Synergistic effects of methotrexate and piperine on human osteosarcoma cell lines – An in-vitro study

Background: Osteosarcoma is a malignant cancer that effect bone and metastasizing to many vital organs such as lungs. There are many available drugs to treat the disease including tamoxifen, methotrexate (MTX), and cisplatin which have their own side effects and hurdles to become drugs of choice for the disease. On the other hand, introduction of herbal drugs as chemotherapeutic agents opened up new arena to potentiate the existing treatment by exhibiting synergy. Piperine (PPN) is widely used drug as anti-cancer agent as well as it has anti-inflammatory, analgesic properties, and also used in the treatment of abdominal pains, tuberculosis, arthritis, and respiratory illness. Aims and Objective: Thus, this study was designed to investigate the synergistic inhibitory potential of PPN and MTX on the MG63 osteosarcoma cell lines in vitro. Materials and Methods: The cell lines were cultured on DMEM medium and investigated for cytotoxicity of the drugs using MTT assay at 540 nm in UV. Three groups of cell lines administered with PPN, MTX, and PPN+MTX (1:1) in various concentrations and IC50 values were calculated based on the % cell viability graphs. Results: Results showed that the IC50 of PPN was 38.65, MTX was 123.98, and PPN+MTX was 15.13 proving the significant synergistic cytotoxic effect of PPN and MTX in inhibiting the proliferation of MG63 cell lines. Conclusion: Further research needs to be conducted in this field to elucidate the synergistic pathways in which PPN has shown a better anti-osteosarcoma effect when combined with MTX.

In vitro release study on self-microemulsion of curcumin and piperine / 药学实践杂志

Objective To study the release profile of curcumin and piperine from the compound self-microemulsion. Methods The release of curcumin and piperine in vitro was investigated by dynamic dialysis under the condition of phosphate buffer of pH 4.8 and 7.5 with 0.75% Tween-80. Results The cumulative release rates of curcumin in pH 4.8 and pH 7.5 were 94.85% and 84.38% in 108 h, respectively. The cumulative release rates of piperine were 92.85% and 90.05% in 36 h, separately. Conclusion Curcumin and piperine in self-microemulsion have sustained release properties and released more in the acidic environment similar to the environment in tumors.

Research progress of pharmacological action and mechanism of piperine / 中国药房

Piperine is a kind of amide alkaloids presenting in Piper nigrum L.，which has the pharmacological action such as protecting cardiovascular system ，regulating glucose and lipid metabolism ，anti-tumor，improving nervous system diseases ， anti-inflammation and so on. This paper summarized the pharmacological action and mechanisms of piperine in recent years and found that piperine ，as the main active ingredient of P. nigrum ，could protect the cardiovascular system by reducing inflammation and oxidative stress ；improve mitochondrial function through anti-inflammatory and antioxidant effects ，thereby regulate glucose and lipid metabolism ；play an anti-tumor role by mediating the signaling pathways of Wnt/β-catenin，NF-κB/Nrf-2/KeAP-1/HO-1， PI3K/Akt，TGF-β1/Smad2/ERK1/2；improve neurological diseases by inhibiting autophagy ，relieving inflammation ，improving antioxidant，inhibiting neuronal apoptosis and regulating the expression of related proteins in neurons ；play an anti-inflammatory effect by inhibiting the activity of NF-κB and other signaling pathways and reducing the expression of inflammation-related proteins. However，the mechanism of action of piperine is not perfect ，and most of the studies have been confined to the pharmacological level or a certain signaling pathway and a certain target ，without being able to elucidate the interconnection between the relevant signaling pathway and the specific target from a holistic perspective. In the follow-up ，the specific targets of piperine can be identified and clinical trials can be carried out to provide support for the clinical application of piperine.

Effects of piperine on experimental colon carcinogenesis induced by DMH/DSS and its mechanism / 中国医师杂志

