Mechanism of Bazi Bushen capsule in delaying the senescence of mesenchymal stem cells based on network pharmacology and experimental validation.

Ageing is becoming an increasingly serious problem; therefore, there is an urgent need to find safe and effective anti-ageing drugs. Aims: To investigate the effects of Bazi Bushen capsule (BZBS) on the senescence of mesenchymal stem cells (MSCs) and explore its mechanism of action. Methods: Network pharmacology was used to predict the targets of BZBS in delaying senescence in MSCs. For in vitro studies, MSCs were treated with D-gal, BZBS, and NMN, and cell viability, cell senescence, stemness-related genes, and cell cycle were studied using cell counting kit-8 (CCK-8) assay, SA-ß-galactosidase (SA-ß-gal) staining, Quantitative Real-Time PCR (qPCR) and flow cytometry (FCM), respectively. Alkaline phosphatase (ALP), alizarin red, and oil red staining were used to determine the osteogenic and lipid differentiation abilities of MSCs. Finally, the expression of senescence-related genes and cyclin-related factors was detected by qPCR and western blotting. Results: Network pharmacological analysis suggested that BZBS delayed cell senescence by interfering in the cell cycle. Our in vitro studies suggested that BZBS could significantly increase cell viability (P < 0.01), decrease the quantity of ß-galactosidase+ cells (P < 0.01), downregulate p16 and p21 (P < 0.05, P < 0.01), improve adipogenic and osteogenic differentiation, and upregulate Nanog, OCT4 and SOX2 genes (P < 0.05, P < 0.01) in senescent MSCs. Moreover, BZBS significantly reduced the proportion of senescent MSCs in the G0/G1 phase (P < 0.01) and enhanced the expression of CDK4, Cyclin D1, and E2F1 (P < 0.05, P < 0.01, respectively). Upon treatment with HY-50767A, a CDK4 inhibitor, the upregulation of E2F1 was no longer observed in the BZBS group. Conclusions: BZBS can protect MSCs against D-gal-induced senescence, which may be associated with cell cycle regulation via the Cyclin D1/CDK4/E2F1 signalling pathway.

Qili Qiangxin (QLQX) capsule as a multi-functional traditional Chinese medicine in treating chronic heart failure (CHF): A review of ingredients, molecular, cellular, and pharmacological mechanisms.

Chronic heart failure (CHF) is a key part of cardiovascular continuum. Under the guidance of the theory of vessel-collateral doctrine, the present study proposes therapeutic benefits of Qili Qiangxin (QLQX) capsules, an innovative Chinese medicine, on chronic heart failure. The studies show that multiple targets of the drug on CHF, including enhancing myocardial systole, promoting urine excretion, inhibiting excessive activation of the neuroendocrine system, preventing ventricular remodeling by inhibiting inflammatory response, myocardial fibrosis, apoptosis and autophagy, enhancing myocardial energy metabolism, promoting angiogenesis, and improving endothelial function. Investigation on the effects and mechanism of the drug is beneficial to the treatment of chronic heart failure (CHF) through multiple targets and/or signaling pathways. Meanwhile, it provides new insights to further understand other refractory diseases in the cardiovascular continuum, and it also has an important theoretical and practical significance in enhancing prevention and therapeutic effect of traditional Chinese medicine for these diseases.

Repositioning of clinically approved drug Bazi Bushen capsule for treatment of Aizheimer's disease using network pharmacology approach and in vitro experimental validation.

