Purification and characterization of a novel antifungal protein from Bacillus subtilis strain B29"

An antifungal protein was isolated from a culture of Bacillus subtilis strain B29. The isolation procedure comprised ion exchange chromatography on diethylaminoethyl (DEAE)-52 cellulose and gel filtration chromatography on Bio-Gel P-100.The protein was absorbed on DEAE-cellulose and Bio-Gel P-100. The purified antifungal fraction was designated as B29I, with a molecular mass of 42.3 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), pl value 5.69 by isoelectric focusing (IEF)-PAGE, and 97.81% purity by high performance liquid chromatography (HPLC). B29I exhibited in-hibitory activity on mycelial growth in Fusarium oxysporum, Rhizoctonia solani, Fusarium moniliforme, and Sclerotinia scle-rotiorum. The 50% inhibitory concentrations (IC50) of its antifungal activity toward Fusarium oxysporum and Rhizoctonia solani were 45 and 112 μmol/L, respectively. B291 also demonstrated an inhibitory effect on conidial spore germination of Fusarium oxysporum and suppression of germ-tube elongation, and induced distortion, tumescence, and rupture of a portion of the germi-nated spores.     

Hydroxychavicol,a polyphenol from Piper betle leaf extract,induces cell cycle arrest and apoptosis in TP53-resistant HT-29 colon cancer cells

This study aims to elucidate the antiproliferative mechanism of hydroxychavicol(HC).Its effects on cell cycle,apoptosis,and the expression of c-Jun N-terminal kinase(JNK)and P38 mitogen-activated protein kinase(MAPK)in HT-29 colon cancer cells were investigated.HC was isolated from Piper betle leaf(PBL)and verified by high-performance liquid chromatography(HPLC),nuclear magnetic resonance(NMR),and gas chromatography-mass spectrometry(GC-MS).The cytotoxic effects of the standard drug 5-fluorouracil(5-FU),PBL water extract,and HC on HT-29 cells were measured after 24,48,and 72 h of treatment.Cell cycle and apoptosis modulation by 5-FU and HC treatments were investigated up to 30 h.Changes in phosphorylated JNK(pJNK)and P38(pP38)MAPK expression were observed up to 18 h.The half maximal inhibitory concentration(IC₅₀)values of HC(30μg/mL)and PBL water extract(380μg/mL)were achieved at 24 h,whereas the IC₅₀of 5-FU(50μmol/L)was obtained at 72 h.Cell cycle arrest at the G0/G1 phase in HC-treated cells was observed from12 h onwards.Higher apoptotic cell death in HC-treated cells compared to 5-FU-treated cells(P<0.05)was observed.High expression of pJNK and pP38 MAPK was observed at 12 h in HC-treated cells,but not in 5-FU-treated HT-29 cells(P<0.05).It is concluded that HC induces cell cycle arrest and apoptosis of HT-29 cells,with these actions possibly mediated by JNK and P38 MAPK.     

襄麦冬粗提物抗菌活性初步研究

以蒸馏水、甲醇、乙酸乙酯为溶剂,提取襄麦冬活性物质,并分别测定提取物对以下5种病原真菌：立枯丝核菌Rhizoctonia solani,灰葡萄孢菌Botrytis cinerea,齐整小核菌Sclerotium rolfsii,终极腐霉Pythium ultimum,尖镰孢菌黄瓜转化型Fusarium oxysporum f.sp cucumber菌丝生长和孢子萌发的抑制作用.结果表明襄麦冬的三种溶剂提取物对灰葡萄孢菌、终极腐霉、立枯丝核菌的菌丝生长均具有较强的抑制作用,而上述相同浓度的提取物对齐整小核菌、尖镰孢菌黄瓜转化型的生长未表现出明显的抑制效果.同时,三种溶剂提取物对立枯丝核菌、灰葡萄孢菌、齐整小核菌、终极腐霉、尖镰孢菌黄瓜转化型的孢子萌发均表现出一定的抑制作用.其中,襄麦冬乙酸乙酯提取物较蒸馏水和甲醇提取物对病原真菌的生长和孢子萌发的抑制效果明显.乙酸乙酯提取物相对于蒸馏水、甲醇提取物有较好的抑菌效果.襄麦冬提取物对灰葡萄孢菌、终极腐霉、立枯丝核菌的菌丝生长和立枯丝核菌、灰葡萄孢菌、齐整小核菌、终极腐霉、尖镰孢菌黄瓜转化型的孢子萌发有良好的抑制作用.     