Objective:To investigate the effects of piperine on experimental colon carcinogenesis induced by 1, 2-dimethylhydrazine (DMH)/sodium dextran sulfate (DSS) and its mechanism.Methods:36 mice were divided into control group, model group and piperine group, 12 mice in each group. The control group was given normal saline by gavage; The model group and piperine group were given 3.6 mg/(kg·d) of normal saline and piperine respectively after establishing the experimental colon cancer model induced by DMH/DSS. Tumor load and volume were observed. Hematoxylin and eosin (HE) staining was used to observe the histological change of colon in mice. RAS/PI3K/AKT related pathway protein expression was detected by Western blot.Results:The body weight gain, protein expression levels of cleaved poly-ADP ribose polymerase (PARP), cleaved caspase-3 in model group were significantly lower than those in the control group (all P<0.05). The protein expression levels of Bcl-2, Bax, pan-Ras, p-MEK, p-ERK, PI3K, p-AKT, NF-κBP65, c-Myc and cyclin D1 in model group were significantly higher than those in the control group (all P<0.05). The body weight gain, protein expression levels of cleaved PARP and cleaved caspase-3 in piperine group were significantly higher than those in model group (all P<0.05). The protein expression levels of Bcl-2, Bax, pan-Ras, p-MEK, p-ERK, PI3K, p-AKT, NF-κBP65, c-Myc and cyclin D1 in piperine group were significantly lower than those in model group (all P<0.05). Conclusions:Piperine can inhibit the occurrence of experimental colon cancer induced by DMH/DSS, which may involve multiple components of RAS/PI3K/AKT signal axis.

Piperine suppresses growth and migration of human breast cancer cells through attenuation of Rac1 expression

Objective: To investigate the effect of piperine on human breast cancer cells. Methods: The effect of piperine on proliferation and migration of human breast cancer cells, MCF-7 and MDA-MB-231, was investigated using colony formation assays, wound healing assays, Matrigel migration assays, flow cytometry, RT-qPCR, and Western blotting assays. Results: Piperine inhibited the growth of MCF-7 and MDA-MB-231 cells and suppressed colony formation. Cell reduction at the G 0 / G 1 phase and cell arrest at the G 2 /M phase were observed in breast cancer cells. However, the significant effect was only demonstrated in MDA-MB-231 cells. Moreover, cancer cell migration was suppressed by piperine at low concentration. RT-qPCR and Western blotting assays showed that piperine downregulated Rac1 gene and protein expression. Conclusions: Piperine could inhibit growth and migration of breast cancer cells by reducing Rac1 gene and protein expression.

Bioavailability augmentation of silymarin using natural bioenhancers: An in vivo pharmacokinetic study

Abstract Pharmacokinetic studies were carried out in male and female rats to quantify silymarin as silybin (A+B) after the oral administration of various silymarin formulations combined with three bioenhancers, namely, lysergol, piperine, and fulvic acid, and compared with plain silymarin formulation (control). A non-compartmental analysis, model independent analysis, was utilized, and various pharmacokinetic parameters (C max, T max, and AUC 0-t) were calculated individually for each treatment group, and the values were expressed as mean ± SEM (n = 6). Plasma samples obtained from the rats were analyzed for the concentration of silymarin through a validated RP-HPLC method and on the basis of data generated from the pharmacokinetic studies. Results indicated that the bioenhancers augmented pharmacokinetic parameters and bioavailability increased 2.4-14.5-fold in all the formulations compared with the control. The current work envisages the development of an industrially viable product that can be further subjected to clinical trials and scientifically supports the development of silymarin as a contemporary therapeutic agent with enhanced bioavailability and medicinal values.

In vitro skin retention and drug permeation study of Tongluo-Qutong rubber plaster by UPLC/UV/MS/MS

Abstract Tongluo-Qutong rubber plaster (TQRP), a typical Chinese patent medicine that contains 13 different herbal remedies, is widely used in clinical practice for the treatment of cervical spondylosis and osteoarthritis. However, due to a lack of in vitro transdermal studies, the active ingredients of TQRP have not been fully elucidated. This presents a huge obstacle for quality evaluation, pharmacokinetic studies and clinical safety assessment of TQRP. In this work, a UPLC/UV/MS/MS method was established and validated to evaluate five analytes in TQRP. The validation demonstrated linearity (r > 0.99), specificity (no co-eluting peaks at the retention times of the analytes), and precision (RSD < 15%) within acceptable parameters. A skin permeation study was performed to determine the concentrations of drugs delivered to the dermis. The 24-hour cumulative permeation of ferulic acid, aleo-emodin, emodin and piperine were 303.68, 709.31, 671.06 and 25561.01 ng/cm2, respectively. According to the fitting data of the TQRP active components, skin permeation was mainly due to a combination of passive diffusion and drug release after matrix erosion

Comparative study on in vivo and in vitro permeability of Huoxue Zhitong gel patch and microemulsion gel / 中国中药杂志