Aims: To explore the new indications and key mechanism of Bazi Bushen capsule (BZBS) by network pharmacology and in vitro experiment. Methods: The ingredients library of BZBS was constructed by retrieving multiple TCM databases. The potential target profiles of the components were predicted by target prediction algorithms based on different principles, and validated by using known activity data. The target spectrum of BZBS with high reliability was screened by considering the source of the targets and the node degree in compound-target (C-T) network. Subsequently, new indications for BZBS were predicted by disease ontology (DO) enrichment analysis and initially validated by GO and KEGG pathway enrichment analysis. Furthermore, the target sets of BZBS acting on AD signaling pathway were identified by intersection analysis. Based on STRING database, the PPI network of target was constructed and their node degree was calculated. Two Alzheimer's disease (AD) cell models, BV-2 and SH-SY5Y, were used to preliminarily verify the anti-AD efficacy and mechanism of BZBS in vitro. Results: In total, 1499 non-repeated ingredients were obtained from 16 herbs in BZBS formula, and 1320 BZBS targets with high confidence were predicted. Disease enrichment results strongly suggested that BZBS formula has the potential to be used in the treatment of AD. GO and KEGG enrichment results provide a preliminary verification of this point. Among them, 113 functional targets of BZBS belong to AD pathway. A PPI network containing 113 functional targets and 1051 edges for the treatment of AD was constructed. In vitro experiments showed that BZBS could significantly reduce the release of TNF-α and IL-6 and the expression of COX-2 and PSEN1 in Aß25-35-induced BV-2 cells, which may be related to the regulation of ERK1/2/NF-κB signaling pathway. BZBS reduced the apoptosis rate of Aß25-35 induced SH-SY5Y cells, significantly increased mitochondrial membrane potential, reduced the expression of Caspase3 active fragment and PSEN1, and increased the expression of IDE. This may be related to the regulation of GSK-3ß/ß-catenin signaling pathway. Conclusions: BZBS formula has a potential use in the treatment of AD, which is achieved through regulation of ERK1/2, NF-κB signaling pathways, and GSK-3ß/ß-catenin signaling pathway. Furthermore, the network pharmacology technology is a feasible drug repurposing strategy to reposition new clinical use of approved TCM and explore the mechanism of action. The study lays a foundation for the subsequent in-depth study of BZBS in the treatment of AD and provides a basis for its application in the clinical treatment of AD.

Complete Mitochondrial Genome of Piophila casei (Diptera: Piophilidae): Genome Description and Phylogenetic Implications.

Piophila casei is a flesh-feeding Diptera insect that adversely affects foodstuffs, such as dry-cured ham and cheese, and decaying human and animal carcasses. However, the unknown mitochondrial genome of P. casei can provide information on its genetic structure and phylogenetic position, which is of great significance to the research on its prevention and control. Therefore, we sequenced, annotated, and analyzed the previously unknown complete mitochondrial genome of P. casei. The complete mt genome of P. casei is a typical circular DNA, 15,785 bp in length, with a high A + T content of 76.6%. It contains 13 protein-coding genes (PCG), 2 ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and 1 control region. Phylogenetic analysis of 25 Diptera species was conducted using Bayesian and maximum likelihood methods, and their divergence times were inferred. The comparison of the mt genomes from two morphologically similar insects P. casei and Piophila megastigmata indicates a divergence time of 7.28 MYA between these species. The study provides a reference for understanding the forensic medicine, taxonomy, and genetics of P. casei.

A Long-Read Genome Assembly of a Native Mite in China Pyemotes zhonghuajia Yu, Zhang & He (Prostigmata: Pyemotidae) Reveals Gene Expansion in Toxin-Related Gene Families.

Pyemotes zhonghuajia Yu, Zhang & He (Prostigmata: Pyemotidae), discovered in China, has been demonstrated as a high-efficient natural enemy in controlling many agricultural and forestry pests. This mite injects toxins into the host (eggs, larvae, pupae, and adults), resulting in its paralyzation and then gets nourishment for reproductive development. These toxins have been approved to be mammal-safe, which have the potential to be used as biocontrol pesticides. Toxin proteins have been identified from many insects, especially those from the orders Scorpions and Araneae, some of which are now widely used as efficient biocontrol pesticides. However, toxin proteins in mites are not yet understood. In this study, we assembled the genome of P. zhonghuajia using PacBio technology and then identified toxin-related genes that are likely to be responsible for the paralytic process of P. zhonghuajia. The genome assembly has a size of 71.943 Mb, including 20 contigs with a N50 length of 21.248 Mb and a BUSCO completeness ratio of 90.6% (n = 1367). These contigs were subsequently assigned to three chromosomes. There were 11,183 protein coding genes annotated, which were assessed with 91.2% BUSCO completeness (n = 1066). Neurotoxin and dermonecrotic toxin gene families were significantly expanded within the genus of Pyemotes and they also formed several gene clusters on the chromosomes. Most of the genes from these two families and all of the three agatoxin genes were shown with higher expression in the one-day-old mites compared to the seven-day-pregnant mites, supporting that the one-day-old mites cause paralyzation and even death of the host. The identification of these toxin proteins may provide insights into how to improve the parasitism efficiency of this mite, and the purification of these proteins may be used to develop new biological pesticides.

