Identification and analysis of UGT genes associated with triterpenoid saponin in soapberry (Sapindus mukorossi Gaertn.).

BACKGROUND: Soapberry (Sapindus mukorossi) is an economically important multifunctional tree species. Triterpenoid saponins have many functions in soapberry. However, the types of uridine diphosphate (UDP) glucosyltransferases (UGTs) involved in the synthesis of triterpenoid saponins in soapberry have not been clarified. RESULTS: In this study, 42 SmUGTs were identified in soapberry, which were unevenly distributed on 12 chromosomes and had sequence lengths of 450 bp to 1638 bp, with an average of 1388 bp. The number of amino acids in SmUGTs was 149 to 545, with an average of 462. Most SmUGTs were acidic and hydrophilic unstable proteins, and their secondary structures were mainly α-helices and random coils. All had conserved UDPGT and PSPG-box domains. Phylogenetic analysis divided them into four subclasses, which glycosylated different carbon atoms. Prediction of cis-acting elements suggested roles of SmUGTs in plant development and responses to environmental stresses. The expression patterns of SmUGTs differed according to the developmental stage of fruits, as determined by transcriptomics and RT-qPCR. Co-expression network analysis of SmUGTs and related genes/transcription factors in the triterpenoid saponin synthesis pathway was also performed. The results indicated potential roles for many transcription factors, such as SmERFs, SmGATAs and SmMYBs. A correlation analysis showed that 42 SmUGTs were crucial in saponin synthesis in soapberry. CONCLUSIONS: Our findings suggest optimal targets for manipulating glycosylation in soapberry triterpenoid saponin biosynthesis; they also provide a theoretical foundation for further evaluation of the functions of SmUGTs and analyses of their biosynthetic mechanisms.

Regeneration of soapnut tree through somatic embryogenesis and assessment of genetic fidelity through ISSR and RAPD markers.

Somatic embryogenic system was developed in Sapindus mukorossi Gaertn. using rachis as explants from a mature tree. Explants showed callus initiation on Murashige and Skoog medium supplemented with TDZ (1-Phenyl-3-(1, 2, 3-thiadiazol-5-yl) urea), zeatin or 6-benzylaminopurine. Induction of somatic embryogenesis was achieved on both MS basal medium and MS medium supplemented with 8.88 µM 6-benzylaminopurine. Hundred percent embryogenesis was observed on MS medium supplemented with 8.88 µM 6-benzylaminopurine with maximum intensity of embryogenesis (51.92 ± 0.40 a). Maximum maturation of somatic embryos (92.86 ± 0.34 a) was observed on induction medium supplemented with 0.0378 µM abscisic and treated for 21 days. Germination of somatic embryos was maximum (77.33 ± 0.58 a) on MS medium supplemented with 8.88 µM 6-benzylaminopurine. In vitro raised plantlets were hardened, acclimatized and transferred to the field. Survival frequency of plantlets was 80 % in field conditions. The genetic fidelity of in vitro regenerated plants was also evaluated and compared with mother plant using random amplified polymorphic DNA and inter simple sequence repeat. Both markers showed similarity in molecular profile of mother plant and in vitro regenerated plants.

Protective effects of Sapindus mukorossi Gaertn against fatty liver disease induced by high fat diet in rats.

OBJECTIVES: Study the effects of alcohol extract of Sapindus mukorossi Gaertn (AESM) on the metabolism of blood fat, morphology of fenestrated liver sinusoidal endothelial cells (LSEC), and the ultrastructure of liver cells of the rats with non-alcoholic fatty liver disease (NAFLD). METHODS: Divide SD rats into control group, model group, simvastatin (7.2 mg/kg) group, and S.mukorossi Gaertn group with high dosage (0.5 g/kg), moderate dosage (0.1 g/kg), and low dosage (0.05 g/kg). After feeding with fat-rich nutrients for 3 weeks and establishing the model of hepatic adipose, conduct intragastric administration and provide the rats with fat-rich nutrients at the same time. At the 43rd day, take blood sample and measure aminotransferase and different indexes of blood fat; take hepatic tissue for pathological section, and observe the hepatic morphological patterns under light microscope; obtain and fix the hepatic tissue after injecting perfusate into the body, and observe the changes of fenestrated LSEC under scanning electron microscope; observe the ultrastructure of liver cells under transmission electron microscope. RESULTS: High-dosage alcohol extracts of S.mukorossi Gaertn can alleviate the AST, ALT, TC, TG, LDL, γ-GT, and ALP level, as well as raise the HDL and APN level in the serum of NAFLD-rat model. In addition, through the observation from light microscope and electron microscopes, the morphology of the hepatic tissue and liver cells as well as the recovery of the fenestrated LSEC in the treatment group has become normal. CONCLUSIONS: Alcohol extracts of S.mukorossi Gaertn can regulate the level of blood fat and improve the pathological changes of the hepatic tissues in NAFLD-rat model, which demonstrates the effects of down-regulating fat level and protecting liver.

Physicochemical and structural characterisation of protein isolate, globulin and albumin from soapnut seeds (Sapindus mukorossi Gaertn.).

