1.Effect and Mechanism of Berberine Hydrochloride Against Lung Cancer Cells in Vitro Based on Mevalonate Pathway
Yukun ZHENG ; Guangzhong WANG ; Xiaowei WU ; Qipan JIAN ; Yuhang SHU ; Zhongshi ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(13):92-101
ObjectiveTo investigate the efficacy and mechanism of berberine hydrochloride (BBH) against lung cancer cells through the mevalonate (MVA) pathway. MethodHuman lung cancer A549 cells and mouse Lewis lung carcinoma (LLC) cells were used as research subjects. Cell proliferation and cell counting kit-8 (CCK-8) assay were performed to detect the inhibitory effect of BBH (10, 20, 30, 40, 50 μmol·L-1) on the proliferation of the two kinds of cells (48 h). Then cell scratch assay was used to explore the influence of BBH (40 μmol·L-1) on the migration of A549 and LLC cells (24, 48 h), and colony formation assay was conducted to compare the colony formation ability of the cells under different concentrations of BBH (10, 20, 40 μmol·L-1). Moreover, the effects of BBH (40 μmol·L-1) on the content of acetyl-coenzyme A (A-CoA) and total cholesterol (TC) in A549 and LLC cells were determined by kit assay. AutoDock Vina was used for the dock of BBH and MVA pathway regulatory protein, sterol regulatory element-binding protein 2 (SREBP2). Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to observe the effects of BBH (40 μmol·L-1) on the mRNA expression of nine genes related to the MVA pathway in A549 and LLC cells: hydroxymethylglutaryl-CoA synthase 1 (HMGCS1), hydroxymethylglutaryl-CoA Reductase (HMGCR), mevalonate kinase (MVK), phosphomevalonate kinase (PMVK), mevalonate 5-pyrophosphate decarboxylase (MVD), farnesyl diphosphate synthase (FDPS), squalene epoxidase (SQLE), farnesyl-diphosphate farnesyltransferase 1 (FDFT1), and geranylgeranyl diphosphate synthase 1 (GGPS1). Western blot was performed to clarify the effects of BBH (40 μmol·L-1) on the expression of three key proteins of the MVA pathway: HMGCS1, HMGCR, and FDFT1. The Cancer Genome Atlas (TCGA) database was searched to analyze the relationship between HMGCS1, HMGCR, FDFT1 and transcription gene SREBF2 in non-small cell lung cancer (NSCLC). ResultCompared with the conditions in the control group, the proliferation, migration, and colony formation of A549 and LLC cells in the BBH group were decreased (P<0.01), while the cell apoptosis rate was increased (P<0.01). Molecular docking showed that BBH had good binding activity with SREBP2. In addition, the content of A-CoA and TC of the MVA pathway was reduced (P<0.01). BBH down-regulated the mRNA expression of HMGCS1, HMGCR, MVK, PMVK, MVD, FDPS, SQLE, FDFT1, and GGPS1 in A549 and LLC cells (P<0.01), and lowered the levels of HMGCS1, HMGCR, and FDFT1 proteins (P<0.05, P<0.01). In NSCLC patients, HMGCS1, HMGCR, and FDFT1 were highly correlated with SREBF2 (R=0.54, R=0.57, and R=0.48). ConclusionBBH can inhibit the proliferation, migration, and colony formation of A549 and LLC cells and promote cell apoptosis, which may be related to the regulation of MVA pathway by BBH binding to SREBP2.
2.Comparison of the fingerprints and in vitro antioxidant activity of decoction pieces of Polygonum cuspidatum by integrated processing and traditional processing
Qipan JIAN ; Yingrui ZHU ; Yukun ZHENG ; Huanbo CHENG ; Jiyuan TU ; Guangzhong WANG
China Pharmacy 2022;33(5):569-574
OBJECTIVE To compare t he diff erences of the fingerprint and in vitro antioxidant activity between decoction pieces of Polygonum cuspidatum by integrated technology of habitat processing and processing (short for IPDP )and traditional processing decoction pieces (short for TPDP ). METHODS Ten batches of IPDP and ten batches of TPDP were prepared by integrated technology and traditional technology ,respectively. HPLC method was used to establish and compare the fingerprints of IPDP and TPDP. The scavenging rates of DPPH free radical ,ABTS free radical ,superoxide free radical and hydroxyl free radical and reducing activity of Fe 3+ were detected for IPDP and TPDP. In vitro antioxidant activities were compared between IPDP and TPDP. RESULTS There were 11 common peaks in the fingerprints of IPDP and TPDP ,among which 17 came from IPDP and 13 came from TPDP. The peak heights of peak 6(polydatin)and peak 15(emodin-8-O-β-D-glucoside)in IPDP were significantly higher than those in the TPDP ,and the peak heights of peak 13(resveratrol),peak 17(emodin)and peak 19(physcion)in the TPDP were significantly higher than those in the IPDP. The results of in vitro antioxidant test showed that IPDP and TPDP had a certain scavenging capacity on DPPH free radical ,ABTS free radical ,superoxide free radical and hydroxyl free radicals ,and also had a certain reducing capacity on Fe 3+. CONCLUSIONS The integrated processing technology of P. cuspidatum has a good retention effect on the glycosides in P. cuspidatum ,and the in vitro antioxidant activity of IPDP is stronger than that of TPDP.