This study aims to establish a method for determination of paeonol(Pae), eugenol(Eug), and piperine(Pip) content in receptor liquid and research on the permeability and pharmacokinetics of Huoxue Zhitong gel patch and microemulsion gel. The Franz diffusion experiment was conducted to assess the percutaneous permeability, and the microdialysis method was employed to assess pharmacokinetics of Huoxue Zhitong gel patch and microemulsion gel. The content of Pae, Eug, and Pip in receptor liquid in vitro and in vivo was determined by HPLC and UPLC-MS. The Q_n and J_(ss) of Pae, Eug, and Pip in the gel patch were significantly higher than those in the microemulsion gel, indicating that the drug release was faster in the gel patch. The C_(max), AUC_(0-760), and MRT of Pae, Eug, and Pip in the gel patch were higher than those in the microemulsion gel, indicating that the gel patch can promote the penetration and prolong the skin residence of the drug. The results of this study provide reference for improving the dosage form of Huoxue Zhitong patch.

Determination of piperine in Anar-5 tablets by high performance liquid chrotamography and method validation / Монголын Анагаах Ухаан

Introduction@#One of the main ingredients of Anar-5 tablets is Piper longium L. Piperine alkaloids are the main active ingredients of the Piper longum and have anti-inflammatory, antioxidant and gastric protection properties.In the framework of the standardization study of Anar-5 tablets, a method was developed to determine the content of piperine in highly perpormance liquid chromatography, and then it was sought to include it in the method of analysis of Anar-5 drugs.@*Goal@#Quantitative determination of piperine in Anar-5 tablets and validate the method@*Material and Methods@#The research was conducted in the Chemistry and Chemical Technology Laboratory of the Research Center of the Institute of Traditional Medicine and Technology. And Anar-5 tablets (serial number 04012020) that are produced for experimental were used in the research. The standard substance, piperine alkaloids, was purchased from Green Chemistry.Purification of HPLC (organic solvent methanol, 99.9%, distilled water) was used. The EX 1600 HP/ PUMP high-performance liquid chromatography instrument (column Arcus EP+-C18, 5µm, 4.6x250 mm) and the organic solvent filter 0.45 μm syringe filter were used. The methodology related to this research was discussed and approved at the online meeting of the Ethics Committee of the Academy of Sciences on January 26, 2021. SPSS 16 software was used to statistically program the survey results.@*Results @#According to the above method, the retention time of the standard piperine is 10.38± 0.02 minutes, and the retention time of the piperine in Anar-5 tablets is 10.42±0.033 minutes. Relative velocity deviation RSD 1.077%, accuracy 0.65446±0.0068mg, stability 0.61298±0.013mg, capture time 10.42±0.033 minutes, relative standard deviation RSD≤2%, specificity 10.35 minutes, The equation of a line constructed with a standard curve is y=43360x-33587 and the correlation coefficient R2=0.9989. The piperine content of Anar-5 tablets was determined to be 0.61298±0.013 mg. The LOD and LOQ for piperine were in 2.268 μg/ml and 6.873 μg/ml, respectively.@*Conclusion@#The content of piperine in Anar-5 tablets can be determined by the HPLC method, and the appropriate conditions for this method have been established. The HPLC method is unique, accurate, linear, and stable, and meets ICH Q2 (R1) guideline criteria.

Natural compounds as potential inhibitors of SARS-CoV-2 main protease: An in-silico study

Objective:To explore natural compounds as potential inhibitors against main protease (Mpro) of SARS-CoV-2. Methods:In the current study, systematic molecular docking analysis was conducted using AutoDock 4.2 to determine the binding affinities and interactions between natural compounds and Mpro. Selected natural compounds were further validated using a combination of molecular dynamic (MD) simulations and molecular mechanic Poisson-Boltzmann surface area (MM/PBSA) free energy calculations. Results:Out of twenty natural compounds, four natural metabolites namely, amentoflavone, guggulsterone, puerarin, and piperine were found to have strong interaction with Mpro of SARS-CoV-2 based on docking analysis. During MD simulations, all four natural compounds bound to Mpro at 50 ns and MM/G/P/BSA free energy calculations showed that all four shortlisted ligands had stable and favorable energies with strong binding to Mpro protein. Conclusions:Guggulsterone is a potential inhibitor of COVID-19 main protease Mpro. Further in vitro and pre-clinical studies are needed.