The amino acid (AA) composition and physicochemical and conformational properties of protein isolate (SNPI), globulin (SNG) and albumin (SNA) fractions from soapnut seeds were evaluated. The essential AA of SNG, SNA and SNPI (except sulfur-containing AA) are sufficient for the FAO/WHO suggested requirements for 2-5year old infants. SNG and SNPI showed similar electrophoresis patterns and AA compositions, the subunit of those proteins consisted of two polypeptides linked by disulfide bonds. In contrast, SNA showed a different AA compositions and SDS-PAGE pattern. Both SNG and SNPI presented a typical U-shape protein solubility (PS)-pH profile, SNA showed a completely different PS-pH profile, especially at pH 2.0-4.0. The near-UV circular dichroism (CD), differential scanning calorimetry (DSC) and tryptophan fluorescence spectra analyses indicated that the flexibility in tertiary conformations decreased in the order: SNA>SNPI>SNG, while soapnut proteins had a similar secondary conformation, with a highly ordered structure (the ß-types), as evidenced by far-UV CD spectra.

Isolation of two sesquiterpene glycosides from Sapindus mukorossi Gaertn. with cytotoxic properties and analysis of their mechanism based on network pharmacology.

The anti-tumor effects of two compounds purified from Sapindus mukorossi Gaertn. (S. mukorossi.) on breast cancer in vitro were observed. Their chemical structures were identified as sesquiterpene glycosides, namely, Mukurozioside IIa and Mukurozioside IIb. The results of XTT assay indicated that their inhibition rates against three cancer cell lines (MCF-7, MDA-MB-231 and MDA-MB-435s) reached approximately 80% at a concentration of 200 µg/mL, which were higher than that of cyclophosphamide (below 40% at 200 µg/mL), and their 50% inhibiting concentrations were ranged from 120.73 to 154.01 µg/mL, indicating their inhibition were weaker than their parent fraction. Furthermore, the mechanism on breast cancer was predicted, and 22 targets including PTPN1, IL2 and VEGFA were relatively important. These results illustrated the anti-breast cancer activity of S. mukorossi was related to the two compounds with the structure of sesquiterpene glycosides, but they did not represent the full activity of their parent fraction.

The chemical profile and biological activity of different extracts of Sapindus mukorossi Gaertn. against Cutibacterium acnes.

This study evaluated the antibacterial activities of different extracts of Sapindus mukorossi Gaertn. (S. mukorossi) on Cutibacterium acnes (C. acnes). The extract solvent and procedure were screened, based on the yield of saponins and minimum inhibitory concentration (MIC). The results showed that the optimized product, fermentation and ethyl acetate extract by adding isoamyl alcohol from water extract of S. mukorossi (SWFEAI), had the highest yield of saponins (7.83 ± 0.26%) and the best antibacterial activity (MIC = 0.125 mg/mL) on C. acnes. The destroyed bacterial cell membrane and wall were observed by transmission electron microscopy, which then resulted in cell lysis and death. Furthermore, 20 compounds of SWFEAI were detected, among which oleanane-type triterpenoid saponins with molecular weights of 734, 750, 882, 924 and 966 were speculated to contribute to the antibacterial activities of SWFEAI. The results showed that SWFEAI could be a natural anti-acne agent.

Saponin fraction from Sapindus mukorossi Gaertn as a novel cosmetic additive: Extraction, biological evaluation, analysis of anti-acne mechanism and toxicity prediction.

ETHNOPHARMACOLOGICAL RELEVANCE: Sapindus mukorossi Gaertn. (S. mukorossi), known as 'mu huan zi' in Chinese folklore, belongs to the family Sapindaceae and it has been traditionally used for treating coughing and excessive salivation, removing freckle, whitening skin, etc. Evidence-based medicine also verified the antimicrobial, anti-tyrosinase and anti-acne activity of S. mukorossi extract, suggesting that it has the potential to be a pharmaceutical and cosmetic additive. AIM OF THE STUDY: The present study was intended to evaluate the freckle-removing and skin-whitening activities of S. mukorossi extracts, and further analyzing the potential anti-acne mechanism. METHODS: Saponin fractions were purified by using the semi-preparative high-performance liquid chromatography, and their antibacterial activity was detected against Propionibacterium acnes (P. acnes), which was the leading cause of inflamed lesions in acne vulgaris. The anti-lipase and anti-tyrosinase activities were assayed using a commercial kit, while the potential anti-acne mechanism was predicted on the basis of the network pharmacology. Active components of saponin fraction were identified by HPLC-MS analysis. Furthermore, the different toxicity level of compounds was predicted according to the quantitative structure-activity relationship, and the first application of crude extract and saponin fraction to facial masks was analyzed based on the comprehensive evaluation method. RESULTS: The saponin fraction (F4) purified from the fermentation liquid-based water extract (SWF) showed the best antibacterial activity against P. acnes ATCC 6919 with the MIC of 0.06 mg/mL, which was 33-fold of its parent SWF (with the MIC of 2.0 mg/mL). Compared with SWF, the application of F4 caused greater inhibition rates on lipase and tyrosinase. Chemical constituents of F4 were evaluated, from which four oleanane-type triterpenoid saponins were detected to contribute to the above biological activities of F4. The mechanism of the four compounds on anti-acne was predicted, and seven targets such as PTGS2 and F2RL1 were obtained to be important for the treatment of acne. The four compounds were also predicted to have different levels of toxicity to various species, and they were not harmful to rats. Besides, F4 and SWF were applied to facial masks and there was no significant influence on the physicochemical properties including pH, stability, and sensory characteristics. CONCLUSION: This work demonstrated that oleanane-type triterpenoid saponins were speculated to contribute to the skin-whitening, freckle-removing, and anti-acne activities of F4. These findings will facilitate the development of the S. mukorossi extract and the allied products as the new and natural anti-acne agent and cosmetic additives.

