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1.
Chinese Pharmacological Bulletin ; (12): 1718-1724, 2023.
Article in Chinese | WPRIM | ID: wpr-1013705

ABSTRACT

Aim To investigate the role of autophagy in the dysfunction of testicular TM4 cell junction induced by ERα down-regulation. Methods TM4 cells were treated with different concentrations of E R a inhibitor ICI182780 (ICI), and the proliferative activity of TM4 cells was detected by CCK-8 method. The number and morphological changes of TM4 cells were observed by light microscope. The levels of E R a, junction function related proteins and autophagy marker proteins were detected by Western blot. The expression and localization of Cx43 were detected by immunofluorescence staining. The cells were treated with chloroquine (CQ) and ICI for 24 h. The expression levels of autophagy and junction function related proteins were detected by Western blot. Results When ICI concentration was 50 nmol • L ~ or above, the cell viability decreased significantly. The increase of cell vacuoles in ICI group was observed by light microscope. Compared with normal control group, the protein expression levels of E R a, ZO-1, occludin, claudin-11, p-catenin and Cx43 in ICI groups significantly dropped, while the expression levels of N-cadherin and E-cadherin had no significant changes; LC3 II significantly rose, while p62 expression significantly fell. The results of immunofluorescence showed that the fluorescence expression of Cx43 in ICI group decreased significantly, but the position of CX43 did not change significantly. Compared with ICI group, the expression levels of LC3 II, p62, Cx43, ZO-1 and β-Catenin significantly increased. Conclusions The down-regulation of E R a leads to damage of TM4 cell junction function, which may be related to the activation of autophagy.

2.
Chinese Pharmacological Bulletin ; (12): 2346-2353, 2023.
Article in Chinese | WPRIM | ID: wpr-1013677

ABSTRACT

Aim To study the effects of high-fat diet on testicular germ cell apoptosis in mice through endoplasmic reticulum stress. Methods C57BL/6J male mice were assigned into normal group and high-fat diet group randomly, with six mice in each group. The mice in normal group or high-fat diet group were fed with regular or high-fat diet continuously for five months. The mice were weighed, anesthetized, and euthanized to collect testicular and epididymal tissue for analysis. The testicular tissue was weighed and their indices were calculated. Epididymal tissue was collected for semen analysis. The morphological alterations of testicular tissue were observed using hematoxylin-eosin ( HE ) staining. The apoptosis of germ cells was detected by TUNEL staining and the apoptotic indices were calculated. The expression levels of apoptosis and endoplasmic reticulum stress-related proteins in testicular tissue were detected by Western blot. The protein expression and localization of GRP78 in testicular tissue were further detected by immunofluorescence. Results The results showed that compared to the normal group, the high-fat diet group had a significant increase in body weight, a significant decrease in testicular index, sperm concentration, and sperm vability, loose arrangement of germ cells, significant thinning of the seminiferous epithelium, no significant change in the diameter of seminiferous tubules, a significant increase in germ cell apoptosis , with an increased apoptosis index, and significant increase in expression of Bax and cleaved-caspase-12,and a significant decrease in Bcl-2 protein expression. The expression levels of GRP78 , p-IREl, XBP1, and ATF6a proteins were significantly up-regulated, while p-PERK, p-eIF2a, ATF4 protein expression showed no significant changes. Immunofluorescence results further showed a significant increase in the expression of GRP78 protein in the testicular tissue,with no significant changes in the expression location. Conclusions High-fat diet can induce the apoptosis of mouse testicular germ cells, and the mechanism may be related to the activation of endoplasmic reticulum stress IRE1 and ATF6 signaling pathway.