Piperine treating sciatica through regulating inflammation and MiR-520a/P65 pathway / 中国天然药物

Although the etiology of sciatica remains uncertain, there is increasing evidence that the disease process of sciatica is associated with the levels of inflammatory factors. Piperine, an alkaloid isolated from Piper nigrum, has previously been demonstrated to inhibit inflammation and analgesic effects. The purpose of this study is to verify the regulatory relationship between miR-520a and p65 and to explore how miR-520a/P65 affects the level of cytokines under the action of piperine, so as to play a therapeutic role in sciatica. Through ELISA experiment, we confirmed that four inflammatory factors (IL-1β, TNF-α, IL-10, TGF-β1) can be used as evaluation indexes of sciatica. The differentially expressed miRNA was screened as miR-520a, by microarray technology, and the downstream target of miR-520a was P65 by bioinformatics. Real-time fluorescence quantitative PCR confirmed that the expression of miR-520a was negatively correlated with pro-inflammatory cytokines, positively correlated with anti-inflammatory cytokines and negatively correlated with p65 expression at mRNA level. The expression of p65 was positively correlated with pro-inflammatory cytokines and negatively correlated with anti-inflammatory cytokines at the protein level verified by ELISA and Western blot. HE staining analysis showed that the nerve fibers were repaired by piprine, the vacuoles were significantly reduced, and the degree of nerve fiber damage was also improved. Immunohistochemical analysis showed that the expression of p65 decreased after administration of piperine. Dual-luciferase reporter gene assay confirmed that the luciferase signal decreased significantly after cotransfection of miR-520a mimics and p65 3'UTR recombinant plasmid. To sum up, in the rat model of non-compressed lumbar disc herniation, piperine plays a significant role in analgesia. MiR-520a can specifically and directly target P65, and piperine can promote the expression of miR-520a, then inhibit the expression of p65, down-regulate the pro-inflammatory factors IL-1β and TNF-α, and up-regulate the effects of anti-inflammatory factors IL-10 and TGF-β1, so as to treat sciatica.

Inclusion of biopiperine in the kappa-carrageenan complex might improve its bioaccessibility and in vivo anti-inflammatory activity in edema-induced Wistar rats

Piper nigrum has been used in Indonesian traditional medicine to alleviate pain. Piperine, a nitrogenous substanceisolated from the plant, has been reported for its anti-inflammatory activity. However, this compound is slightlysoluble in water, which impacts its bioaccessibility. A recent study reported that a co-ground mixture of piperine andβ-cyclodextrin revealed a significant increase of dissolved piperine at 15 minutes of dissolution test compared to thatof pure piperine. This work was aimed to study the bioaccessibility of the carrageenan-complexed piperine in Wistarrats and assayed its anti-inflammatory activity on the edema-induced paw of the rats. Both isolated (from P. nigrum)and synthetic (TCI, Tokyo Chemical Industry) piperines were used as the standards for the bioaccessibility assay,whereas acetosal was the standard drug for the anti-inflammatory activity study. The carrageenan-complexed piperinerevealed a better bioaccessibility (Cmax = 0.34 µg/ml; Tmax at 30 minutes) than that of the isolated piperine (Cmax = 0.12µg/ml, Tmax at 60 minutes), whereas the synthetic piperine showed the best absorption (Cmax = 0.48 µg/ml, Tmax at 30minutes). The anti-inflammatory activity of carrageenan-complexed piperine at a dose of 393 mg/kg body weight(BW) (contains 100 mg of piperine) equals to the acetosal dose of 45 mg/kg BW. Thus, the inclusion of biopiperinein the carrageenan complex might improve its bioaccessibility and in vivo anti-inflammatory activity in Wistar rats.

Development And Validation Of Rp-Hplc Method For Simultaneous Estimation Of Gallic Acid, Curcumin And Piperine In An Ayurvedic Formulation

