Investigation Of The Mechanism And The Effect Of Marrow Mesenchymal Stem Cell On The Mice With Systemic Lupus Erythematosus

Objective: To study whether the mesenchymal stem cells (MSC) infusion can reduce the autoimmune response of systemic lupus erythematosus (SLE) in mice, improving their immune injury, and to explore the mechanism.Methods: 1) separation of rat bone marrow MSC: separation of male SD rats (80g) femur, tibia, using whole bone marrow culture isolated, amplified, and passaged ,resuscitation MSC, and depicts the growth curve. 2)Animal groups: the female MRL / mp (fas) lpr mice were randomly divided into two groups(n=5), experimental group: Infuse MSC (0.3×106/0.15ml) in the 14 weeks of age By tail vein; control group: normal saline instead of MSC. 3)The following indicators were observed for 8 weeks:①General situation of mice were observed every day: eating, mental, hair, weight, survival time until death.②Urine was collected every 3 days, Urinary protein concentration was determined by bradrord method.③Blood was collected by cutting tails, 50ul every week, separate the serum, measured by radio immunoassay of serum anti-ds-DNA antibody concentration.④mice's kidney were cut for pathological biopsy , observed Pathological changes, and Checked immune complexes by immunohistochemistry also.⑤CD4 + CD25 + T cell level in peripheral blood, spleen, thymus was measured by flow cytometry CD4 + CD25 + T cell level.Results: 1)With the methord of culturing whole bone marrow,the cellls will cover the bottom by 85-95% after 7-10 days. Collect the cells after the grouth of 2,3-generation , the growth curve showing the primary to the three generations of MSC growth condition is better.2) Intravenous infusion, the mice were partial, total no adverse reaction.3) During the deservied 8 weeks:①two Groups of rats are all survived for 8 weeks .Their eating, hair, mental status, activity, body weight more or less are the same until death.②Before the experience,2 groups's Urine Protein density has no statistic difference(P <0.05).The test group's urinary protein density was prominent lower than the control group's .③The density of anti-ds-DNA antibody in test group was significantly lower than control group's(P <0.05).④HE staining showed renal pathology in control mice showed Mesenchymal nephritis were paramount; mouse kidney immune complex deposition showed punctate distribution in the test group, and its involving range is less than the control group (P <0.05).⑤the peripheral blood,thymus CD4 + CD25 + T cell level is higher than the proportion of experimental group, statistics treating show significant difference (P <0.05); spleen CD4 + CD25 + T cells in experimental group is slightly higher than control group , no statistical difference (P> 0.05).Conclusion: MSC is easy to grow and be separated in vitro,and there has no priminent untoward effect after infusion .After the infusion ,rats will improve the situtation of their autoimmune response injury. Its mechanism may reduce their own anti-ds-DNA antibody production and ameliorate their renal immunology pathology injury by improving the MRL/mo(fas)lpr small rats'CD4 + CD25 + T-cell level.