Научная статья на тему 'Research on the effective fraction of Shaofuzhuyu Decoction through human primary end ometriosis cells'

Research on the effective fraction of Shaofuzhuyu Decoction through human primary end ometriosis cells Текст научной статьи по специальности «Фундаментальная медицина»

CC BY
50
7
i Надоели баннеры? Вы всегда можете отключить рекламу.
Ключевые слова
SHAOFUZHUYU DECOCTION / ENDOMETRIOSIS / PRIMARY CELL CULTURE

Аннотация научной статьи по фундаментальной медицине, автор научной работы — Wang Xu, Zangjin Qi, Sun Xiaolan, Zhao Chuang, Sun Ze

Endometrial primary cells as in vitro cells can better respond to endometrial physiological activity, the research of uterine-related diseases is of great significance.The endometrial cells were digest-ed by enzymatic digestion, and the endometrial stromal cells and epithelial cells were separated by dif-ferential centrifugation and sieve method. Study on the active fraction activity of Shaofuzhuyu Decoction.

i Надоели баннеры? Вы всегда можете отключить рекламу.
iНе можете найти то, что вам нужно? Попробуйте сервис подбора литературы.
i Надоели баннеры? Вы всегда можете отключить рекламу.

Текст научной работы на тему «Research on the effective fraction of Shaofuzhuyu Decoction through human primary end ometriosis cells»

of the traditional Chinese medicine Qingfeng rattan alkaloid extractive sinomenine in treatment of chronic nephritis l.Inhibit the expression of ICAM-1

Inthenormalphysiologicalstate,intercellularadhesionmoleculeICAM-1isalmostnoexpression,butwhentherenalinflam-matorydiseaseoravarietyofimmunereactionsleadtothebodyproducesvariouskindsofkidneydisease,ICAM-1expressed significantly in renal tubular epithelial cells, globular mesangial cells, renal tubular peripheral vascular endothelial cells.

2.Down-regulated TNF-a expression TNF-a is a small molecule protein mainly secreted by mononuclear - macrophages, the study found that excessive TNF-aexpression, will stimulate mononuclear cells and T lymphocyte to secrete hormones, promote proliferation of glomerular mesangial cell, increase sclerosis of glomerular. TNF-acan also affect the function of glomerular mesangial cells, induced by the formation of a large number of platelet-activating factor (PAF), stimulate mesangial cell proliferation, division. The sinomenine can reduce the expression of TNF-a and TNF-amRNA, improve the pathological changes of chronic nephritis, delay glomerular sclerosis.

3.Immunosuppressive effects Lymphocytes are an important component of the immune response of the body, in which the main constitute of lymphocytes T lymphocytes, can directly kill the target cells, auxiliary and inhibition of B cell antibodies and so on. The new member Th17 of T cell subgroup can promote the production of IL-17 factor, it can also stimulate the body to produce inflammatory response, leading to related autoimmune diseases.Studies have shown that, sinomenine novel derivatives 1032 can selectively inhibit the differentiation of Th17 cell, thereby reducing the inflammatory response of autoimmune encephalitis.

Conclusion In recent years, domestic and foreign scholars have shown that sinomenine has anti-inflammatory,immuno-suppressive,anti-tumor and other biological activity through a large number of animal experiments and clinical observation studies , for the treatment of chronic nephritis provides a theoretical and practical basis. However, the current market common sinomenine formulations are more single. The scope of application is also concentrated in the rheumatoid joint disease, while the sinomenine itself has the easy decomposition, short biological half-life characteristics, resulting in the useofdose is toolarge,seriouslyaffecting theconvenienceof clinical application of sinomenine.We hope thatwith the drug mechanism research of the sinomenine in-depth progress, the preparation process can be significantly improved, on the occasion the application of sinomenine-related agents in the treatment of chronic nephritis will have a broader prospect.