Human pulp tissue dissolution ability of different extracts of Sapindus mukorossi: An in vitro study.

Objective: Due to the many negative properties of sodium hypochlorite used in current root canal treatment, interest in biocompatible natural agents is increasing day by day. The aim of this study was to evaluate whether various extract solutions of Sapindus mukorossi have dissolution effects on human pulp tissues. Methods: Primarily powder extracts were obtained by extracting fruit shells of S. mukorossi in different solvents (ethanol, methanol, buthanol and distilled water). The test solutions were prepared and randomly separated into six groups with 10 samples in each group: ethanol extract, methanol extract, butanol extract, distilled water extract of S. mukorossi, sodium hypochlorite (NaOCl) and the control group. Among these, S. mukorossi extracts were separated into two subgroups, depending on their concentration level (50 µg/mL and 100 µg/mL). The pulp tissues of freshly extracted human molars were used for dissolution test. The weights of the pulpal tissues were measured and recorded for two times after the samples were placed in the solutions. Statistical analysis for all descriptive statistics was performed using SPSS 22 (P < 0.05). Results: Our results showed that maximum percent yield of preparation was obtained in methanol extract of S. mukorossi. Among all of the groups, the best dissolution capacity was seen in the NaOCl group (positive control group). Among S. mukorossi groups, the best tissue solvent solution was found in SMM group at 50 µg/mL and SMB group at 100 µg/mL. Conclusion: The different extracts of S. mukorossi had a capacity to dissolve pulp tissue but this capacity was less than NaOCl. Therefore, further studies will enable the creation of a commercial solution for clinical use by increasing the effectiveness of S. mukorossi while combining it with other endodontic irrigation solutions.

Human pulp tissue dissolution ability of different extracts of Sapindus mukorossi: An in vitro study / 中草药·英文版

Objective: Due to the many negative properties of sodium hypochlorite used in current root canal treatment, interest in biocompatible natural agents is increasing day by day. The aim of this study was to evaluate whether various extract solutions of Sapindus mukorossi have dissolution effects on human pulp tissues. Methods: Primarily powder extracts were obtained by extracting fruit shells of S. mukorossi in different solvents (ethanol, methanol, buthanol and distilled water). The test solutions were prepared and randomly separated into six groups with 10 samples in each group: ethanol extract, methanol extract, butanol extract, distilled water extract of S. mukorossi, sodium hypochlorite (NaOCl) and the control group. Among these, S. mukorossi extracts were separated into two subgroups, depending on their concentration level (50 µg/mL and 100 µg/mL). The pulp tissues of freshly extracted human molars were used for dissolution test. The weights of the pulpal tissues were measured and recorded for two times after the samples were placed in the solutions. Statistical analysis for all descriptive statistics was performed using SPSS 22 (P < 0.05). Results: Our results showed that maximum percent yield of preparation was obtained in methanol extract of S. mukorossi. Among all of the groups, the best dissolution capacity was seen in the NaOCl group (positive control group). Among S. mukorossi groups, the best tissue solvent solution was found in SMM group at 50 µg/mL and SMB group at 100 µg/mL. Conclusion: The different extracts of S. mukorossi had a capacity to dissolve pulp tissue but this capacity was less than NaOCl. Therefore, further studies will enable the creation of a commercial solution for clinical use by increasing the effectiveness of S. mukorossi while combining it with other endodontic irrigation solutions.

Purification of total saponins in Sapindus mukorossi by continuous foam separation / 中草药

Objective: The continuous foam separation was adopted to optimize the process conditions for separation of tatal saponins in Sapindus mukorossi. Methods: Orthogonal test and spectrophotometry were employed to determine the optimum conditions of foam separation using the enrichment ratio, yield, and purity of total saponins in S. mukorossi as evaluating indexes. Results: The best condition of the process was as follows: gas flow rate of 0.6 L/min, feed height of 40 cm, liquid flow rate of 35 mL/min with initial feed total saponins in S. mukorossi at concentration of 20 mg/L with pH 4.40. Conclusion: The yield and purity of the isolated total saponins in S. mukorossi were 23.64% and 90.27%, respectively. The continuous foam separation could conduct in relatively simple equipment, low energy consuming and provide the possibility to future industrial applications.