Studies on the management of root-knot nematode, Meloidogyne incognita -wilt fungus, Fusarium oxysporum disease complex of green gram, Vigna radiata cv ML-1108

Studies were conducted under pot conditions to determine the comparative efficacy of carbofuran at 1 mg a.i./kg soil, bavistin at 1 mg a.i./kg soil, neem (Azadirachta indica) seed powder at 50 mg/kg soil, green mould (Trichoderma harzianum) at 50.0 ml/kg soil, rhizobacteria (Pseudomonas fluorescens) at 50.0 ml/kg soil against root-knot nematode,Meloidogyne incognita-wilt fungus,Fusarium oxysporum disease complex on green gram,Vigna radiata cv ML-1108. All the treatments significantly improved the growth of the plants as compared to untreated inoculated plants. Analysis of data showed that carbofuran andA. indica seed powder increased plant growth and yield significantly more in comparison to bavistin andP. fluorescens. Carbofuran was highly effective against nematode, bavistin against fungus,A. indica seed powder against both the pathogens and both the bioagents were moderately effective against both the pathogens.     

Purification and some properties of a β-glucanase from a strain,Trichoderma reesei GXC

β-glucanase was purified from a solid-state culture ofTrichoderma reesei on wheat bran in three steps which comprised ammonium sulfate precipitation, Sephadex G-100 chromatography, and DEAE-Sephadex A-50 chromatography. The molecular mass was determined to be 35.21 kilodaltons by sodium dodecyl sulfate-12.5% polyacrylamide gel electrophoresis. The β-glucanase at low pHs was more stable than that at high pHs, and optimum pH was 5.0. The optimum temperature was 60°C, and β-glucanase was relatively stable at below 40°C for 60 min. TheK _m of the enzyme on β-glucan was 10.86 mg/ml, and theV _max on β-glucan was 14286 μmol of glucose equivalents per mg of the pure enzyme per min. The β-glucanase activity was significantly inhibited by Fe³⁺ ions, and was reduced in the presence of Cu²⁺ ions, Mn²⁺ ions and Mg²⁺ ions at 5 mmol/L and 10 mmol/L, respectively. The β-glucanase activity was stimulated by Co²⁺ ions, Ca²⁺ ions, Zn²⁺ ions, and Fe²⁺ ions at 1 mmol/L and 5 mmol/L, respectively. Project supported by Foundation for University Key Teacher of the State Ministry of Education, the National Natural Science Foundation of China (No. 30000118) and Zhejiang Provincial Natural Science Foundation of China (No. 399409).     

Purification and studies on characteristics of cholinesterases from Daphnia magna

Due to their significant value in both economy and ecology, Daphnia had long been employed to investigate in vivo response of cholinesterase (ChE) in anticholinesterase exposures, whereas the type constitution and property of the enzyme remained unclear. A type of ChE was purified from Daphnia magna using a three-step procedure, i.e., Triton X-100 extraction, ammonium sulfate precipitation, and diethylaminoethyl (DEAE)-Sepharose?-Fast-Flow chromatography. According to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), molecular mass of the purified ChE was estimated to be 84 kDa. Based on substrate studies, the purified enzyme preferred butyrylthiocholine iodide (BTCh) [with maximum velocity (V _max)/Michaelis constant (K _m)=8.428 L/(min·mg protein)] to acetylthiocholine iodide (ATCh) [with V _max/K _m=5.346 L/(min·mg protein)] as its substrate. Activity of the purified enzyme was suppressed by high concentrations of either ATCh or BTCh. Inhibitor studies showed that the purified enzyme was more sensitive towards inhibition by tetraisopropylpyrophosphoramide (iso-OMPA) than by 1,5-bis(4-allyldimethylammoniumphenyl) pentan-3-one dibromide (BW284C51). Result of the study suggested that the purified ChE was more like a type of pseudocholinesterase, and it also suggested that Daphnia magna contained multiple types of ChE in their bodies.     

