Study On Cell Apoptosis By Lentivus Perk And Construction Of PGRN^-/- Mice Arthritis Model Induced By Type Ⅱ Collagen

Objective To construct the protein kinase R-like endoplasmicreticulum kinase (Protein Kinase R-like ER kinase, PERK) lentiviralvectors and lentiviral particles (Lentivirus), explore the chondrocytesPERK lentiviral gene modified endoplasmic reticulum stress guideapoptosis; while building chicken collagen type Ⅱ induced PGRN-/-mouse model of arthritis. Methods Lentiviral vector systempWPT-GFP-PERK lentiviral packaging plasmid pMD2G, pSPAX2into293T cells were further formed PERK lentivirus packaging (LV-PERK)(see first part); cultured myoblast C2C12, with tunicamycin(tunicamycin, TM) within the endoplasmic reticulum stress inducers (ERSress), was observed in the ER Sress PERK state influence (see part II)on C2C12cell proliferation and apoptosis; and C3H10and ATDC5micromass culture cells, BMP2induced at different time points wereinfected LV-PERK, LV-GFP, to detect the effects (see part III) oncartilage differentiation process of apoptosis; chicken collagen type Ⅱ(COLII) and Freund’s complete adjuvant (FCA) by mixing equal volumes of the tail subcutaneous PGRN-/-mice,3weeks after the injection of thesame amount of the same COLII, observed PGRN-/-mice has occurredwith rheumatoid arthritis (see see Part III). Results Was successfullyconstructed and packaged lentiviral human PERK; under TMendoplasmic reticulum stress-induced state, PERK can accelerateapoptosis; in process ATDC5BMP2induced chondrocyte differentiationand C3H10, PERK can reduce the differentiation process apoptosis ofchondrocytes. Successful establishment of chicken collagen type Ⅱinduced PGRN-/-mouse model of arthritis. Conclusion endoplasmicreticulum stress state, PERK can accelerate apoptosis in chondrocytedifferentiation process, PERK can reduce apoptosis during differentiationof chondrocytes. To reveal the regulatory mechanism under conditions ofendoplasmic reticulum stress and cartilage cell differentiation PERKtheoretical basis for the mention. PGRN-/-mice established arthritismodel, provides a new model for arthritis research.