Jilin Province Great Great Pharmaceutical Co. LTD

Jilin Province Broadwell Pharmaceutical Co., LTD. Welcome you!

Service hotline:+86-431-85639055

博大伟业
imgboxbg

HEALTH SCIENCE

Your current location:
Homepage
/
/
/
Polyprezinc, a new way to protect gastric mucosa——inhibition of caspase-3 activity

Polyprezinc, a new way to protect gastric mucosa——inhibition of caspase-3 activity

  • Categories:Stomach healthy
  • Author:
  • Origin:
  • Time of issue:2020-12-03
  • Views:0

(Summary description)

Polyprezinc, a new way to protect gastric mucosa——inhibition of caspase-3 activity

(Summary description)

  • Categories:Stomach healthy
  • Author:
  • Origin:
  • Time of issue:2020-12-03
  • Views:0
Information

  Indomethacin (IND) is a non-steroidal anti-inflammatory drug (NSAID). Although non-steroidal anti-inflammatory drugs are used clinically to treat inflammatory diseases, they often cause side effects such as gastric mucosal damage. Studies have found Indomethacin can induce the release of cytochrome c, the activation of caspase-3 (caspase-3) and the apoptosis of rat gastric mucosal cells.

  Polyprezinc (PZ) is a chelate compound composed of zinc and L-carnosine, and is widely used clinically for gastric ulcers. Studies have shown that polyprezinc can effectively prevent gastric mucosal damage. Such protective effects are mainly attributed to its antioxidant properties, membrane stability and mucus stimulation. Therefore, we are very curious about the protective effect of polyprezinc on indomethacin-induced gastric mucosal cell apoptosis?

  Materials and Method

  1.1 Cell culture

  Rat gastric epithelial cell line RGM1 was cultured in a mixed medium at 37°C and 5% CO2 humidified environment.

  1.2 Cell pretreatment

  RGM1 cells were grown to near confluence, and the cells were immersed in serum-free DMEM/F12 medium. To determine the concentration of indomethacin that induces apoptosis of RGM1 cells, cells were exposed to 0-1000 μM IND for 0, 1, 3, 6, 12, and 24 hours at 37°C, 5% CO2/95% air. 30min before administration, PZ, ZnSO4, L-carnosine, and a mixture of ZnSO4 and L-carnosine were added to the medium.

  1.3 Fluorescence analysis of cell apoptosis

  Cell apoptosis was analyzed by flow cytometry, resuspended with trypsin digestion, washed twice with PBS, fixed with 70% ethanol, centrifuged for 5 min, washed once with PBS, suspended in citrate phosphate buffer, and at room temperature Leave it for at least 30 minutes. After washing with PBS, it was treated with 0.1 mg/mL RNase A at 37°C for 30 min, and stained with 50 ug/mL PI. Analyze 10,000 cells with an Epics cell fluorometer. Apoptotic cells showed a subduplex DNA peak (sub-G1), repeated at least 3 times.

  1.4 Caspase activity determination

  The activities of caspase-1 and caspase-3 proteins are collectively referred to below as YVADase and DEVDase, respectively. During incubation at 37°C for 60 minutes, YVADase and DEVDase are determined by the cleavage of fluorescent peptide substrates AC-YVAD-AMC and AC-DEVD-AMC active.

  1.5 Western blot

  The protein of each S-100 cytosol was quantified by Bradford method. Using gel electrophoresis separation method, using 12% polyacrylamide gel and polyvinylidene fluoride membrane, 10-20μg protein can be separated from each S-100 cytosol. 1.6 Determination of cell zinc content Use zinc-specific fluorescent dye zinquin ester to measure cell zinc content at 370nm and 490nm by fluorescence spectroscopy.

  1.7 Statistical analysis

  The result is expressed as the mean standard error of the mean (S.E.M.). The data shown is the average of at least three independent experiments. One-way analysis of variance was used for statistical analysis, and then Scheffe’s test was performed afterwards to determine the significant difference between the treatment groups. P<0.05 has a significant difference.

  Results and analysis

  2.1 IND induces apoptosis and DEVDase activation

  After RGMI cells were pretreated with indomethacin (500μM) for 3 hours, the cells began to undergo apoptosis. As time passed, the number of apoptosis continued to increase. After 12 hours, the number of apoptosis became stable; the activity of DEVDase increased with time. It continued to strengthen and reached the maximum after 12h. The activity of DEVDase in the control group was not affected, and the activity of YVADase remained unchanged.