Purification and relationship with gastric disease of a 130 kDa (CagA) protein of<Emphasis Type="Italic">Helicobacter pylori</Emphasis>

Objective: The aims of this research were to purify and identify the 130 kDa (CagA) protein ofH. pylori clinical isolate HP97002 and evaluate the relationships between the purified 130 kDa (CagA) protein and gastric diseases. Methods: The procedure for isolating the protein included 6 mol/L guanidine extract, size exclusion chromatography and elusion from gel. Sera of 68 patients with gastric diseases (44 with chronic gastritis, 15 with atrophic gastritis, 7 with peptic ulcer disease, 2 with gastric cancer) were obtained, and the serological response to CagA was studied by Western-blot using the purified protein. Results: The purified protein was 130 kDa and preserved good antigenicity and revealed basic isoelectric point about of 8.1. Among 68 sera, 43 sera could recognize the purified protein associated with chronic gastritis 47.7% (21/44), atrophic gastritis 86.7% (13/15), peptic ulcer disease 100% (7/7), gastric cancer 100% (2/2). Compared with each other, the difference was significant (χ²=13.327,P=0.004), and 130 kDa (CagA) protein was associated with severe gastric diseases (r _s=0.442,P=0.001). Conclusion: The 130 kDa (CagA) protein was associated with severe gastric diseases. Project supported by the China Medical Board (96-628) and Zhejiang Province Hygiene Bureau (2000 A055)     

Cloning and efficient expression of <Emphasis Type="Italic">Bacillus sp.</Emphasis> BH072 <Emphasis Type="Italic">tasA</Emphasis> gene in <Emphasis Type="Italic">Escherichia coli</Emphasis>

The Bacillus strain BH072 isolated from a honey sample showed strong antifungal activity against phytopathogen. Gene cloning test demonstrated that the strain had a tasA gene encoding an antifungal TasA protein. Although the wild strain simultaneously produced various antifungal substances, only the physicochemical property and antifungal activity of TasA protein were unclear due to the difficulty in extraction. In this study, tasA gene encoding the protein from Bacillus sp. BH072 was amplified by using the polymerase chain reaction (PCR) method and cloned into pET 28a (+) vector, and then expressed in host cells Escherichia coli BL21 (DE3). The expressed proteins were collected by centrifugation and ultrasonic treatment, and then purified by using nickel-nitrilotriacetic acid (Ni-NTA) metal affinity column and dialysis methods. The result of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) test showed that an expected protein band appeared with a size of 31 kDa. The expressed products possessed antifungal activity against the phytopathogenic indicator strain Botrytis cinerea. A genetically engineered strain tasA of E. coli was established in this study which can efficiently express Tas A protein.     

Effects of total dissolved gas supersaturated water on lethality and catalase activity of Chinese sucker (Myxocyprinus asiaticus Bleeker)

Total dissolved gas (TDG) supersaturation caused by dam sluicing can result in gas bubble trauma (GBT) in fish and threaten their survival. In the present study, Chinese suckers (Myxocyprinus asiaticus Bleeker) were exposed to TDG supersaturated water at levels ranging from 120% to 145% for 48 h. The median lethal concentration (LC₅₀) and the median lethal time (LT₅₀) were determined to evaluate acute lethal effects on Chinese suckers. The results showed that the LC₅₀ values of 4, 6, 8, and 10 h were 142%, 137%, 135%, and 130%, respectively. The LT₅₀ values were 3.2, 4.7, 7.8, 9.2, and 43.4 h, respectively, when TDG supersaturated levels were 145%, 140%, 135%, 130%, and 125%. Furthermore, the biological responses in Chinese suckers were studied by assaying the catalase (CAT) activities in gills and muscles at the supersaturation level of 140% within LT₅₀. The CAT activities in the gills and muscle tissues exhibited a regularity of a decrease after an increase. CAT activities in the muscles were increased significantly at 3/5LT₅₀ (P<0.05) and then came back to the normal level. However, there were no significant differences between the treatment group (TDG level of 140%) and the control group (TDG level of 100%) on CAT activities in the gills before 3/5LT50 (P>0.05), but the activities were significantly lower than the normal level at 4/5LT₅₀ and LT₅₀ (P<0.05).     

