Only the water infusion extract of Ziziphus joazeiro and Caesalpinea pyramidalis presented a significant antifungal activity against Trichophyton rubrum, Candida guilliermondii, Candida albicans, Cryptococcus neoformans and Fonsecaea pedrosoi, when compared to the antifungal agent amphotericin B. The minimal inhibitory concentration (MIC) of the bioactive extracts was evaluated by the microdilution method. Best activity with a MIC of 6.5 microg/ml for both extracts was observed against Trichophyton rubrum and Candida guilliermondii. Ziziphus joazeiro and Caesalpinea pyramidalis extracts presented also low acute toxicity in murine models. The present study validates the folk use of these plant extracts and indicates that they can be effective potential candidates for the development of new strategies to treat fungal infections.     

常用藏药“蒂达”(藏茵陈)基原物种药用合理性及资源利用价值评价

目的:对“蒂达”各品种不同基原植物药用合理性及其资源利用价值作出评价.方法:采用HPLC指纹图谱共有特征峰、结合SPSS聚类分析方法,评价“蒂达”不同品种基原植物的成分组成相似性.结果:“蒂达”不同品种涉及的来源有龙胆科獐牙菜属、花锚属、扁蕾属及肋柱花属共15种1变种33个样品,指纹图谱全谱相似度较差;以10个共有峰成分的相对面积聚类,其基原植物可分别聚为4个组;上述4个属的属间成分组成差异较大;其中獐牙菜属不同种间成分组成变化较大,而其他各属种间成分组成相似度较高.结论:上述龙胆科4属植物均含有具有保肝利胆生物活性的环烯醚萜类、(山)酮类及三萜类成分,藏医学将其归为“蒂达”一类药材使用具有一定的物质基础;鉴于“蒂达”类药材不同基原各品种间在成分组成上差异较大,分别制定各品种的HPLC指纹图谱,是鉴别和控制各品种质量的有效手段,可有效控制各基原品种质量.     

乌拉尔甘草HMGR基因cDNA的克隆与序列分析

目的:对乌拉尔甘草3.羟基-3-甲基戊二酰辅酶A还原酶(3-hydroxy-3methylglutary CoA reductase,HMGR)的cDNA克隆并进行序列分析.方法:根据NCBI数据库中的豆科其他物种HMGR的cDNA保守区设计引物,利用同源扩增和cDNA末端快速扩增技术从甘草根中获得目的基因;利用BLAST进行序列比对,ORF Finder寻找开发阅读框,Prosite分析蛋白质的基本结构域,Clustal x比对已有HMGR的氨基酸序列,并构建进化树.结果:得到1个全长为1 842 bp的HMGR的cDNA序列,含有1 722 bp的开放阅读框(open reading frame,ORF),编码573个氨基酸,具有HMGR家族的特异序列,推测的氨基酸序列与豌豆、蒺藜苜蓿的氨基酸序列一致性分别为84%,76%.结论:对甘草HMGR基因的cDNA进行了克隆,为进一步研究3-羟基-3-甲基戊二酰辅酶A在甘草酸生物合成途径中的作用提供了理论依据.