Objective: The objective of the present work was to establish a simple, precise, accurate and robust method for simultaneous estimation of gallic acid, curcumin and piperine from the marketed ayurvedic formulation by liquid chromatography. Methods: The separation was carried out on Hemochrom C18 Column (250 mm × 4.6 mm ID, 5 µm pore size) with a mobile phase methanol: acetonitrile: water (pH 3.2adjusted by using orthophosphate acid) in the ratio 70:20:10v/v by isocratic elution modeat 25 °C and the flow rate was setat0.8 ml/min. The analysis was carried out atisoabsorptive wavelength of 295 nm. Results: The retention time of gallic acid, curcumin and piperine was found to be 3.3(±0.2), 4.7 (±0.2) and 5.6 (±0.2) min, respectively. The linearity range for gallic acid, curcumin and piperine was found to be 10-70 μg/ml, 20-80 µg/ml and 2-14 µg/ml, respectively with the coefficient of linear regression greater than 0.99 for all markers. Mean percent recoveries for gallic acid, curcumin, and piperine were found within the limit of acceptance (99-100%). The percent relative standard deviation (%RSD) for precision and robustness was found less than 2%, which indicates the method is precise and robust. The developed method applied for quantification of these markers from the marketed ayurvedic formulation of Dekofcyn tablet. Conclusion: The developed method was found to be simple, rapid, precise and reproducible for standardization of Dekofcyn tablet and can be useful for other formulations containing these three markers.

Optimization of prescription process of curcumin-piperine polymeric micelles by central composite design-response surface method / 中草药

Objective: To optimize the prescription process of curcumin-piperine polymeric compound micelles (Cur/Pip F127/P123-PM) by central composite design-response surface method. Methods: The content of curcumin and piperine was determined by UPLC. The Cur/Pip F127/P123-PM was prepared by thin film hydration method. Based on the single factor test, the dosage, the mass ratio of F127 and the volume of water were used as independent variables, and the drug loading and entrapment efficiency of curcumin, entrapment efficiency of piperine and the micelle size were dependent variables, and next central composite design-response surface method of three factors and five levels was carried out. The analysis results showed and verified the optimal prescription. Finally, the optimal lyophilization conditions of the micelle preparation were initially screened. Results: The optimal preparation process was as follow: the dosage of curcumin and piperine was 12.96 mg and 0.69 mg, respectively; The mass ratio of F127 was 0.46, and the volume of water was 8.85 mL. The compound curcumin micelles prepared by the optimum formulation had the loading capacity of 5.63%, solubility of 1.27 mg/mL and entrapment rate of curcumin was 86.86%. The entrapment rate of piperine was 77.54%; The micelle size was 66.79 nm and the Zeta potential was close to zero. The lyophilized products prepared by using 8% mannitol as a protective agent had a good redispersion. Conclusion: The model established by central composite design-response surface method can be used to optimize the prescription of compound curcumin micelles, and the method had a high accuracy and good predictability advantage.

Content Determination of 4 Components in Different Parts of Mongolian Medicine Sugmel- 3 Decoction / 中国药房

OBJECTIVE：To establish a method for the content determination of apigenin and piperine in the water extract as well as eucalyptol and cumin aldehyde in the volatile oil of Mongolian medicine Sugmel- 3 decoction. METHODS ：HPLC method was adopted for the content determination of apigenin and piperine. GC method was used for the content determination of eucalyptol and cumin aldehyde. The determination of HPLC method was performed on Agilent Eclipse XDB-C 18 column with mobile phase consisted of methanol- 0.1％ phosphoric acid aqueous solution at flow rate of 1.0 mL/min；the detection wavelength was set at 225 nm（apigenin）and 342 nm（piperine）；the column temperature was set at 30 ℃ with sample size of 10 μL. The determination of GC method was performed on Dimensions SH-Rtx- 1 capillary column with high-purity hydrogen as carrier gas ； the injector temperature was set at 270 ℃，with flow rate of carrier gas 1 mL/min by temperature programmed ；the sample size was 1 μL，and split ratio was 1 ∶ 10. RESULTS：The linear ranges of apigenin ，piperine，eucalyptol and cumin aldehyde were 12.5-200 μg/g/mL（r＝0.999 6），87.3-139.7 μg/mL（r＝0.999 9），136-2 187 μg/mL（r＝0.999 9），39-635 μg/mL（r＝0.999 9）， respectively. The quantitation limits were 0.02，0.06，0.06，0.12 μg/mL，respectively. The detection limits were 0.01，0.02，0.03， 0.04 μg/mL. RSDs of precision，stability and reproducibility tests were all less than 4％. The recovery rates of the samples were 89.26％ -97.26％（RSD＝2.69％ ，n＝6），94.20％ -104.01％（RSD＝3.64％ ，n＝6），98.51％ -110.11％（RSD＝3.87％ ，n＝6）， 95.76％-107.82％（RSD＝4.12％，n＝6），respectively. The contents of above components were 0.769-0.828，7.741-7.981，5.284 7- 5.846 6，1.038 6-1.101 2 mg/g（n＝3）. CONCLUSIONS：The established method is simple and feasible ，and can be used for quality control of different parts of Mongolian medicine Sugmel- 3 decoction.