  2.2 IND induces apoptosis and DEVDase activation in a dose-dependent manner

  Indomethacin concentration> 100μM, the induced apoptosis rate and indomethacin dose increased in a dose-dependent manner. When the concentration of indomethacin was less than or equal to 500μM, the DEVDase activity increased with the increase of the concentration, but after 1000μM indomethacin pretreatment, the DEVDase activity of the cells decreased significantly. This may be caused by cell necrosis, and YVADase activity remains unchanged.

  2.3 Polyprezinc significantly reduces IND-induced cell apoptosis and DEVDase activity

  After pretreatment with polyprezinc (25μM and 50μM), the number of apoptotic cells decreased significantly to 42% and 16% of the total cells, respectively. Polyprezinc pretreatment can also inhibit indomethacin-mediated DEVDase activity in a dose-dependent manner, and its DEVDase activity was significantly reduced by 63% and 92%, respectively.

  2.4 The effect of polyprezinc on the release of cytochrome c during IND-induced apoptosis

  "The release of cytochrome c from mitochondria to cytoplasm is the key to activate DEVDase during apoptosis. Polyprezinc does not affect cytochrome c efflux during indomethacin-induced apoptosis (Figure 2C), indicating that the molecular target of polyprezinc is located downstream of cytochrome c, and DEVDase is induced by indomethacin The upstream of apoptosis is activated.

  2.5 The effect of polyprezinc and its components on cell apoptosis and DEVDase activation

  As shown in Figure 3A, ZnSO4 (50μM) reduced the apoptosis rate induced by indomethacin by 69%. In contrast, the same concentration of L-carnosine could not inhibit cell apoptosis. The inhibitory effect of indomethacin-induced apoptosis is equivalent to that of polyprezinc. Whether the cells were pretreated with polyprezinc, ZnSO4 or a mixture of ZnSO4 and L-carnosine, the increase in DEVDase activity was completely inhibited, but L-carnosine pretreatment had no effect on DEVDase activity.

  2.6 The effect of polyprezinc and its components on intracellular zinc content

  The zinc content of cells pretreated with polyprezinc, ZnSO4, and a mixture of ZnSO4 and L-carnosine increased by 7.5 times, while the zinc content of cells pretreated with L-carnosine did not change.

  Discuss

  It can be seen that taking a non-steroidal anti-inflammatory drug-indomethacin (IND) can induce gastric mucosal cell apoptosis and caspase-3 activation, leading to gastric mucosal damage; polyprezinc treatment significantly reduces IND-induced cells Apoptosis and DEVDase activity protect the gastric mucosa from damage in a dose-dependent manner.

  Polyprezinc is a chelate composed of zinc and L-carnosine. Studies have shown that L-carnosine itself does not affect IND-induced apoptosis and caspase-3 activity, but its component zinc inhibits apoptosis in a series of The role and caspase-3 activation play a vital role.

  It is well known that the protective effect of polyprezinc on the gastric mucosa is achieved through its antioxidant effect, and this study fully shows that the mechanism of polyprezinc inhibiting apoptosis of gastric mucosal cells may be achieved through a new way-inhibition Caspase-3 activity. Boda Weiye Ruilaisheng (Polyprezinc Granules) can be used to treat gastric mucosal damage caused by long-term use of non-steroidal anti-inflammatory drugs (indomethacin).

Scan the QR code to read on your phone

Jilin Province Great Great Pharmaceutical Co. LTD

Service hotline

+86-431-81158731

CONTACT US

Jilin Province Great Great Pharmaceutical Co. LTD

Jilin Province Great Great Pharmaceutical Co. LTD
Jilin Province Great Great Pharmaceutical Co. LTD
Jilin Province Great Great Pharmaceutical Co. LTD
Jilin Province Great Great Pharmaceutical Co. LTD
Jilin Province Great Great Pharmaceutical Co. LTD
Jilin Province Great Great Pharmaceutical Co. LTD

Liaoyuan Production Base: No. 158 Fortune Road, Liaoyuan Economic Development Zone, Jilin Province

Tel:+86-437-6998023

Changchun R&D Center: No. 3786 Juye Street, Jingyue Development Zone, Changchun City, Jilin Province
Tel:+86-431-81158731(Marketing center)

Tel:+86-431-81158756(Research and development center)

Beijing Office: Room 901, Building F, Kaixuan City, 170 Beiyuan Road, Chaoyang District, Beijing
Tel:+86-10-58236233

Medication consultation, feedback on medication adverse reaction, and user complaint telephone:0437-5029815 

NO PUBLIC

Jilin Province Great Great Pharmaceutical Co. LTD

Scan and follow the official official account

Copyright © Jilin Province Great Great Pharmaceutical Co. LTD        吉ICP备20002630号

Website construction:300.cn Changchun   management     

Jilin Province Great Great Pharmaceutical Co. LTD