A chromosome-level genome assembly of Neotoxoptera formosana (Takahashi, 1921) (Hemiptera: Aphididae).

Neotoxoptera formosana (Takahashi), the onion aphid, is an oligophagous pest that mainly feeds on plants from the Allium genus. It sucks nutrients from the plants and indirectly acts as a vector for plant viruses. This aphid causes severe economic losses to Allium tuberosum agriculture in China. To better understand the host plant specificity of N. formosana on Allium plants and provide essential information for the control of this pest, we generated the entire genome using Pacific Biosciences long-read sequencing and Hi-C data. Six chromosomes were assembled to give a final size of 372.470 Mb, with an N50 scaffold of 66.911 Mb. The final draft genome assembly, from 192 Gb of raw data, was approximately 371.791 Mb in size, with an N50 contig of 24.99 Kb and an N50 scaffold of 2.637 Mb. The average GC content was 30.96%. We identified 73 Mb (31.22%) of repetitive sequences, 14,175 protein-coding genes, and 719 noncoding RNAs. The phylogenetic analysis showed that N. formosana and Pentalonia nigronervosa are sister groups. We found significantly expanded gene families that were involved in the THAP domain, the DDE superfamily endonuclease, zinc finger, immunity (ankyrin repeats), digestive enzyme (serine carboxypeptidase) and chemosensory receptor. This genome assembly could provide a solid foundation for future studies on the host specificity of N. formosana and pesticide-resistant aphid management.

A Chromosome-Level Genome Assembly of the Parasitic Wasp Chelonus formosanus Sonan 1932 (Hymenoptera: Braconidae).

Chelonus formosanus Sonan 1932 (Hymenoptera: Braconidae) is a wasp capable of parasitizing a variety of lepidopteran pests at the "egg-larval" stage which distributes throughout Taiwan, Guangdong, Zhejiang, and Hainan provinces of China. This wasp has been successfully used to control pests such as Spodoptera litura Fabricius, 1775, Spodoptera frugiperda (JE Smith, 1797), Spodoptera exigua (Hübner, 1808), and Helicoverpa armigera (Hübner, 1808). So far, there is only one genome assembled from the Chelonus genus [Chelonus insularis (Cresson, 1865)] and it is fragmented with 455 scaffolds. Here, we report a chromosome-level genome assembly of C. formosanus, which was sequenced using PacBio, Illumina, and Hi-C technologies. The long reads were 35.4 Gb (∼150× coverage) with an average length of 15.23 kb. The size of the genome assembly was 139.59 Mb. More than 99.46% of the assembled sequences were anchored to seven pseudochromosomes (138.84 Mb). The Benchmarking University Single-Copy Orthologs (BUSCO) assessment results showed 99.0% of the 1,367 genes (insect_odb10 database) were completely present. We annotated 11,242 protein-coding genes including 98.6% of BUSCO complete genes that were recovered. Nearly one-fourth of the genome assembly (22.25%) was annotated as repetitive sequences and 324 noncoding RNAs were predicted. There were 58 gene families found with significant expansion including allelopathic families (odorant receptors and ionotropic receptors), which may play a crucial role in efficiently locating a wide range of hosts. This high-quality genome assembly and annotation could provide a highly valuable resource of parasitic wasp for the biological control of Lepidoptera pest.

A novel colorimetric aptasensor for sensitive tetracycline detection based on the peroxidase-like activity of Fe3O4@Cu nanoparticles and "sandwich" oligonucleotide hybridization.

A novel colorimetric aptasensor has been developed for highly sensitive tetracycline (TC) detection based on the peroxidase-like activity of Fe3O4@Cu nanoparticles and "sandwich" oligonucleotide hybridization. The Fe3O4@Cu nanoparticles with high peroxidase-like activity were successfully synthesized under mild conditions. Then, a "sandwich" oligonucleotide hybridization probe (a short amino-modified complementary sequence of a portion of the TC aptamer (cDNA1), TC aptamers, and a long complementary to 5' terminal TC aptamer sequence (cDNA2)) was created in 96-wells plates via DNA hybridization in the absence of TC from the detection system. The unique "sandwich" oligonucleotide hybridization probe adsorbed large numbers of Fe3O4@Cu nanozymes while further enhancing its peroxidase-like activity. Based on the 3,3',5,5'-tetramethylbenzidine (TMB)-hydrogen peroxide (H2O2) reporting system, the blue color of the solution decreased linearly with the increase of TC concentration, ranging from 10-3 to 103 µg/L with an ultralow limit of detection (LOD) of 0.91 ng/L (2 pM). The proposed method was successfully applied to detect TC in spiked milk samples, with recoveries of 81.8 to 112%, demonstrating the excellent potential for highly sensitive TC detection in milk.