Autophagic activity of piperine on small intestine in dementia model mice with Parkinson's disease / 中国中药杂志

This article is to investigate the effect of piperine on the small intestine of mice with Parkinson's disease with dementia(PDD). Ninety-six C57 BL/6 mice of SPF grade were randomly divided into 8 groups(male, 12 in each group): normal group, model group, autophagy inhibitor group(6-amino-3-methylpurine, 3 MA, 30 mg·kg~(-1)), autophagy activator group(rapamycin, 1 mg·kg~(-1)), low, medium, and high dose piperine groups(10, 20, 40 mg·kg~(-1)), and medopar group(112.5 mg·kg~(-1)). Except for the normal group, mice in each group were injected subcutaneously with reserpine(0.1 mg·kg~(-1)) once every 48 hours for 40 days. In addition, on the 20 th day of administration, except for the normal group, the mice in the other groups were subjected to bilateral common carotid artery occlusion to finally prepare PDD models. At the same time, each group was given the corresponding drug treatment once a day for 40 days. After the last administration, the behavioral changes of mice were observed by autonomic activity experiment and hot plate experiment. The expression levels of α-synuclein(α-syn) and tyrosine hydroxylase(TH) in the small intestine were detected by immunohistochemistry. The expression levels of beclin-1, microtubule-associated protein 1 light chain 3 B(LC3 B) and p62 in the small intestine were detected by immunofluorescence assay. Hematoxylin-eosin staining was used to observe the pathological morphology of small intestine tissues in each group. Enzyme-linked immunosorbent assay was adopted for detection of β-amyloid precursor protein(APP), p-tau, acetylcholine transferase(ChAT), interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) in small intestine. Real-time fluorescent quantitative polymerase chain reaction was used to detect the expression of α-syn, TH, beclin-1, microtubule-associated protein 1 light chain 3(LC3), and p62 mRNA and mmu-miR-99 a-5 p in the small intestine. The results of this study showed that, as compared with the model group, the number of activities, the expression levels of ChAT, TH, and p62 were significantly increased in the 3 MA group, the various piperine dose groups, and the medopar group(P<0.05), and their first foot licking time was shortened; APP, p-tau, IL-6, TNF-α, α-syn, beclin-1, LC3 B and mmu-miR-99 a-5 p expression levels were significantly reduced(P<0.05). However, as compared with the model group, the number of activities, ChAT, TH, and p62 expression levels in the rapamycin group were significantly reduced(P<0.05), and the APP, p-tau, IL-6, TNF-α, α-syn, beclin-1, LC3 B and mmu-miR-99 a-5 p expression levels were significantly increased(P<0.05). As compared with the 3 MA group, the number of activities, ChAT, TH, and p62 expression levels were significantly reduced in the low and medium dose piperine groups and rapamycin group(P<0.05); howe-ver, their first foot licking time was significantly prolonged, APP, p-tau, IL-6, TNF-α, α-syn, beclin-1, LC3 B and mmu-miR-99 a-5 p expression levels were increased significantly(P<0.05). As compared with the medopar group, the number of activities, ChAT, TH, and p62 expression levels were significantly reduced in low dose piperine group and rapamycin group(P<0.05), but their first foot licking time was significantly extended, and APP, p-tau, IL-6, TNF-α, α-syn, beclin-1, LC3 B and mmu-miR-99 a-5 p expression levels were significantly increased(P<0.05). In addition, as compared with the normal group, the small intestinal epithelial cells of the model group and the rapamycin group were shed off a lot, with severe damages of intestinal mucosa as well as edema and shedding of the small intestine villi. After administration of the therapeutic interventions, the small intestinal epithelial cells of the 3 MA group, each dose group of piperine, and the medopa group were slightly damaged and the villi were slightly shed off. In summary, piperine has a protective effect on the small intestine of PDD model mice, showing reduced expression of mmu-miR-99 a-5 p, pro-inflammatory factors and autophagy factors, and the mechanism of slowing PDD pathological symptoms may be related to the inhibition of autophagy.

