Therapeutic mechanism of Psammosilene tunicoides extract on rheumatoid arthritis based on NLRP3 inflammasome / 中国中药杂志

In this study, we investigated the mechanism of crude extract of Psammosilene tunicoides(CEPT) in the treatment of rheumatoid arthritis(RA) based on the Nod-like receptor protein 3(NLRP3) inflammasome. The collagen-induced arthritis(CIA) mouse model was established. On day 32 after the primary immunization, according to the arthritis score, the mice were randomly divided into model group, positive control(methotrexate) group, low-and high-dose CEPT groups, and normal group, with 10 mice in each group. According to the administration dose of each group, the mice were continuously administered for 21 days. Every four days during the administration, the paw edema degree, arthritis score, and spleen index of the mice were measured; histopathological examination was performed for the ankles of the mice; the contents of IL-1β and IL-18 in the serum were determined; the protein expression levels of NLRP3, caspase-1, and apoptosis-associated speck-like protein containing a CARD(ASC), as well as the mRNA expression levels of NLRP3 and caspase-1 in the ankle joints of the mice were detected. The results showed that compared with those in the model group, the mice in the positive control group and CEPT groups had significantly decreased the contents of IL-1β and IL-18 in the serum and spleen index(P<0.01), significantly lowered arthritis score and degree of paw edema(P<0.01), alleviated arthritic infiltration of the knee, and down-regulated protein and mRNA levels of NLRP3, ASC, and caspase-1 in the ankle joint(P<0.01). These results suggest that P. tunicoides may reduce the paw edema and arthritis score and alleviate the inflammatory response in CIA mice by inhibiting the expression of NLRP3. This study provides a basis for the study of immune regulation of P. tunicoides in RA.

Comparison of Improvement Effects and Its Mechanism of Different Doses of Total Alkaloids from Aconitum racemulosum on CIA Model Rats Based on Information Entropy Theory / 中国药房

OBJECTIVE： To investigate the effects of different doses of total alkaloids from Aconitum racemulosum （ARTA） on serum inflammation factors and FOS protein expression in synovial tissue of joint in collagen-induced arthritis （CIA） model rats， and to investigate its potential mechanism of anti-rheumatoid arthritis （RA）. METHODS： Male SD rats were randomly divided into blank group， model group， positive group （Compound dexamethasone acetate ointment， 0.2 g/kg）， ARTA low-dose， medium-dose and high-dose groups （56.26， 112.50， 225.00 mg/kg， by the weight of ARTA in the extract）， with 10 rats in each group. Except for blank group， other groups were given subcutaneous injection of Bovine collagen Ⅱ emulsified with incomplete Freund’s adjuvant into the left foot to establish CIA model； the left foot were smeared with relevant medicine from the day of modeling. Blank group and model group were smeared with constant volume of 65％ ethanol， 3 times a day， for consecutive 28 days. On the 7th， 14th， 21st and 28th day of administration， the thickness of left hind toe was measured with vernier caliper， and the degree of foot swelling was calculated. The serum contents of IL-1β， IL-6 and TNF-α in rats were measured by ELISA after last administration. The expression of FOS protein in synovial tissue was determined by immunohistochemical method [expressed by HIS]. The comprehensive score was conculated by entropy weight method. Effects of each dosage on above indexes of CIA model rats were evaluated with the comprehensive score. RESULTS： Compared with blank group， the degree of foot swelling， serum content of inflammatory factors and HIS value were increased significantly in model group （P＜0.05）. Compared with model group， the degree of foot swelling in each administration group， serum contents of IL-1β， IL-6 and TNF-α， HIS in positive group and ARTA high-dose group， serum contents of IL-6 and TNF-α in ARTA medium-dose group as well as serum content of TNF-α in ARTA low-dose group were decreased significantly（P＜0.05）. Comprehensive score of above indicators were 0.37（positive group）， 0.31（ARTA high-dose group）， 0.23（ARTA medium-dose group） and 0.09（ARTA low-dose group）. CONCLUSIONS： ARTA can improve CIA model rats， and the effect tends to increase with the increase of dose. Above effect may be associated with reducing serum content of inflammatory factors and inhibiting the expression of FOS protein in synovial tissue.

The expression of TIPE2 and its role in the pathogenesis of rheumatoid arthritis / 中华微生物学和免疫学杂志

Objective To investigate the expression of TIPE2 in splenic lymphocytes derived from the mouse model of cattle collagen Ⅱ-induced rheumatoid arthritis (CIA) and from peripheral blood of patients with rheumatoid arthritis (RA) for its role in the pathogenesis of RA.Methods The CIA mouse model was established by immunization of DBA-1/J mice with cattle collagen Ⅱ.Peripheral blood lymphocytes were collected from RA patients and healthy donors.The TIPE2 mRNA and protein expression in splenocytes of CIA and control mice were measured by semi-quantitative RT-PCR and Western blot,respectively.In addition,fluorescence quantitative RT-PCR and Western blot were used to determine the TIPE2 mRNA and protein expression in peripheral blood lymphocytes derived from patients with RA and healthy donors.Results Both mRNA and protein expression of TIPE2 were higher in splenocytes of CIA mice and lymphocytes of peripheral blood from RA patients compared with corresponding controls.Furthermore,the mRNA and protein expression of TIPE2 increased significantly in patients with active RA.TIPE2 expression were positively correlated with DAS28 scores (P＜0.001).Conclusion Our results suggest that TIPE2 plays a role in the RA pathogenesis.The level of TIPE2 may also indicate the severity of rheumatoid arthritis.

Study on establishing type Ⅱ collagen induced arthritis model in rats / 中国免疫学杂志

Objective:Research on establishing a better experimental arthritis rat model of cold-dampness arthraligia on rheumatoid arthritis(RA) research.Methods:In order to induce arthritis,the rat's feet were injected with different concentrations of type Ⅱ collagen mixed with Freund's adjuvant;measure the feet swelling degrees of rats,then calculated swelling rate;detect the level of serum sialic acid(SA);observ the synovial histopathology and ultrastructural of rats.Results:The rat joints swell of the group with low concentration of type Ⅱ collagen and completed Freund's adjuvant was rapid and severe,while that of the group with high concentration of type Ⅱ collagen and incompleted Freund's adjuvant was moderate and lasting.The level of serum SA of model groups was higher than the normal group.The histopathology changes of rat's synovium conform to RA.Conclusion:The group injected with high concentration of type Ⅱ collagen and incompleted Freund's adjuvant is the ideal RA model.It is belonged to the symptom of cold-dampness arthralgia,compared with this model,the AA approaches the symptom of heat-dampness arthralgia.