RESEARCH ON THE EFFECTIVE FRACTION OF SHAOFUZHUYU DECOCTION THROUGH HUMAN PRIMARY END

OMETRIOSIS CELLS

Wang xu , Zangjin qi, Sun xiaolan, Zhao chuang, Sun ze,Wu xiuhong*

National TCM Key Laboratory of Serum Pharmacochemistry, Laboratory of Metabolomics, Heilongjiang University of Chinese Medicine, Heping Road 24, Harbin 150040,China.*Correspondence: Xiuhong Wu; Email: [email protected]

Abstract Endometrial primary cells as in vitro cells can better respond to endometrial physiological activity, the research of uterine-related diseases is of great significance/The endometrial cells were digested by enzymatic digestion, and the endometrial stromal cells and epithelial cells were separated by differential centrifugation and sieve method. Study on the active fraction activity of Shaofuzhuyu Decoction.

Key words: ShaofuZhuyu Decoction, Endometriosis, Primary cell culture.

Objective Cultured human primary endometrial cells were used to observe the efficacy of different administra-tionfractions of ShaofuZhuyu Decoction.

Materials and methods No phenol red DMEM/F12 medium, Fetal bovine serum, Type IV collagenase, PBS,Penicillin-streptomycin solution,MTT,DMSO, the whole formula of Shaofuzhuyu Decoction,the alcohol sink fraction of shaofuzhuyu Decoction,Shaofuzhuyu Decoction 20% alcohol wash fraction,Shao-fuzhuyu Decoction 40% alcohol wash fraction,Shaofuzhuyu Decoction 60% alcohol wash fraction.

Patients from the Second Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine were treated with strict sterile conditions for the removal of ectopic endometrial tissue. Tissue was immediately placed in 10mL of 1% green-streptomycin-free phenol red DMEM/F12 medium and delivered to the laboratory within two hours.The tissue was retrieved with a Pasteur tube to wash the tissue twice and wash away the remaining blood clots. Cut the tissue into 1mm3 pieces with ophthalmic tweezers and add 10mL of type IV collagenase digestion solution to the carbon dioxide incubator for lhour and 20minutes. The digested tissue was pipetted into a 15mL centrifuge tube and centrifuged at 3minutes (500rpm). Supernatant mostly is endometrial stromal cells, precipitation mostly is endometrial epithelial cells. The upper fluid was aspirated into a 50mL centrifuge tube by a 40um cell sieve. The filtrate was endometrial stromal cells and the endometrial epithelium was on the filter.Spray 3mL of culture solution with a pipette to rinse the epithelial cells on the filter to a new 50 mL centrifuge tube.The initial centrifugation was suspended in 2mL of cell culture medium and the suspension was again filtered through a 40um cell.Stromal cells and epithelial cells collected in a 50mL centrifuge tube were transferred to a 15mL centrifuge tube, respectively. Endometrial stromal cells were centrifuged at 5min (1200rpm) and endometrial epithelial cells were centrifuged at 5min (1000rpm). Take the original endometrial stromal cells and epithelial cells according to cell count, diluted to 104-105, mixed cells to 200^L per hole in 96-well plate.After 48hours the replacement fluid removed the unattached tissue cells.72hours later, the five drugs

Амурский медицинский журнал №4 (20) 2017 69

were 20^g/mL, 2^g/mL, 0.2^g/mL concentration of each drug three holes.After 24 hours of administration, MTT dye was added to each well and the carbon dioxide incubator was added for 4 hours.discard the culture medium, plus 150^L of DMSO per well,the level of shock for 15 minutes, into the microplate reader at 492nm absorbance detection.

Results and discussion Shaofuzhuyu Decoction 40% alcohol wash fraction inhibit ectopic endometrial growth,and showed a gradient inhibition at the drug concentration.Most of the 60% alcohol wash fraction of Shaofuzhuyu Decoction inhibited cell growth, but no regularity was found at this concentration.lt can be presumed that the main components of the 40% alcohol wash fraction and the 60% alcohol wash fraction of the Shaofuzhuyu Decoction are the effective combination of isorhamnetin-3-O-neopalopyranoside and typhaneosidein the treatment of endometriosis ingredients.Ferulic acid and paeoniflorin in the 60% alcohol wash fraction may also be an active ingredient in the treatment of endometriosis.

Shaofuzhuyu decoction of complex ingredients, simple animal pharmacological experiments can't be a comprehensive response to the basis of drug substance.Through in vitro culture of human primary endometriosis cells, ShaofuzhuyuDecoction effective ingredients can be more accurate understanding/The lack of experimentation is that there is no validation of the interaction between the monomer or monomer components in the different extraction fractions, which is what we will do in the next part of the experiment.