Corrigendum: Bazi Bushen Capsule Alleviates Post-Menopausal Atherosclerosis via GPER1-Dependent Anti-Inflammatory and Anti-Apoptotic Effects.

[This corrects the article DOI: 10.3389/fphar.2021.658998.].

Complete mitogenome of Phthorimaea operculella (Lepidoptera: Gelechioidea: Gelechiidae).

In this research, the complete mitochondrial genome (mitogenome) of Phthorimaea operculella was sequenced and annotated. The mitogenome of P. operculella is 15,269 bp in length and contains 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, 2 ribosome RNA (12s and 16srRNA) genes and 1 control region. In addition, we used Endoclita signifier as the outgroup to analyze phylogenetic relationship, and the phylogenetic tree showed the sister relationship between P. operculella and Tuta absoluta.

Bazi Bushen Capsule Alleviates Post-Menopausal Atherosclerosis via GPER1-Dependent Anti-Inflammatory and Anti-Apoptotic Effects.

Bazi Bushen capsule (BZBS), as a Chinese medicine used to relieve fatigue, has been proven effective for the treatment of atherogenesis through antilipid effects. To investigate the potential mechanism of BZBS in the anti-atherosclerotic effect, Ovx/ApoE-/- mice were applied to investigate the anti-atherosclerotic efficiency and potential mechanism of BZBS. Therapeutic effect was evaluated based on the number of CD68+ and CD3+ cells, the level of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), and the ratio of cleaved caspase-3/caspase-3, as well as increasing ratio of Bcl2/Bax. Human umbilical vein endothelial cells (HUVECs) were chosen to evaluate the role of GPER1. Treatment with BZBS reduced lipid deposition by reducing the numbers of CD68+ and CD3+ cells, the level of ICAM-1 and VCAM-1, and the ratio of cleaved caspase-3/caspase-3, and increasing the ratio of Bcl2/Bax as compared with the control group. In si-GPER1-treated HUVECs, the anti-apoptotic effect of BZBS was decreased. This study revealed that BZBS exhibited a clear effect against atherogenesis via GPER1-dependent anti-inflammatory and anti-apoptotic mechanisms. We believe that this manuscript is informative and useful for researchers pursuing the related alleviation of post-menopausal AS via anti-inflammatory and anti-apoptotic mechanisms.

Complete mitochondrial genome of Neotoxoptera formosana (Takahashi, 1921) (Hemiptera: Aphididae), with the phylogenetic analysis.

In this study, we sequenced and annotated the complete mitochondrial genome (mitogenome) of Neotoxoptera formosana (Takahashi) (Hemiptera: Aphididae). The complete mitogenome of N. formosana is 15,642 bp in length, and includes 13 protein-coding genes (PCGs), 2 ribosome RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and one control region. The overall base composition was as follows: 45.2% of A, 5.8% of G, 10.5% of C, and 38.4% of T, with a total of A + T content of 83.6%. The phylogenetic tree showed that N. formosana and Myzus persicae were clustered into one branch. This result will enrich the mitogenome of family Aphididae.

Chinese medicine Bazi Bushen capsule improves lipid metabolism in ovariectomized female ApoE-/- mice.