Separation of mandelic acid and its derivatives with new immobilized cellulose chiral stationary phase

A new liquid chromatographic method has been developed for the chiral separation of the enantiomers of mandelic acid and their derivatives 2-chloromandelic acid, 4-hydroxymandelic acid, 4-methoxymandelic acid, and 3,4,5-trismethoxymandelic acid. The enantiomers were separated by a CHIRALPAK® IC (250 mm×4.6 mm, 5 μm). Mandelic acid, 4-methoxymandelic acid, and 3,4,5-trismethoxymandelic acid were baseline resolved (resolution factor (R _S)=2.21, R _S=2.14, and R _S=3.70, respectively). In contrast, the enantioselectivities between CHIRALPAK® IC and 2-chloromandelic acid and 4-hydroxymandelic acid investigated were low. By comparing the chromatographs of mandelic acid enantiomers and mandelic acid spiked with (R)-mandelic acid, it was determined that the first effluent was (R)-mandelic acid.     

萎叶提取物增强5-氟尿嘧啶对结肠癌细胞HT29和HCT116的生长抑制作用

研究目的：探讨萎叶（PB）提取物对5-氟尿嘧啶（5-FU）抑制结肠癌细胞HT29和HCT116生长的影响。研究方法：HT29和HCT116细胞分别给予PB、5-FU以及两种药物联合治疗24小时,应用等效线图法分析PB和5-FU的药效学相互作用,AnnexinV／PI染色法检测HT29和HCT116细胞的凋丁L=情况,高效液相色谱法排除PB和5-FU间任何可能的相互化学作用。重要结论：联合PB,低剂量5-FU可以在短时间内起到细胞毒作用,而单独应用PB或5-FU治疗较联合治疗可以诱导更多细胞发生凋亡。进一步采用等效线图法分析显示PB和5-Fu的联合作用在抑制结肠癌细胞HT29和HCT116的生长中分别体现出协同和拮抗作用。因此可以认为在HT29细胞中,PB使得较低剂量5-FU发挥最大抑制结肠癌细胞生长效果,然而在HCT116细胞中,PB没有显著降低5-FU的药物浓度,说明PB和5-FU的相互作用不仅仅体现在诱导细胞凋亡方面。     

Effects of levobupivacaine and bupivacaine on rat myometrium

Objective： To study the effect of levobupivacaine and bupivacaine on the contractility of isolated uterine muscle strips from pregnant and non-pregnant female rats. Methods： Full-thick myometrial strips were prepared from 18- to 2 l-day pregnant （n=8） and non-pregnant rats （n=7）. After contractions became regular, strips were exposed to cumulative concentrations of the two drugs from 10^-8 to 10^-4 mol/L, amplitude and frequency of the uterine contraction was recorded. Results： Two local anesthetics caused a concentration dependent inhibition on contractility of myometrial strips from pregnant and non-pregnant rats. In the myometrium from non-pregnant rats, -log/C50 of levobupivacaine and bupivacaine were 4.85 and 4.25 respectively. In the myometrium from pregnant rats, similar concentrations of levobupivacaine and bupivacaine were observed, -log/C50 were 2.7 and 2.9 respectively. Levobupivacaine produced an increase in amplitude of contractions, while bupivacaine showed an increased trend in frequency. Conclusion： These results demonstrate that levobupivacaine and bupivacaine may inhibit myometrium contractility. The inhibitory effect of levobupivacaine or bupivacaine is not enhanced by gestation in rat. Levobupivacaine may have more positive influence than bupivacaine in pregnant myometrium.     

Inhibitory mechanism of angiotensin-converting enzyme inhibitory peptides from black tea

The aim of this work is to discover the inhibitory mechanism of tea peptides and to analyse the affinities between the peptides and the angiotensin-converting enzyme(ACE) as well as the stability of the complexes using in vitro and in silico methods. Four peptide sequences identified from tea, namely peptides I, II, III, and IV, were used to examine ACE inhibition and kinetics. The half maximal inhibitory concentration(IC_(50)) values of the four peptides were(210.03±18.29),(178.91±5.18),(196.31±2.87), and(121.11±3.38) μmol/L, respectively. The results of Lineweaver-Burk plots showed that peptides I, II, and IV inhibited ACE activity in an uncompetitive manner, which requires the presence of substrate. Peptide III inhibited ACE in a noncompetitive manner, for which the presence of substrate is not necessary. The docking simulations showed that the four peptides did not bind to the active sites of ACE, indicating that the four peptides are allosteric inhibitors. The binding free energies calculated from molecular dynamic(MD) simulation were-72.47,-42.20,-52.10, and-67.14 kcal/mol(1 kcal=4.186 kJ),r espectively. The lower IC_(50) value of peptide IV may be attributed to its stability when docking with ACE and changes in the flexibility and unfolding of ACE. These four bioactive peptides with ACE inhibitory ability can be incorporated into novel functional ingredients of black tea.     