3.
Acta Pharmaceutica Sinica ; (12): 3408-3420, 2023.
Article in Chinese | WPRIM | ID: wpr-999085

ABSTRACT

In this study, the mechanism of Xiaoyan Lidan formula (XYLDF) against 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-collidine (DDC)-induced chronic intrahepatic cholestasis (CIHC) in mice was investigated based on metabolomics, molecular docking and pharmacological methods. In the pharmacodynamics study, a dosage of 5 g·kg-1 (clinical equivalent) XYLDF was administered in DDC-induced mice, then the effect of XYLDF against CIHC was evaluated by measuring the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP) as well as total bilirubin (TBIL) in serum and observing liver histopathological changes. All experiments were approved by the Ethical Committee Experimental Animal Center of Guangzhou University of Chinese Medicine (ZYD-2021-001). The serum metabolites of mice in each group were detected and identified based on ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry, and the relevant biological pathways and molecular key targets were further enriched. Molecular docking technology was used to further evaluate the binding activity of the main active ingredients of XYLDF with potential targets. Subsequently, the in vitro experiment was conducted for the validation of the vital target. The results showed that compared with the model group, XYLDF significantly decreased the levels of ALT, AST, AKP and TBIL in the serum of CIHC mice, as well as alleviated inflammatory infiltration and hepatocyte necrosis in liver tissue. According to the metabonomic study, a total of 35 differential metabolites was identified as biomarkers associated with cholestasis, 12 of which were significantly recovered by XYLDF treatment. These biomarkers were involved in the pathways of primary bile acid biosynthesis and linoleic metabolism, which are closely related to the mechanism of XYLDF against CIHC. Protein-protein interaction network indicated that cytochrome P450 3A4 (CYP3A4) and cytochrome P450 1A1 (CYP1A1) are significant potential targets with good binding properties with six major active ingredients of XYLDF. Furthermore, it was found that 4-methoxy-5-hydroxycanthin-6-one, dehydroandrographolide and isodocarpin, three of the main active components in XYLDF, markedly induced the expression of CYP3A4 mRNA in vitro. This study revealed that XYLDF mainly mediates the biosynthesis of bile acids in CIHC mice to improve liver tissue lesions and bile efflux disorders, among which, CYP3A4 is the key target in the protection of XYLDF against CIHC. This research provides a reference for further elucidation of the pharmacological mechanism of XYLDF.

4.
China Journal of Chinese Materia Medica ; (24): 1054-1065, 2023.
Article in Chinese | WPRIM | ID: wpr-970577

ABSTRACT

This study aims to examine the effect of superfine powder and aqueous extract of Polygonati Rhizomaon on natural perimenopausal syndrome in rats and explore the underlying mechanism. To be specific, a total of 60 female SD rats(14-15 months old) with estrous cycle disorder were screened by the vaginal smear and randomized into model control group, β-estradiol 3-benzoate group(0.1 mg·kg~(-1)), superfine powder of Polygonati Rhizoma group(0.25, 0.5 g·kg~(-1)) and aqueous extract of Polygonati Rhizoma group(0.25, 0.5 g·kg~(-1)), and another 10 female SD rats(14-15 months old) were selected as the youth control group. The administration lasted 6 weeks. Then the perimenopausal syndrome-related indexes such as body temperature, microcirculatory blood flow of face and ear, vertigo period, salivary secretion, grip force, and bone strength were determined and open field test was conducted. The immune system-related indexes such as the wet weight and index of thymus and spleen, percentage of T lymphocytes and subgroups in peripheral blood, and hematological indexes were measured. In addition, the ovary-related indexes such as estrous cycle, the wet weight and index of uterus and ovary, ovarian tissue morphology, and cell apoptosis were determined. Moreover, hypothalamus-pituitary-ovary axis(HPO)-related indexes such as serum sex hormone levels, cytochrome P450 family 11 subfamily A member 1(CYP11A1), cytochrome P450 family 19 subfamily A member 1(CYP19A1), and cytochrome P450 family 17 subfamily A member 1(P450 17A1) in ovarian tissue were measured. The results showed that the superfine powder and aqueous extract of Polygonati Rhizoma significantly decreased body temperature(anal, facial and dorsal temperature), microcirculatory blood flow in the ear, and vertigo period, increased salivary secretion, grip force, bone strength, total distance and total speed in the open field test, wet weight and index of thymus and spleen, lymphocyte ratio, CD3~+ level, and CD4~+/CD8~+ ratio, reduced neutrophil number and ratio, estrous cycle disorder ratio, and number of ovarian apoptotic cells, raised wet weight and index of uterus, wet weight of ovary, levels of inhibin B(INHB), estradiol(E_2), anti-müllerian hormone(AMH), and ovarian CYP11A1 and CYP19A1, decreased follicle-stimulating hormone(FSH) and luteinizing hormone(LH) content, and improved ovarian tissue morphology. It is suggested that the superfine powder and aqueous extract of Polygonati Rhizoma can improve the symptoms associated with natural perimenopausal syndrome in rats and enhance ovarian function and immune function. The mechanism is that they regulate HPO axis function by increasing estrogen synthesis.