BACKGROUND: Dyslipidemia is one of the mechanisms of atherosclerosis (AS). Depletion of estrogen plays a key role in the pathogenesis of postmenopausal AS in women, and the blood lipid levels of women are closely related to endogenous estrogen levels. Phytoestrogens (PEs) exert estrogenic effects, including protection against AS, without the adverse effects of estrogen administration. Bazi Bushen capsule (BZBS) is a traditional Chinese medicine herbal compound prescription that has been shown to contain 11 unique PEs. In the present study, we assessed the effects of BZBS against lipid metabolism disorders. METHODS: All ApoE-/- mice underwent ovary ligation and bilateral ovariectomy (Ovx) to induce surgical menopause (Ovx/ApoE-/- mice), whereas the C57BL/6J mice underwent sham surgery (needle threading). Ovx/ApoE-/- mice were given a high-fat diet without estrogen and C57BL/6J mice were given a normal diet for 12 weeks. Ovx/ApoE-/- mice treated with G1, a highly selective G-protein-coupled estrogen receptor1 (GPER1) agonist with proven activity against AS, were used as positive controls. Estrogen levels were measured and uterine atrophy index was calculated to determine the success of the model. Serum levels of triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were measured in each group. The orthogonal projections to latent structures discriminant analysis (OPLS-DA) model was used to separate the groups, MetaboAnalyst was then used to analyze the metabolic pathway, and the most representative metabolites were finally identified. RESULTS: Removal of bilateral ovaries resulted in successful surgical menopause models, where BZBS increased estrogen levels but did not increase the risk of uterine proliferation. BZBS attenuated dyslipidemia, including decreased TG, TC, and LDL-C levels, but increased HDL-C levels. The OPLS-DA model successfully distinguished the groups with good predictive ability and revealed their tendency to separate from each other. MetaboAnalyst suggested that both the G1 group and high-dose BZBS (HD-BZ) could improve the effect of lipid metabolism: the glycerophospholipid metabolism pathway was mainly improved by the G1 group, while the inositol phosphate metabolism pathway was mainly improved by the HD-BZ group. For the four compounds with the highest content, the concentrations of docosahexaenoic acid (DHA), 3-hydroxybutyric acid, and 5(Z), 8(Z), 11(Z)-eicosatrienoic acid were dramatically lower in the model group compared to the control group. Lysophosphatidylethanolamine (18:0) was higher in the model group than in the control group. BZBS corrected these effects. CONCLUSIONS: BZBS treatment reduced serum lipid levels and improved fatty acid metabolism in high-fat diet-fed, surgically induced menopausal ApoE-/- mice.

Increased Expression of lncRNA UCA1 and HULC Is Required for Pro-inflammatory Response During LPS Induced Sepsis in Endothelial Cells.

Systemic uncontrolled inflammatory response, also termed as sepsis, is responsible for many mortalities. Bacterial endotoxin, lipopolysaccharide (LPS), is a major cause of sepsis in endothelial cells. Even though a lot of research has been done to define underlying mechanisms of LPS induced sepsis, the role of long non-coding RNAs (lncRNAs), a group of >200 kb RNAs in sepsis is not well-defined. Expression of pro-inflammatory mediators IL6, ICAM1, and VCAM1 (which encodes interleukin-6, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1, respectively) were determined following LPS treatment of human dermal microvascular endothelial cells (HMECs) for 24 h to confirm sepsis induction. RNA immunoprecipitation (RIP) analysis was performed using the chromatin modifying proteins (CMPs), heterogeneous nuclear ribonucleoprotein (hnRNP) K and corepressors of the RE-1 silencing transcription factor (coREST) as individual baits. Quantitative real time polymerase chain reaction (qRT-PCR) was performed on RNA isolated from immunoprecipitated pellets for six different lncRNAs. The effect of the differentially expressed lncRNAs were determined by ectopic overexpression of the lncRNAs before induction of sepsis. Expression of IL6, ICAM1, and VCAM1 were significantly upregulated following treatment of the HMECs with LPS for 24 h confirming induction of sepsis. RIP and qRT-PCR analysis revealed that the lncRNAs HULC, UCA1, and MALAT-1 were significantly enriched with the CMPs after sepsis. RNA interference using siRNAs targeting HULC and UCA1, but not MALAT-1, decreased the expression of IL6, ICAM1, and VCAM1 to endogenous levels. Our results were further validated in an in vivo model of sub-lethal LPS-induced sepsis, whereby siRNA mediated knockdown of UCA1 and HULC lncRNAs prevented induction of VCAM1, ICAM1, and IL6, as assayed by immunohistochemistry. Cumulatively, these results suggest that LPS induced in vitro sepsis in endothelial cells and induction of pre-inflammatory mediators are at least in part due to increased expression of the UCA1 and HULC lncRNAs.

[An application of arterial pressure-based cardiac output measurements in fluid management strategies of critically ill patients].

