补肾活血方对强直性脊柱炎小鼠的保护作用及对SIRT1表达的影响OA

Objective It is to observe the anti-inflammatory effect and its mechanism of prescription for tonifying kidney and activating blood(PTKAB)in the treatment of ankylosing spondylitis(AS)in mice.Methods ①Forty DBA/1 male mice were randomly divided into model group,low dose PTKAB group,high dose PTKAB group and celecoxib group,an-other 10 C57BL/6 mice were selected as the control group.The low dose PTKAB group and high dose PTKAB group were given PTKAB 12 g/kg,24 g/kg via gavage,respectively,while the model group and control group were given the same a-mount of normal saline via gavage,all once daily,continuously treated for 4 weeks.The degree score of joint swelling was evaluated,the histopathological morphology of the achilles tendon tissue was observed by HE staining,the serum levels of interleukin(IL)-6,IL-1β,IL-23,IL-17αand tumor necrosis factor-α(TNF-α)were measured by ELISA,and the protein expressions of silent information regulator 1(SITR1),CD86,and inducible nitric oxide synthase(iNOS)in achilles ten-don tissue were detected by Western blot.②THP-1 cells were taken and cultured in vitro,4 groups were set up in the ex-periment.The control group was cultured with 20 nmol/L PMA,the lipopolysaccharide(LPS)group was cultured with 20 nmol/L PMA+100 ng/mL LPS,the PTKAB group was cultured with 20 nmol/L PMA+100 ng/mL LPS+medicated ser-um containing 10%PTKAB,the PTKAB+si-SIRT1 group was cultured with si-SIRT1 plasmid transfection+20 nmol/L PMA+100 ng/mL LPS+medicated serum containing10%PTKAB.After24 hours of treatment,the protein expressions of SIRT1 and iNOS in the cells were detected by Western blot,and the expression of iNOS in the cells was detected by im-munofluorescence.Results ①Compared with the model group,the joint swelling scores and serum levels of IL-6,IL-1β,IL-23,IL-17α and TNF-α of mice in the medicated groups were significantly reduced(all P<0.05),and the inflammato-ry cell infiltration in achilles tendon tissue was significantly alleviated;the relative protein expressions of SITR1 in achilles tendon tissue were significantly increased(all P<0.05),while the relative protein expressions of CD86 and iNOS in achil-les tendon tissue were significantly decreased in the low dose and high dose groups of PTKAB(all P<0.05).②Compared with the blank group,the relative protein expression of SITR1 was significantly reduced in the LPS group(P<0.05),while the relative protein expression of iNOS and the relative fluorescence intensity of iNOS expression were both significant-ly increased(both P<0.05).Compared with the LPS group,the relative protein expression of SITR1 was significantly in-creased in the PTKAB group(P<0.05),while the relative protein expression of iNOS and the relative fluorescence inten-sity of iNOS expression were both significantly decreased(both P<0.05).Compared with the PTKAB group,the relative protein expression of SITR1 was significantly reduced in the PTKAB+si-SIRT1 group(P<0.05),while the relative pro-tein expression of iNOS and the relative fluorescence intensity of iNOS expression were both significantly increased(both P<0.05).Conclusion PTKAB can inhibit inflammation through regulation of SITR1-mediated polarization of macrophage M1,thus to improve joint swelling and slow down the progression of AS.