Subject(s)
Female , Animals , Rats , Rats, Sprague-Dawley , Microcirculation , Cholesterol Side-Chain Cleavage Enzyme , Perimenopause , Powders , Cytochrome P-450 CYP1A1
5.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 183-192, 2021.
Article in Chinese | WPRIM | ID: wpr-906346

ABSTRACT

Objective:To explore the potential molecular mechanism of corylin in the treatment of lung cancer. Method:A549 cells were treated with different concentrations of corylin, and their proliferation was detected using methye thiazolye telrazlium (MTT) reagent. Then the trend analysis and gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were conducted to screen the key genes and pathways of corylin against A549 cell proliferation, followed by the verification of sequencing results by real-time polymerase chain reaction (Real-time PCR). Result:Corylin inhibited the proliferation of A549 cells and regulated the expression of 4 364 genes in cells. The trend analysis revealed that these genes were clustered into 20 distinct modules, among which four were significantly down-regulated, suggesting that corylin exerted the anti-proliferation effect by inhibiting the expression of some genes. The inter-group comparison of differentially expressed genes (DEGs) showed that the elevation in the concentration of corylin resulted in more down-regulated genes but weakened proliferation, consistent with the findings by trend analysis. The GO and KEGG enrichment analysis of 278 DEGs in the high-dose corylin group demonstrated that corylin mainly changed the cellular and metabolic processes, which was attributed to its regulation of steroid biosynthesis, fatty acid metabolism, biosynthesis of unsaturated fatty acids, synthesis and degradation of ketone bodies, and steroid hormone biosynthesis. The Real-time PCR results confirmed that corylin down-regulated the mRNA expression levels of LSS, stearoyl-CoA desaturase (SCD), 3-hydroxy-3-methylglutaryl-CoA synthase 1 (HMGCS1), but up-regulated the mRNA expression of recombinant human angiopoietin-like protein 4 (ANGPTL4), basically consistent with the transcriptomics results. Conclusion:Corylin inhibits A549 cell proliferation and alleviates lung cancer by targeting the related genes in lipid metabolism pathways.

6.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 240-250, 2021.
Article in Chinese | WPRIM | ID: wpr-905855

ABSTRACT

Periplaneta americana is one of the important basic medicinal materials of traditional Chinese medicine "fei lian". The traditional functions mainly include promoting blood circulation, sore muscles, diuresis, spleen and phlegm. Because of its exact curative effect, proprietary Chinese medicines, which are mainly used as raw materials, are widely used in clinical practice, especially in the repair of various wounds. The drug has not been included in the Chinese Pharmacopoeia. The local standard is only based on the alanine content of total amino acids. The physiologically active small peptides, nucleosides, proteins and other substances have not been obtained. Qualitative or quantitative control. In recent years, peptide monomers isolated from the P.americana, such as antimicrobial peptides, neuropeptides, and diuretic peptides, have strong pharmacological activities such as antibacterial, antitumor, and muscular neurotrophic, and dihydroisocoumarins are also irritating. Dermal Dermal fibroblasts produce collagen. Based on this, this paper uses CNKI, Wanfang Database and Pubmed Database to search the relevant research literatures of P.americana from 1984 to 2019, and systematically analyzes the current research of P.americana from three aspects: chemical composition-pharmacological action-clinical application. Interpretation provides reference for the further development of the drug and the development of more specific and stable quality control standards for its proprietary Chinese medicines.

