Effect Of Hydroxysafflor Yellow A On Airway Inflammation Via AMPK/NF-κB/NLRP3 In Asthmatic Mice

Objective:To study the effect of hydroxysaffloryellow A(HSYA)on airway inflammation in asthmatic model mice and its mechanismMethods:Forty female Balb/c mice were randomly divided into five groups: normal model group(control group),asthma model group(OVA group),HSYA low-dose group(HSYA-L)and HSYA high-dose group(HSYA-H),and dexamethasone positive control group(DEX group).A mouse asthma model induced by OVA was established.In addition to the normal control group,the OVA group,the HSYA-L HSYA-H group,and the dexamethasone-positive control group were given 200 μl of sensitized suspension per normal control group replaced with the same volume of saline on days 1,7,and 14,respectively,and each group was administered by intraperitoneal injection.From day 18,they were treated with intraperitoneal injection of hsya 200 mg/kg and hsya 100 mg/kg and dexamethasone 0.5 mg/kg,respectively,and the normal control group was replaced with normal saline for 7 consecutive days.Mice in each group were challenged from day 21,10 m L of 1% OVA was nebulized through a nebulizer,and the control group was replaced with 10 ml of normal saline for 30 min once daily for 3 days(challenge was started 1 hour after administration on days 21 –24).HSYA-L and HSYA-H mice were treated with the corresponding doses of drugs 30 min before each challenge.They were sacrificed 24 hours after challenge.The BALF,left lung tissue,and right lung tissue of the mice were taken and stored in the corresponding appropriate place for experimental use.HE and PAS staining were performed on the left lungs taken;Diff-quik staining was used to count the differential cells in bronchoalveolar lavage fluid(BALF);ELISA was used to detect the contents of interleukin-4(IL-4),IL-5,IL-13,and interferon-γ(IFN-γ)in BALF of mice;the expression of NF-κBp65 in lung tissue was observed by immunohistochemistry;and Western blot were used to detect protein expression changes of AMPK,p-AMPK,NF-κBp65 and NLRP3,ASC and caspase-1 in lung tissue.Results:1.HSYA could significantly inhibit the infiltration of inflammatory cells,goblet cell proliferation and mucus secretion in the pulmonary airways of asthmatic mice(P < 0.05).2.HSYA treatment group decreased the number of inflammatory cells in BALF of asthmatic mice in a dose-dependent manner,and HSYA could inhibit the number of total cells,lymphocytes,neutrophils,and eosinophils in BALF(P < 0.05).3.HSYA could increase the level of IFN-γ and decrease the levels of IL-4,IL-5,and IL-13 in lung tissue(P < 0.05).4.HSYA could activate AMPK phosphorylation levels and inhibit the protein levels of NF-κBp65,NLRP3,ASC and caspase-1 in lung tissue(P < 0.05).Conclusion:1.HSYA may alleviate airway inflammation in asthmatic mice by regulating Th1/Th2 immune imbalance;2.HSYA can reduce airway inflammation in asthmatic mice through the AMPK/NF-κB/NLRP3 signaling pathway.