Ultrasound extraction optimization,structural features,and antioxidant activity of polysaccharides from <Emphasis Type="Italic">Tricholoma matsutake</Emphasis>

An ultrasonic-assisted technique was employed to extract crude polysaccharide from Tricholoma matsutake fruiting bodies. Single-factor tests and orthogonal experimental design (L ₉(3³)) were used to obtain the optimal extraction conditions. Results showed that the optimal parameters were as follows: ultrasonic temperature, 40 °C; ultrasonic time, 50 min; water to raw material ratio, 25 ml/g; ultrasonic frequency, 45 kHz; and ultrasonic power, 100 W. Three novel T. matsutake polysaccharide (TMP) fractions (TMP30, TMP60, and TMP80) were isolated and purified from TMP by stepwise alcohol precipitation. Their preliminary structural features were determined by high-performance anion-exchange chromatography with pulsed-amperometric detection (HPAEC-PAD) and Fourier transform infrared spectrophotometer (FT-IR) analyses. Furthermore, their in vitro antioxidant activity was investigated in terms of a reducing power assay and the scavenging rates of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radicals. The order of the various fractions based on their antioxidant activity was TMP80>TMP>TMP60>TMP30. These findings suggested that novel polysaccharide fractions from T. matsutake, especially TMP80, could be promising active macromolecules for biomedical use.     

Microcalorimetry studies of the antimicrobial actions of Aconitum alkaloids

The metabolic activity of organisms can be measured by recording the heat output using microcalorimetry. In this paper, the total alkaloids in the traditional Chinese medicine Radix Aconiti Lateralis were extracted and applied to Escherichia coli and Staphylococcus aureus. The effect of alkaloids on bacteria growth was studied by microcalorimetry. The power-time curves were plotted with a thermal activity monitor (TAM) air isothermal microcalorimeter and parameters such as growth rate constant (μ), peak-time (T_m), inhibitory ratio (I), and enhancement ratio (E) were calculated. The relationships between the concentration of Aconitum alkaloids and μ of E. coli or S. aureus were discussed. The results showed that Aconitum alkaloids had little effect on E. coli and had a potentially inhibitory effect on the growth of S. aureus.     

Antimelanogenic effects of <Emphasis Type="Italic">Inula britannica</Emphasis> flower petal extract fermented by <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis> KCCM 11613P

The inhibitory effects of Lactobacillus plantarum-fermented and non-fermented Inula britannica extracts on the tyrosinase activity were comparatively investigated to examine whether and how they improve the whitening activity, and the contents of total flavonoids and polyphenolics as bioactive compounds were determined. The skin whitening activity using in vitro or ex vivo tyrosinase and L-3,4-dihydroxyphenylalanine (L-DOPA) staining was examined. The total flavonoid content (TFC) was increased by 13.4% after 72 h-fermentation. The viabilities of the B16F10 cells treated with the fermented and non-fermented control extracts were 100.26% and 92.15% at 500 μg/ml, respectively. In addition, the inhibition of tyrosinase activity was increased by the fermented samples from 29.33% to 41.74% following fermentation for up to 72 h. The tyrosinase activity of the untreated control group was increased to 145.69% in B16F10 cells. The results showed that I. britannica fermented by L. plantarum dose-dependently inhibited tyrosinase activity, which was stimulated by α-melanocyte stimulating hormone. These results suggest that lactic fermented I. britannica extracts can be used as effective skin-whitening materials.     