References:

1Bo Hyon Yun,SeungJoo ChonYoung Sik Choi,etal.Pathophysiology of Endometriosis: Role of High Mobility Group Box-1 and TollLike Receptor 4 Developing Inflammation in Endometrium[J].Plos One, 2015,11(2):1-17.

2 Joseph C.Chen,Nadia R. Roan.lsolation and Culture of Human Endometrial Epithelial Cells and Stromal Fibroblast]. HHSPublic Access, 2015,5(20):1-10.

THE DEVELOPING OF THE PHARMACOLOGICAL ACTION OF HYDROXYTYROTOL

Wanmeng Zhang,Yongji Li*

(Research Institute of College of Pharmacy, Heilongjiang university of Chinese Medecine, Harbin, 150040, China)

Abstract: Hydroxytyrosol is a kind of polyphenol compounds exists in the olive,has good biological effects.This paper reviews the action of hydroxy tyrosol in anti-cancer,resistant microorganisms,antithrombotic,regulation of blood lipids and resist arteriosclerosis,prevention and control of hepatitis b resistance,macular degeneration,protect cartilage and anti osteoporosis and health care function and other aspects of pharmacological effects and mechanism.

Keywords: Hydroxytyrosol,Anticancer,antimicroorganism,antithrombus,Regulating,blood lipids and antiatherosclero-sis

Lilac leaves are dried leaves of lilacs. The study of lilacs in modern times has been more extensive, and its active substances include Hydroxy butyl alcohol . Hydroxytyrosolhas a strong antioxidant activity,is also a important chemical composition of lilac leaves. In recent years,the study found that the hydroxyl tyrosol has the pharmacological effects on anti-cancer,resistant microorganism,antithrombotic,regulation of blood lipids and resist arteriosclerosis,preven-tion and control of hepatitis b resistance,macular degeneration,protect cartilage and anti osteoporosis and health care function at home and abroad,so the development of drugs for treating diseases will have certain significance.

1 Anticancer Cancer is one of the major public health problems recognized in many countries.ln the United States,tumors are the only cause of death after cardiovascular disease.ln China,the death rate of cancer is increasing year by year.The excess of free radicals and reactive oxygen in the body destroys DNA and causes the gene to express an abnormal expression of the tumor.Hydroxy butyl alcohol with pyrocatechol structure,is a typical oxidation stress material and free radical scavenger,which provided a certain basis for the pharmacological effects of anti-tumor [1]. Studies have shown that hydroxytyrol has a inhibiting effect on multiple stages of tumor development.

2 Anti-pathogeny microorganism The anti-pathogeny microorganism of hydroxytyroxol is characterized by wide spectrum and high efficiency.lt is used as a food additive based on its antimicrobial advantage. HT not only has antimicrobial activity for harmful bacteria in the intestinal tract,but also has inhibitory effect on bacillus. Hydroxy butyl alcohol not only can be used as antibacterial, antiviral drugs,can also be used to strengthen the function of the body's defense,all reflect the HT development potential for disease resistance of original microbial drugs.

3 Antithrombus; Regulatingblood lipids and antiatherosclerosis HT inhibits theadhesion andactivityof platelets toprevent the formation of clots.The resistance of cardiovascular disease may be related to lower blood sugar and lipid oxide concen-tration,the datas show that the hydroxyl butyl alcohol can also be used for diabetes,high blood sugar and oxidative stress symptoms[2].The hydroxytyrotol protects the cells of the body from oxidation and prevents hardening of the arteries by inducing the expression of the activation of adenosine activation protein kinase and activating the active FOXO3a enzyme.

4 Other pharmacological effect ln addition to the above pharmacological effects,hydroxytyrotol also has the role of preventing macular degeneration,protecting cartilage and preventing osteoporosis and health care.And environmental toxicity and oxidation of fat granule and age factors such as prevention effects of the oxidative damage epithelial cells of the retina,especially suitable for the elderly to maintain eye health and high resolution,so as to improve the effect of vision.The cartilage is a dense connective tissue, such as osteoblasts or chondrocytes,which are buried in the extracellular matrix. lt is an important part of the skeleton.Hydroxy tyrosol played an important role

70 Амурский медицинский журнал №4 (20) 2017

i Надоели баннеры? Вы всегда можете отключить рекламу.