3.Comparison Between Integrated and Traditional Decoction Pieces of Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine Based on Quality and Pharmacological Effect
Caixia TAO ; Yukun ZHENG ; Qipan JIAN ; Yuhang SHU ; Jiyuan TU ; Yanju LIU ; Guangzhong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(23):174-181
ObjectiveTo compare the consistency and difference between the integrated decoction pieces (IDP) and traditional decoction pieces (TDP) of Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine (PRPZA) in the component content and pharmacological effect, so as to explain the rationality of the integrated production of PRPZA. MethodThin layer chromatography (TLC), extract determination, ultraviolet spectrophotometry and high performance liquid chromatography (HPLC) were used to analyze and determine the TLC identification, the contents of water-soluble extract, alum residue, needle-like calcium oxalate crystal, protein, total alkaloids, polysaccharides and three nucleosides (inosine, guanosine and adenosine) of IDP and TDP of PRPZA, and the statistical comparison was made. The anti-inflammatory effect and irritation of IDP and TDP of PRPZA were compared by xylene-induced mouse auricle swelling test and rabbit conjunctival irritation test. ResultCompared with the TDP of PRPZA, the contents of alum residue, needle-like calcium oxalate crystal, protein, polysaccharides, inosine, guanosine and the total amount of three nucleosides in the IDP of PRPZA decreased by 16.95%, 21.27%, 23.78%, 4.74%, 52.12%, 0.24% and 26.04%, the contents of water-soluble extract, total alkaloids and adenosine increased by 7.62%, 114.83% and 125.42%, respectively. IDP and TDP of PRPZA had obvious inhibitory effects on ear edema in mice, but there was no significant difference between them, indicating that the anti-inflammatory effect was consistent. The two decoction pieces of PRPZA had no irritation to the conjunctiva of rabbits, and the difference was not statistically significant between them, suggesting that the safety was similar. ConclusionThere is a certain difference in the component content between the IDP and the TDP of PRPZA, but their anti-inflammatory and irritant effects are similar, and the quality of IDP is slightly better than TDP, which provides a reference for the industrial production and clinical application of the IDP of PRPZA.
4.Historical Evolution and Modern Research Overview of Traditional Chinese Medicine Simmering Method
Zhenhui LI ; Benye WANG ; Yuhang SHU ; Qipan JIAN ; Jiyuan TU ; Yanju LIU ; Guangzhong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):293-298
Simmering method is one of the traditional processing methods of Chinese materia medica, which has been documented in the herbal literature and medical books of the past dynasties and has a great variety, but at present, there are not many specific varieties of Chinese materia medica involved, and there are few related researches. By reviewing the ancient and modern related information, the authors have organized and analyzed the historical evolution, processing purpose, modern representative Chinese materia medica(processing technology, quality evaluation, pharmacological research) of simmering method. After sorting out, it was found that the simmering method was widely used in ancient times, which was first seen in Huashi Zhongzangjing of the Eastern Han dynasty, and was enriched and developed through the Tang, Song, Jin, Yuan, Ming and Qing dynasties, and entered its heyday in Ming and Qing dynasties along with the economic prosperity and development of the Ming dynasty, involving as many as 159 varieties of Chinese materia medica, and gradually perfecting the processing theory of the simmering method. However, the number of varieties that still use the simmering method in modern times significantly decreased. The main purposes of using simmering method in modern Chinese materia medica are to reduce adverse reactions, moderate medicinal properties, enhance therapeutic effects, remove non-medicinal parts, and facilitate further processing, etc. This paper combed the key information of simmering methods for Chinese materia medica from ancient to modern times, which can provide a literature basis for the clinical application and modern research of simmered products of Chinese materia medica.