Evaluation of Animal Model of Rheumatoid Arthritis Induced by Mycobacterium tuberculosis from Inflammation， Apoptosis and Autophagy
10.13422/j.cnki.syfjx.20221941
- VernacularTitle:从炎症、凋亡与自噬评价Mtb诱导类风湿关节炎风湿热痹动物模型
- Author:
Liping FU
1
;
Jiayu LI
1
;
Qijin LU
1
;
Yisi CAI
1
;
Yuxue ZHANG
1
;
Lixia YUAN
1
;
Gang LIU
1
Author Information
1. Nanfang Hospital， School of Traditional Chinese Medicine， Traditional Chinese Pharmacological Laboratory， School of Laboratory Medicine and Biotechnology， Southern Medical University， Guangzhou 510515， China
- Publication Type:Journal Article
- Keywords:
rheumatoid arthritis;
wind-damp-heat arthralgia;
disease-syndrome combined model;
apoptosis;
autophagy
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(23):44-52
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo evaluate the animal model of rheumatoid arthritis （RA） induced by inactivated Mycobacterium tuberculosis （Mtb） from inflammation， apoptosis and autophagy， compare the rat RA model of wind-damp-heat arthralgia （FSR） induced by Mtb with the rat RA model of adjuvant arthritis （AA）， and provide experimental evidence for improving the disease-syndrome combined model and developing drugs for the prevention and treatment of RA. MethodThirty 6-week-old SD rats were randomly assigned into 3 groups： a normal group， an AA group and a FSR group， with 10 rats in each group. The RA rat model was established by injection of Mtb suspension （1 mg/rat）， and the rats in the normal group were injected with the same volume of normal saline. The invention in the FSR group lasted for 16 days. The general conditions， body weight， spleen index， swelling of ankle joints， arthritis index （AI）， and the symptoms of arthritis in the hind feet of the rats in each group were observed and measured. The expression levels of serum interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） were detected by enzyme-linked immunosorbent assay （ELISA）. The expressions of IL-1β， TNF-α， apoptosis-related protein B cell lymphoma-2 （Bcl-2） and Bcl-2 associated X protein （Bax）， autophagy-related protein 1 light chain 3 （LC3）， autophagy key molecule yeast Atg6 （Beclin1），and p62 mRNA expression levels were detected by Real-time polymerase chain reaction （Real-time PCR）. The protein expression levels of Bcl-2， Bax， LC3， Beclin1 and p62 were detected by Western blot. ResultCompared with those of the normal group， the rats of AA and FSR groups grew slowly and presented dull hair， soft or loose stool， slow movement， swelling of spleen， redness and swelling of ankle joints， increased AI， and histopathologic changes in the synovium and ankle joints. Moreover， the modeling elevated the levels of IL-1β and TNF-α in the serum， up-regulated the mRNA levels of IL-1β and TNF-α in the synovium， up-regulated the mRNA and protein levels of Bcl2， LC3， Beclin1， and down-regulated the mRNA and protein levels of Bax and p62. Compared with the AA group， the FSR group showed severe symptoms， slowly increased body weight （P<0.01）， early appearance of obvious redness and swelling of ankle joints， increased AI （P<0.05）， increased spleen index （P<0.05）， and obvious pathologic changes of synovial tissue and ankle joints， the inflammatory infiltration in the synovial tissue， and the damage of joint structure. Moreover， the FSR group had higher expression levels of IL-1β， TNF-α， Bcl-2， LC3， and Beclin1 （P<0.05） and lower expression levels of Bax and p62 than the AA group （P<0.05）. ConclusionThe animal model of RA （syndrome of wind-damp-heat arthralgia） can be induced successfully with the symptoms consistent with clinical manifestations of RA. The FSR group has lower apoptosis level and higher autophagy level than the AA group， which indicates that the FSR group is more appropriate for the research on the RA with the syndrome of wind-damp-heat arthralgia.
