丙泊酚通过JAK激酶-信号转导及转录活化因子信号通路抑制类风湿关节炎大鼠炎症反应及滑膜细胞凋亡

Propofol inhibits inflammatory response and synovial cell apoptosis in rheumatoid arthritis mice through the JAK kinase-signal transducer and activator of transcription pathway

目的研究丙泊酚对类风湿关节炎(RA)大鼠炎症反应、滑膜细胞凋亡、踝关节组织病理学变化及JAK激酶(JAK)-信号转导及转录活化因子(STAT)信号通路的影响。方法于2018年7月至2019年7月,选取36只Wistar雄性大鼠,采用随机数字表法分为六组,每组6只,分别为对照组、模型组、甲氨蝶呤组、丙泊酚低、中、高剂量组。除对照组外,其余各组均使用牛Ⅱ型胶原诱导建立RA大鼠模型,造模完成后,丙泊酚低、中、高剂量组分别按照10 mg/kg、20 mg/kg、40 mg/kg腹腔注射给予大鼠丙泊酚,甲氨蝶呤组按照7.5 mg/kg腹腔注射给予大鼠甲氨蝶呤,对照组及模型组腹腔注射给予大鼠等量生理盐水,每日定时给药1次,连续给药28 d后,对大鼠进行关节炎评分,检测大鼠血清肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)及白细胞介素-1β(IL-1β)水平及脾脏、胸腺脏器指数,流式细胞术检测滑膜细胞凋亡率,苏木精-伊红染色对大鼠踝关节进行组织病理学检查,蛋白质印迹法检测各组大鼠滑膜细胞的JAK2、STAT3水平。结果连续给予RA大鼠不同剂量的丙泊酚或甲氨蝶呤28 d后,与模型组比较,丙泊酚低、中、高剂量组及甲氨蝶呤组RA大鼠关节炎评分[(6.76±1.43)、(4.37±0.87)、(2.68±0.26)、(2.51±0.64)比(12.68±0.67)]、体内TNF-α[(26.67±1.81)ng/L、(19.44±1.34)ng/L、(14.79±1.36)ng/L、(14.39±1.54)ng/L比(32.87±1.95)ng/L]、IL-6[(52.86±1.37)ng/L、(45.71±2.01)ng/L、(38.55±2.71)ng/L、(35.66±1.82)ng/L比(67.71±1.34)ng/L]及IL-1β[(43.29±1.47)ng/L、(38.64±1.44)ng/L、(18.72±1.26)ng/L、(18.04±2.41)ng/L比(58.89±1.73)ng/L]水平、脾脏指数[(33.27±2.84)%、(28.44±1.66)%、(18.73±1.18)%、(25.51±2.39)%比(38.64±4.68)%]及胸腺脏器指数[(12.32±1.03)%、(10.67±0.81)%、(9.42±0.75)%、(10.11±0.97)%比(14.62±1.37)%]、滑膜细胞凋亡率[(31.71±1.91)%、(24.84±1.91)%、(16.33±2.32)%、(15.62±2.52)%比(41.85±3.97)%]均显著降低(P<0.05),组织病理学检查结果表明,丙泊酚各剂量组及甲氨蝶呤组RA大鼠踝关节病理组织学得到显著改善,蛋白质印迹法结果表明,与模型组比较,丙泊酚各剂量组及甲氨蝶呤组大鼠组织JAK2水平[(0.76±0.04)、(0.59±0.04)、(0.42±0.04)、(0.41±0.05)比(1.19±0.16)]、STAT3水平[(0.73±0.07)、(0.55±0.05)、(0.35±0.08)、(0.37±0.06)比(0.99±0.05)]均显著降低(P<0.05),且各项指标变化均呈现出对丙泊酚的剂量依赖性。结论丙泊酚能够抑制RA大鼠体内炎症反应及滑膜细胞凋亡,其作用机制可能与调节RA大鼠体内JAK-STAT信号通路有关。

Objective To study the effects of propofol on inflammatory response,synovial cell apoptosis,histopathological changes of ankle joint and JAK kinase(JAK)-signal transducer and activator of transcription(STAT)signaling pathway in rheumatoid arthritis mice.Methods From July 2018 to July 2019,thirty-six male Wistar mice were randomly assigned into 6 groups with 6 mice in each group by random number table:control group,model group,methotrexate group,low-,middle-and high-dose propofol groups.Except for the control group,all the other groups were induced by bovine typeⅡcollagen to establish rheumatoid arthritis mouse model.After the establishment of the model,the low-,middle-and high-dose groups of propofol were given intraperitoneal injection of propofol(10 mg/kg,20 mg/kg and 40mg/kg respectively),the methotrexate group was given intraperitoneal injection of methotrexate(7.5 mg/kg),and the control group and model group were intraperitoneally injected with the same amount of normal saline.The medicine was given once a day for the respective group.After 28 days of administration,the arthritis score was measured,the levels of TNF-α,IL-6 and IL-1βin serum,the organ index of spleen and thymus,the apoptosis rate of synovial cells were detected by flow cytometry,and the histopathological examination of ankle joint was made by hematoxylin-eosin staining,Western blotting was performed to detect the levels of JAK2 and STAT3 in synovial cells of the mice in each group.Results After 28 days of continuous administration of different doses of propofol or methotrexate in rheumatoid arthritis mice,compared with the model group,the arthritis score[(6.76±1.43),(4.37±0.87),(2.68±0.26),(2.51±0.64)vs.(12.68±0.67)],the levels of TNF-α[(26.67±1.81)ng/L,(19.44±1.34)ng/L,(14.79±1.36)ng/L,(14.39±1.54)ng/L vs.(32.87±1.95)ng/L],IL-6[(52.86±1.37)ng/L,(45.71±2.01)ng/L,(38.55±2.71)ng/L,(35.66±1.82)ng/L vs.(67.71±1.34)ng/L]and IL1β[(43.29±1.47)ng/L,(38.64±1.44)ng/L,(18.72±1.26)ng/L,(18.04±2.41)ng/L vs.(58.89±1.73)ng/L],splenic index[(33.27±2.84)%,(28.44±1.66)%,(18.73±1.18)%,(25.51±2.39)%vs.(38.64±4.68)%]and thymus index[(12.32±1.03)%,(10.67±0.81)%,(9.42±0.75)%,(10.11±0.97)%vs.(14.62±1.37)%]and the apoptosis rate[(31.71±1.91)%,(24.84±1.91)%,(16.33±2.32)%,(15.62±2.52)%vs.(41.85±3.97)%]of synovial cells in low-,middle-and high-dose propofol groups and methotrexate group were significantly lower(P<0.05).The histopathological test results showed that the histopathological conditions of ankle joint of rheumatoid arthritis mice in propofol groups and methotrexate group were significantly improved.Western blotting results showed that compared with the model group,the levels of JAK2[(0.76±0.04),(0.59±0.04),(0.42±0.04),(0.41±0.05)vs.(1.19±0.16)]and STAT3[(0.73±0.07),(0.55±0.05),(0.35±0.08),(0.37±0.06)vs.(0.99±0.05)]in tissues of mice in propofol groups and methotrexate group were significantly decreased(P<0.05),and the changes in all indexes were dose-dependent on propofol.Conclusion Propofol can inhibit the inflammatory reaction and synovial cell apoptosis in rheumatoid arthritis mice,whose mechanism may be related to the regulation of JAK-STAT signal pathway in rheumatoid arthritis mice.