7.
China Journal of Chinese Materia Medica ; (24): 1667-1673, 2021.
Article in Chinese | WPRIM | ID: wpr-879076

ABSTRACT

This study aims to investigate the preventive effect of Dendrobium officinale in LPS-induced intestinal mucosal damage. Forty SPF-grade C57 BL/6 J male mice were randomly divided into normal group(NC), model group(LPS), and two superfine powder groups of Dendrobium officinale(DOF)(DOF-L, 0.30 g·kg~(-1)and DOF-H, 0.60 g·kg~(-1), respectively), with 10 mice in each group. DOF superfine powder suspension was given via oral administration to mice for 7 days, while the mice in NC and LPS groups received the same volume of saline for 7 days. On the eighth day, the mice in LPS group and DOF treatment groups were injected with LPS(5 mg·kg~(-1)) by intraperitoneal injection to establish the intestinal mucosal injury model, while the mice in NC group were injected with the same volume of sterile saline in the same manner. Six hours after injection with LPS or saline, plasma and the intestinal tissue were collected. The diamine oxidase(DAO) and D-lactate levels in plasma were detected with a biochemical method. The levels of proinflammatory factors interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) in plasma were detected by ELISA. The histomorphology and ultrastructure of mouse ileum tissues were observed by hematoxylin-eosin(HE) staining in optical microscope and transmission electron microscope(TEM). The expression and distribution of tight junction(TJ) proteins claudin-1, occludin and F4/80 were detected by immunohistochemistry while the protein expression levels of Toll-like receptor 4(TLR-4) and nuclear factor kappa B p65(NF-κB p65) in jejunum were detected by Western blot. The experimental results showed that continuous intragastric administration of D. officinale superfine powder for 7 days obviously alleviated the damage and ultrastructural changes of intestinal mucosa induced by LPS; significantly decreased DAO and D-lactate levels in plasma in model group(P<0.05); up-regulated the protein expression of claudin-1 and occludin in ileum tissues; down-regulated the protein expression of TLR-4 and NF-κB p65 in jejunum tissues(P<0.01); significantly decreased TNF-α and IL-6 levels in plasma(P<0.05); and decreased the infiltration of F4/80~+ macrophage cells. Our results suggested that D. officinale had significant protective effects on LPS-induced intestinal mucosal damage and reduced intestinal permeability. The mechanism might be related to its effects of inhibiting inflammation via TLR-4/NF-κB p65, and up-regulating the expression of tight junction proteins.


Subject(s)
Animals , Male , Mice , Dendrobium , Intestinal Mucosa , Lipopolysaccharides , NF-kappa B , Powders , Tumor Necrosis Factor-alpha/genetics
8.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 86-91, 2020.
Article in Chinese | WPRIM | ID: wpr-862664

ABSTRACT

Objective::To observe effect of addition and subtraction therapy of Huaihuasan combined with Taohuatang to ulcerative colitis with cold-heat complicated syndrome at active stage, and to study regulation effect to immune function and inflammatory response. Method::One hundred and twelve patients were randomly divided into control group and observation group by random number table. Patients with light and middle symptoms in control group got mesalazine slow release tablets, 1.0 g/time, 3 times/days, patients with severe symptoms or whose symptoms were not changed after getting for 4 weeks in control group got prednisone acetate tablets, 0.75 mg·kg-1·d-1 for 3 times. Based on the treatment in control group, patients in observation group added Huaihuasan combined with Taohuatang, 1 dose/day. The course of treatment was 4 weeks. At remission period, mesalazine slow release tablets were used for maintain long-term maintenance therapy, 0.5 g/times, 3 times/days. Scores of disease activities were graded by improvement mayo, and clinical remission and clinical efficacy were recorded, scores of cold-heat complicated syndrome, mucous membrane under enteroscopy and histology of mucosa belongs to Geboes were graded. And levels of tumor necrosis factor-α(TNF-α) in peripheral blood, interleukin-8 (IL-8), IL-10, T lymphocyte subsets (CD4+, CD8+), and adverse reactions, 6 months' follow-up and recurrence were also be recorded. Result::Therapeutic effect of traditional Chinese medicine syndromes were analyzed by rank sum test, which in observation group was better than that in control group (Z=1.915, P<0.05). Clinical effect in observation group was 98.04%(50/51) higher than 84.00%(42/50) in control group, clinical remission rate was 94.12%(48/51) higher than 78.00%(39/50) in control group, and mucosal healing rate was 96.08%(49/51) higher than 82.00%(41/50) in control group (P<0.05). Scores of mayo, cold-heat complicated syndrome, colonic mucosa and index scores of Geboes were all lower than those in control group (P<0.01). And levels of TNF-α, IL-8 and CD8+ were lower than those in control group (P<0.01), and levels of IL-10, CD4+ and CD4+ /CD8+ were higher than those in control group (P<0.01). Recurrence rate during 6 months in observation group was 10.42%(5/48) lower than 30.77%(12/39) in control group (χ2=5.669, P<0.05), as for adverse reactions, there was no significant difference between two groups. Conclusion::Huaihuasan combined with Taohuatang can induce UC to remission period, inhibit the activity of disease, alleviate clinical symptoms, regulate immune function and expression of inflammatory factors, alleviate inflammatory reaction, promote intestinal mucosal healing, and can maintain clinical remission and reduce recurrence. The clinical efficacy is superior to that of 5-ASA/glucocorticoid in Western medicine.