OBJECTIVE: To discuss the clinical significance of fluid management of severe patients according to arterial pressure-based cardiac output (APCO) monitoring volume responsiveness index. METHODS: A retrospective cohort study was conducted. The severe patients were selected from the intensive care unit (ICU) of the First Hospital of Jilin University from June 1st, 2012 to December 31st, 2013. The hemodynamic parameters were monitored by APCO, and the fluid resuscitation was managed by stroke volume variation (SVV) and passive leg-raising test (PLR) when the acute physiology and chronic health evaluation II (APACHEII) score ≥ 15, heart rate >100 bpm with the result that the preload and heart function could not be evaluated. The heart rate, SVV, lactic acid (Lac) and central venous pressure (CVP) and curative effect were recorded before and after carrying out fluid management strategy. The criteria of clinical effective was defined as heart rate decreased and (or) stroke volume (SV) increased ≥ 10%, accompanied by blood Lac and SVV decreased, other than, the cases did not meet above criteria were considered ineffective. RESULTS: Sixty-eight patients were enrolled in the study. (1) Before carrying out fluid management strategy: 40 cases with CVP>12 cmH2O (1 cmH2O=0.098 kPa), and 16 cases with 5-12 cmH2O, 12 with <5 cmH2O. SVV>13% in 35 cases, SVV <13% in 9 cases. PLR positive in 18 cases, and PLR negative in 6 cases. It was implicated that the patients with poor preload (SVV>13% and PLR positive) accounted by 77.9% (53/68). (2) There were 49 effective cases and 19 ineffective cases 4 hours after carrying out fluid management strategy, and the effective rate was 72.06% (49/68). While there were 56 effective cases and 12 ineffective cases after 12 hours, and the total effective rate was 82.35% (56/68). (3) In effective group, heart rate, SVV, Lac after fluid management strategy were significantly lower than those before fluid management strategy [4 hours after fluid management strategy: heart rate (bpm) 112.45 ± 13.53 vs. 129.55 ± 15.49, SVV (15.47 ± 6.32)% vs. (21.20 ± 7.40)%, Lac (mmol/L) 4.16 ± 3.12 vs. 6.21 ± 4.11; 12 hours after fluid management strategy: heart rate (bpm) 110.02 ± 13.92 vs. 129.61 ± 14.93, SVV (14.61 ± 15.52)% vs. (20.66 ± 7.40)%, Lac (mmol/L) 3.35 ± 2.26 vs. 6.11 ± 4.02, P<0.05 or P<0.01], while there was no significant difference in those markers between before and after fluid management strategy in ineffective group [4 hours after fluid management strategy: heart rate (bpm) 119.53 ± 11.68 vs. 125.79 ± 11.58, SVV (16.95 ± 6.48)% vs. (18.47 ± 4.96)%, Lac (mmol/L) 5.55 ± 3.80 vs. 6.54 ± 3.72; 12 hours after fluid management strategy: heart rate (bpm) 115.92 ± 11.71 vs. 123.40 ± 11.59, SVV (17.17 ± 6.09)% vs. (19.42 ± 8.25)%, Lac (mmol/L) 6.33 ± 3.40 vs. 7.21 ± 3.81, all P>0.05]. CVP only at 12 hours after fluid management strategy in effective group was significantly higher than that before fluid management strategy (cmH2O: 12.8 8 ± 3.38 vs. 11.27 ± 4.97, P<0.05). CONCLUSIONS: SVV monitored by APCO is a good indicator of volume responsiveness index, which can be used as an important reference combined with PLR for fluid management of severe patients.

Enhancing effect of hydroxypropyl-ß-cyclodextrin on the intestinal absorption process of genipin.

The purpose of this work is to investigate the effect of the genipin/hydroxypropyl-ß-cyclodextrin (HP-ß-CD) inclusion complex on the intestinal absorption of genipin and identify its mechanism of action. The phase solubility profile was classified as A(L) type, indicating the formulation of a 1:1 stoichiometry inclusion complex. Fourier transform infrared spectroscopy, Differential scanning calorimetry, X-ray powder diffractometry, and (1)H nuclear magnetic resonance (NMR) and two-dimensional (2D) (1)H rotating-frame Overhauser enhancement (ROESY) NMR spectroscopies further confirmed the formulation of the inclusion complex with superior dissolution properties than the drug alone. The results of single-pass intestinal perfusion showed that the intestinal absorption of genipin was affected by P-glycoprotein (Pgp). The absorption rate and permeability value of the inclusion complex were significantly higher than the free drug, suggesting that its enhancing effect was involved in its solubilizing effect and Pgp inhibitory effect. The mechanisms of HP-ß-CD on Pgp inhibition were demonstrated by restraining the Pgp ATPase activity rather than changing the fluidity of the cell membrane.