The Mechanism Of SPID Rat By Treating With Jialian Pelvic Capsule From The Correlation Between HMGB-1 And TLR2/4-NF-κB Pathway

Objective: This study is to observe the anti-inflammatory effect of sequelae of pelvic inflammatory disease,and explore the possible mechanism from the interaction of high mobility group box 1 protein with TLRs/NF-κB signaling pathway.Methods: Among the 90 female unpregnant Sprague Dawley rats,12 rats were randomly selected as the normal control and 12 rats were chosen as the sham-operated group,and the remaining were used to establish the model of sequelae of pelvic inflammatory disease induced by Phenol Plasma.After the successful establishment of the model,the rats with SPID were randomly divided into 5 groups including:SPID group,L+SPID,M+SPID,H+SPID and A+SPID group,12 in each group.To the L+SPID,M+SPID,H+SPID,the rats were given 18.75 mL/kg/d gavage of Jialian pelvic capsule suspension with concentrations of 20,40,80 mg/mL,respectively.The A+SPID group was given 6.25 mL/kg/d Fukeqianjin capsule(concentration: 40 mg/mL).The others were given equal volume of distilled water.The behavioral of rats,the gross situation of pelvic cavity and uterus in each group were observed visually and the pathological morphology of uterus was observed under light microscope.ELISA was used to detect the expression of HMGB-1,TNF-α,IL-6,IL-10.Immunohistochemical was used to observe the average optical density of TLR2,TLR4,MyD88,NF-κB/p65;and western blot was used to detect the protein levels of TLR2,TLR4,MyD88,phosphorylation of NF-κB/p65.Results:1 SPID model verification: rat pelvic congestion,effusion or accumulation of pus,tissue adhesion;uterine texture is hard,irregular morphology,uterine cavity dilatation or adhesion narrow;microscopic examination the uterine structure disorder,cavity wall structure destruction,intima hyperemia edema,epithelial cell degeneration and necrosis,gland expansion or disappearance,a large number of inflammatory cell infiltration.2General condition of rats: rats in Con and Sham group were in good condition with normal physiological growth.SPID rats have sparse hair,irritability,increased corneal secretion,arch back,curled up,piled up,reduced diet,urine blood,and unformed stool.The physiological functions of rats in each treatment group were improved after the corresponding drug intervention,and rats in the H+SPID group were the most obvious.3 Morphological changes:Pelvic manifestations: the pelvic cavity of rats in con and sham group is well organized.The pelvic connective tissue of rats in SPID group was thicker than before.treatment groups pelvic no mass,congestion,tissue adhesion and other inflammatory manifestations alleviated;especially in the H+SPID.Pelvic adhesion degree score: compared with Con and Sham group,the degree in SPID group was significantly aggravated(P<0.001),while in each treatment group it was reduced(P<0.05),and H+SPID group showed the best effect(P<0.001).Uterine morphology: in Con and Sham group,the uterine cavity was unobstructed;The uterine inflammation in each intervention group was reducedand it was the most obvious in the H+SPID group.Uterine pathological morphology: The uterine structure of Con and Sham group was with clear structure and no obvious inflammatory cell infiltration;the uterine structure of H+SPID group was basically clear,the uterine cavity wall was intact,a few inflammatory cells were infiltrated,the glands were basically intact,and the smooth muscle cells were arranged neatly;followed by that was the M+SPID group,then the L+SPID and A+SPAD group.Uterine pathology score: the score of rats in SPID was significantly higher than that in con group and sham group(P<0.001);but lower than each treatment group(P<0.05).4.Rat uterine tissue HMGB-1,IL-6,TNF-α and IL-10 levels: compared with Con and Sham group,SPID HMGB-1,IL-6,TNF-α levels increased,while IL-10 level decreased,the difference was statistically significant(P<0.001);HMGB-1,IL-6,TNF-αdecreased and IL-10 level increased in each intervention group,the difference was statistically significant(P<0.05),especially in the H + SPID group(P<0.001).5 TLR/4-NF-κB pathway-related protein expression in rat uterus:compared with Con group and Sham group,the expression of TLR2,TLR4,MyD88,NF-κB p65,p-NF-κB p65 in SPID was significantly increased.The difference was statistically significant(P<0.001)while the above protein expression was reduced in each treatment group,and the most significant in the H + SPID group.Conclusion: 1 Rats with SPID caused by Phenol Plasma was confirmed to be reliable.2 HMGB-1,TLR2,TLR4,MyD88-NF-κB may be involved in the pathogenesis of SPID rats.3 Jialian pelvic capsule can effectively reduce theinflammatory response of SPID rats,repair uterine inflammatory pathological injury,and the curative effect is dose-dependent.4 The effect of Jialian pelvic capsule on SPID rats may be to reduce the inflammatory response by reducing the release of endogenous risk factor HMGB-1,inhibiting the activation of TLR2/4-MyD88-NF-κB signaling pathway,lowering the levels of inflammatory factors IL-6 and TNF-α,and promoting the release of inflammatory suppressor IL-10.5 The efficacy of Jialian pelvic capsule on the anti-inflammatory effect of SPID rats is equivalent to Fukeqianjin capsule,which can be popularized.