Experimental Study Of Qiqirigan-8 On Atherosclerotic Inflammation By Regulating Macrophage Polarization

Objective:To investigate the effects of Mongolian medicine Qiqirigan-8 on macrophage polarization,and then to observe the effect of Qiqirigan-8 on atherosclerotic inflammation.Methods:72 Wistar male rats were randomly divided into 6 groups,each group has 12 rats,including a normal control group,an atherosclerosis model group,and the first group of Qiqirigan-8(Dosing when establishing animal model),and the second group of Qiqirigan-8(Dosing after establishment of animal model),the first group of simvastatin(Dosing when establishing animal model),the second group of simvastatin(Dosing after establishment of animal model),each group is given the corresponding feed and medicine,continuously rearing for 16 weeks.1.At 0,4,8,12,and 16 weeks,the body weight and general state of rats in each group were observed.2.To observe the effects of Qiqirigan-8 on blood lipid levels and aortic pathological changes in atherosclerotic rats.3.To observe the effects of Qiqirigan-8 on liver function(ALT,AST)in atherosclerotic rats.4.To observe the effects of Qiqirigan-8 on macrophage polarization in atherosclerotic rats:(1)Detection of macrophage phenotypic markers by ELISA,and M1 macrophage phenotypic markers are IL-6,TNF-a,i NOS;M2 macrophage phenotypic markers are Dectin-1,IL-10,Arg-1.(2)Immunohistochemistry method used CD68 to mark all macrophages,i NOS to mark M1 macrophages,and CD206 to mark M2 macrophages,and observed the distribution of macrophages according to the expression.(3)According to the RNA-Seq data,observe macrophage polarization related signaling pathways and verify the expression of related genes.Results:1.Results of body weight and general states of rats in each group:during the experiment,the normal control rats had smooth mental fur,normal activities,and good mental states;Model group rats have rough fur,less activities and poor mental states;The first group of Qiqirigan-8,the second group of Qiqirigan-8,the first group of simvastatin,the second group of simvastatin have smoother fur,activitiies and mental states were improved.No significant difference in body weight between rats in each group(P>0.05).2.Results of blood lipid levels and aortic pathological changes :On the 12 th weekend of the experiment,compared with the normal control group,the levels of serum TC,TG,LDL-C in the model group rats were significantly increased(P <0.05),and HDL-C levels were significantly decreased(P <0.05);The aortic pathological section showed that the intima of the model group was not smooth,and a large number of foam cells,lipid deposition,plaques protruded into the blood vessel cavity,and irregular proliferation of the median smooth muscle cells were seen in the model group,which proved that the model was established.On the 16 th weekend of the experiment,compared with the normal control group,the levels of serum TC,TG,LDL-C in the model group rats were significantly increased(P <0.05),and HDL-C levels were significantly decreased(P <0.05);Compared to the model group,serum TC,TG,LDL-Clevels were significantly decreased in the first group of Qiqirigan-8,the second group of Qiqirigan-8,the first group of simvastatin,and the second group of simvastatin(P < 0.05),HDL-C levels were significantly increased(P <0.05);Compared with the first group of simvastatin,the serum TC and LDL-C levels of the first group of Qiqirigan-8 were significantly decreased(P <0.05));Aortic pathological sections show,compared to the model group,the aortic pathological changes of the first group of Qiqirigan-8,the second group of Qiqirigan-8,the first group of simvastatin,and the second group of simvastatin improved to varying degrees.3.Results of serum liver function(ALT,AST): compared with the normal control group,the levels of serum ALT and AST in the model group were significantly increased(P <0.05);Compared to the model group,serum ALT and AST levels of rats in the first group of Qiqirigan-8 and second group of Qiqirigan-8 were significantly decreased(P <0.05);Compared with the first group of simvastatin and the second group of simvastatin,serum ALT and AST levels of rats in the first group of Qiqirigan-8 and second group of Qiqirigan-8 were significantly decreased(P <0.05)(1).4.Effect of Qiqirigan-8 on macrophage polarization in atherosclerotic rats ELISA test results:compared with the normal control group,the levels of IL-6,TNFa and i NOS in the model group rats were significantly increased(P <0.05),Dectin-1,IL-10,Arg-1 levels were not significantly different(P> 0.05);Compared to the model group,serum IL-6,TNF-a,and i NOS levels were significantly decreased in the first group of Qiqirigan-8,the second group of Qiqirigan-8,the first group of Simvastatin,and the second group of Simvastatin,Dectin-1,IL-10,Arg-1 levels were significantly increased(P <0.05).(2)Immunohistochemical test results : under the light microscope,compared with the normal group,a large number of brown deposits can be seen in the model group,the first group of Qiqirigan-8,the second group of Qiqirigan-8,the first group of Simvastatin,and the second group of Simvastatin,which represent CD68,i NOS,and CD206 are positive expression(P <0.05);Compared to the model group,the expression of CD68 in the first group of Qiqirigan-8,the second group of Qiqirigan-8,the first group of Simvastatin,and the second group of Simvastatin was significantly decreased(P <0.05),the expression of i NOS was significantly decreased(P <0.05),the expression of CD206 significantly was increased(P <0.05).(3)RNA-Seq results show that Mongolian medicine Qiqirigan-8 resists atherosclerosis through MAPK,IL-17,P13K/Akt and TNF signaling pathways,of which P13K/Akt signaling pathway is involved in process of the macrophage polarization.(4)Gene Irf1,Cxcl10 expression:in the arterial tissue,compared with the normal group,the expression levels of Irf1 and Cxcl10 genes in the model group were significantly increased(P <0.05);Compared with the model group,the expression of Irf1 and Cxcl10 in the first group of Qiqirigan-8,the second group of Qiqirigan-8 were significantly decreased(P <0.05).Conclusion:1.Mongolian medicine Qiqirigan-8 can regulate blood lipid levels.2.Mongolian medicine Qiqirigan-8 has the effect of reducing ALT and AST levels and protecting the liver.3.Mongolian medicine Qiqirigan-8 can regulate macrophage polarization,from the pro-inflammatory M1 to the anti-inflammatory M2.4.Mongolian medicine Qiqirigan-8 may regulate macrophage polarization through Irf-1 and Cxcl10 genes.5.Mongolian medicine Qiqirigan-8 has the function of regulating metabolism,promoting blood circulation and removing blood stasis to resist inflammation caused by indigestion of essence and blood "yellow water" stasis in the veins.