A rapid HPTLC method to estimate piperine in Ayurvedic formulations

Background: Trikatu, Sitopaladi, Hingavastaka, Avipattikara, Sringyadi and Talisadya are very popular Ayurvedic (churna) medicines practiced in India; however, unfortunately, they possess several qualitycontrol issues.Objective: The aim of this study was to find out a simple, accurate and sensitive HPTLC method for thedetection and quantification of marker molecule, piperine (alkaloid) on these Ayurvedic formulations forstandardization.Materials and methods: Methanolic extraction (reflux) was performed from the above six churnas as wellas three single ingredients Piper longum (pipul), Piper nigrum (marich) and Piper chaba (chai). HPTLC wasdone using piperine as a standard. The mobile phase was a mixture of toluene-ethyl acetate (7:3, v/v) anddetection at 342l.Results: The Rf was detected at 0.39. Piperine was quantified in all samples. P. nigrum showed higherpiperine than P. longum and P. chaba. The maximum piperine was noted in Hingavastaka churna andfollowed by Sringyadi churna, Sitopaladi churna, Talisadya churna, Trikatu churna and Avipattikara churna.Conclusion: This method can be successfully employed for standardization and quantitative analysis ofpiperine in Ayurvedic formulations (churnas) and also be helpful to clinicians and pharmacists to drawsignificant role of piperine present in all these samples.© 2017 Transdisciplinary University, Bangalore and World Ayurveda Foundation. Publishing Services byElsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Medicinal Impact of Piper- Nigrum (Piperine) Against Arsenic Induced Hepatic and Renal Toxicity in Experimental Mice

In view of the increasing risk of arsenic on human health, the present study has been carried out to investigate the hepato-protective effect of piperine on arsenic induced-hepatic and renal toxicity in mice. Various oxidative stress parameter, antioxidant level and micro nutrients were analyses in hepatic and hepatic renal organ of mice. Methods: Mice exposed arsenic (sodium arsenate 5 mg/kg body weight p.o. for 45 days) caused a significant increases oxidative stress in hepatic and renal tissue as compared to controls group. Results: Abnormal levels of arsenic in hepatic and renal tissue increased the levels of ROS, LPO, and decreased the levels of GSH with SOD, CAT, and GPx activities in the hepatic and renal tissue of mice as compared to controls. Co-treatment of arsenic with piperine (1.5 mg/kg body weight p.o. for 45 days) decreased the levels of ROS, LPO, and increased the level of GSH, also increased SOD, CAT, and GPx activity and showed improvements in hepatic and renal tissue of mice as compared to arsenic-treated groups. Conclusion: Our results proved that piperine worked as antioxidant, anti- inflammatory in nature.

Determination Of Piperine Content In Pepper By Using Different Organic Solvents: Through Hplc

Chromatography can be described as a mass transfer process involving adsorption using a nonpolar stationary phase and a mobile polar phase titrating through the column. The active component of the column, the sorbent or the stationary phase , is typically a granular material made of solid particles (e.g. silica, polymers, etc.,). The component of the sample mixture are separated from each other by means of mobile phase and different degrees of interaction with the sorbent particles based on their relative polarity. In the present study we have extracted piperine from grounded pepper using different chemicals such as petroleum ether, acetone and methanol. Petroleum ether extraction showed higher piperine content of 9.12% than methanol and acetone 3.15% and 3.37% respectively.

Anti-oxidative Effect of Piperine from Piper nigrum L. in Caenorhabditis elegans

Piper nigrum L. (Piperaceae), which is a well-known food seasoning, has been used as a traditional medicine for the treatment of vomiting, abdominal pain, diarrhea and anorexia in Korea, China and Japan. Methanol extract from the fruit of P. nigrum was successively partitioned as n-hexane, methylene chloride, ethyl acetate, n-butanol and H₂O soluble fractions. Among those fractions the ethyl acetate soluble fraction showed the most potent DPPH radical scavenging activity, and piperine was isolated from the ethyl acetate fraction. To know the antioxidant activity of piperine, we tested the activities of superoxide dismutase (SOD) and catalase together with oxidative stress tolerance and intracellular ROS level in Caenorhabditis elegans. To investigate whether piperine-mediated increased stress tolerance was due to regulation of stress-response gene, we quantified SOD-3 expression using transgenic strain including CF1553. Consequently, piperine enhanced SOD and catalase activities of C. elegans, and reduced intracellular ROS accumulation in a dose–dependent manner. Moreover, piperine-treated CF1553 worms exhibited significantly higher SOD-3::GFP intensity.