一株活性细菌的抑菌活性研究

以实验中发现的一株细菌（ZXK）为材料,通过摇瓶发酵的方法获得其发酵液,采用对峙培养法和生长速率法分别测定了菌体和发酵液对棉花黄萎病菌Verticillium dahliae,小麦赤霉病菌病菌Gibberella zeae,水稻纹枯病菌Thanatephorus cucumeris,苹果炭疽病菌Glomere Uacingulata小麦纹枯病菌Rhizoctonia cerealis,番茄灰霉病菌Botrytis cinerea,辣椒疫霉病菌Phytophthora capsici等7种植物病原真菌的生物活性．结果表明：ZXK菌体及其发酵液对7种病原菌都有较好的抑制作用,ZXK发酵液能够完全抑制苹果炭疽病菌、水稻纹枯病菌、小麦赤霉病菌和小麦纹枯病菌菌丝的生长,对棉花黄萎病菌和番茄灰霉病菌的抑制率也达到了90％以上;发酵液经离心、过滤和高压蒸汽灭菌处理后仍具有生物活性,其中以高压蒸汽灭菌处理的发酵液样品的抑菌活性损失最大．     

Antioxidant activity and protective effect of Clitoria ternatea flower extract on testicular damage induced by ketoconazole in rats

Background

Ketoconazole (KET), an antifungal drug, has adverse effects on the male reproductive system. Pre-treatments with antioxidant plant against testicular damage induced by KET are required. The flowers of Clitoria ternatea (CT) are proven to have hepatoprotective potential. However, the protective effect on KET-induced testicular damage has not been reported.

Objective

To investigate the protective effect of CT flower extracts with antioxidant activity on male reproductive parameters including sperm concentration, serum testosterone level, histopathology of the testis, and testicular tyrosine phosphorylation levels in rats induced with KET.

Methods

The antioxidant activity of CT flower extracts was determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays. Male rats were treated with CT flower extracts (10, 50, or 100 mg/kg BW) or distilled water via a gastric tube for 28 d (preventive period: Days 1–21) and induced by KET (100 mg/kg BW) via intraperitoneal injection for 7 d (induction period: Days 22–28). After the experiment, all animals were examined for the weights of the testis, epididymis plus vas deferens and seminal vesicle, serum testosterone levels, sperm concentration, histological structures and diameter of testis, and testicular tyrosine phosphorylation levels by immunoblotting.

Results

The CT flower extracts had capabilities for DPPH scavenging and high reducing power. At 100 mg/kg BW, the extract had no toxic effects on the male reproductive system. Significantly, in CT+KET groups, CT flower extracts (50 and 100 mg/kg BW) alleviated the reduction of reproductive organ weight parameters, testosterone levels, and sperm concentration. In addition, CT flower extracts gave protection from testicular damage in KET-induced rats. Moreover, in the CT100+KET group, CT flower extracts significantly enhanced the expression of a testicular 50-kDa tyrosine phosphorylated protein compared with that of other groups.

Conclusions

C. ternatea flower extracts possessing antioxidant activity are not harmful to the male reproductive system and can protect against testicular damage in KET-induced rats.     

Cytotoxicity and enhancement activity of essential oil from Zanthoxylum bungeanum Maxim. as a natural transdermal penetration enhancer简

The aim of this present study is to investigate the effect of Zanthoxylum bungeanum oil (essential oil from Z. bungeanum Maxim.) on cytotoxicity and the transdermal permeation of 5-fluorouracil and indomethacin. The cytotoxicity of Z. bungeanum oil on dermal fibroblasts and epidermal keratinocytes was studied using an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The rat skin was employed to determine the percutaneous penetration enhancement effect of Z. bungeanum oil on hydrophilic and lipophilic model drugs, i.e., 5-fluorouracil and indomethacin. The secondary structure changes of the rat stratum corneum (SC) were determined using attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), and saturated solubilities and SC/vehicle partition coefficients of two model drugs with and without Z. bungeanum oil were also measured to understand its related mechanisms of action. It was found that the half maximal inhibitory concentration (IC₅₀) values of Z. bungeanum oil were significantly lower in HaCaT and CCC-ESF-1 cell lines compared to the well-established and standard penetration enhancer Azone. The Z. bungeanum oil at various concentrations effectively facilitated the percutaneous penetration of two model drugs across the rat skin. In addition, the mechanisms of permeation enhancement by Z. bungeanum oil could be explained with saturated solubility, SC/vehicle partition coefficient, and secondary structure changes of SC.