9.
Chinese Medical Equipment Journal ; (6): 72-74, 2018.
Article in Chinese | WPRIM | ID: wpr-699970

ABSTRACT

Objective To investigate the application of SIEMENS Ysio DR amplification in the radiation dose reduction. Methods Totally 160 patients with similar heights and body shapes were divided equally into A,B,C and D groups,and then underwent radiological examinations with SIEMENS Ysio DR from September 2014 to May 2016.A and B groups went through right hand AP oblique examination with the same kV while different mAs, and C and D groups had chest posteroanterior examination with different kV and the same mAs. Image quality was regulated with the amplification coefficient, and then evaluated with double-blind method by two senior radiologists.Statistical analysis was executed over the values of mAs in A and B groups and those of kV in C and D groups. Results A group had the mAs value being(2.26 ±0.16)mAs, which was significantly different from that of B group [(4.05±0.19)mAs] (P<0.05);while C group had kV value being (104.79±6.39)kV, which was statistically different from that of D group [(83.48±3.67)kV] (P<0.05). Satisfactory image were obtained by regulation with amplication coefficient.There was no obvious difference between the image quality in either A and B groups or C and D groups(P>0.05).Based on the principle that low mAs resulted in low radiation dose and high kV lead to low effective dose,A group had the radiation dose lower than those of B group,and C group had the effective dose lower than those of D group. Conclusion SIEMENS Ysio DR has the photographing parameters regulated according to different sites and the amplification coefficient for enhancing image quality, which decreases X-ray radiation dose and effective dosage with the diagnosis results unaffected.

10.
Tianjin Medical Journal ; (12): 284-287, 2018.
Article in Chinese | WPRIM | ID: wpr-698025

ABSTRACT

Objective To investigate the clinical usefulness and safety of soluble dietary fiber (SDF) combined with probiotics in treating elderly patients with antibiotic associated diarrhea(AAD).Methods A total of 88 hospitalized elderly patients (≥65 years old) treated with antibiotics during August 2014 to August 2016 were included in this study. Patients were divided into control group(26 cases),probiotics group(30 cases)and combined group(32 cases).The control group was given maltodextrin as placebo intervention, the probiotics group was given the same dose of probiotics only, and the combined group was given SDF and probiotics treatment. The treatment was observed for 14 d. The incidence and the recurrence rate of ADD, the results of fecal culture and the incidence of adverse reactions were monitored in the three groups. Results The incidence of ADD was obviously lower in the combined group and the probiotics group than that of control group(6.25% vs.26.92%,6.67% vs.26.92%,P<0.05).The disease duration of diarrhea was shorter in the combined group than that of probiotic group[(3.53 ± 0.62)d vs.(4.39 ± 1.01)d,P<0.05],but no difference was found when compared with that of control group [(3.55 ± 0.65) d]. After 7-day intervention, Enterococcus counts was significantly deceased in probiotic group(P<0.05).There were no significant changes in Lactobacillus and Bifidobacterium counts in probiotic group after 14-day intervention. In the combined group, after 7-day intervention, Enterococcus counts decreased (P<0.05), and then started increasing until the 14-day(P<0.05).Lactobacillus and Bifidobacterium counts increased with the time passed (P<0.05).The incidence rates of total adverse reactions were 6.25%,10.00% and 23.07% for combined group,probiotics group and control group,and with no significant differences between them.Conclusion The probiotics combined with SDF has a good clinical effect on preventing the occurrence of antibiotic associated diarrhea and shortening the course of diarrhea in the elderly.It has high safety and it can regulate the intestinal flore with high safety.

11.
International Eye Science ; (12): 712-713, 2015.
Article in Chinese | WPRIM | ID: wpr-637239

ABSTRACT

AIM: To observe the changes in subfoveal choroidal thickness ( SFCT ) after intravitreal injections of ranibizumab ( IVR ) for macular edema secondary to retinal vein occlusion ( RVO) . METHODS:Thirty-six eyes of 36 patients with macular edema secondary to RVO) were treated with 0. 5mg IVR monthly for 3mo and received additional IVR as needed over the following 1a period. SFCT of the all eyes ( the affected eyes with RVO and unaffected fellow eyes ) was measured by enhanced depth imaging optical coherence tomography images before and after the IVR. RESULTS: The mean SFCT of the affected eyes with RVO decreased from 246. 7±115. 0μm at baseline to 220. 5±102.0μm at 1mo (P0. 05), 228. 6±127. 0μm at 6mo (P>0.05), 223.6±101.0μm at 12mo(P>0.05). There were statistically significant difference between affected eyes with RVO and unaffected fellow eyes. CONCLUSION: The SFCT is decreased after IVR for macular edema secondary to RVO. IVR seems to affect the hemorheology of the choroid.

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