Mechanism Of Atherosclerosis Caused By Inside Invading Of Outside Evil And Interventing Effect Of Heat-Clearing And Detoxicating Remedy

1. ObjectiveThe purpose of this study was to establish animal model of Atherosclerosis (AS) of inside invading of outside evil by infecting CPn and fedding high cholesterol diet to mice, in combination with clinic study , to approach the mechanism of AS caused by inside invading of outside evil on lipid disorder, oxidative stress, fibrinolytic system . Attempts were made to observe the effect of heat-clearing and detoxicating remedy treatment to these index at the early stage of AS . To provide new method and idea to the preventing of AS.2. Methods(1)Animal experiment. We used C57BL/6J mice to establish animal model ofAtherosclerosis (AS) of inside invading of outside evil by infecting CPn (mice were inoculated intranasally with C. pneumoniae three times, every 2 weeks over a 6-week period) and fedding high cholesterol diet. To get mice with Hyperhomocysteine by adding methionine into the diet at the 8th week. We observed the effect of the CPn, high cholesterol diet and Hyperhomocysteine to AS. Berberine, azithromycin and atorvastain were drenched to observe their effect. Ber1 group were be given berberine at the beginning of CPn infecting .while ber2 group , azithromycin group and atorvastain group were be given the drug after the 3rd CPn infecting. At the 16th week , all mice were sacrificed for serum collection. The serum was used to test IL-1β, Hs—CRP and OX-LDL by ELISA method. TC(total cholesterol), TG(triglyceride), HDL-C(high-density lipoprotein) and LDL-C(low-density lipoprotein) were be also tested. Isolating ascending aorta and aortic arch to analyze the expression of HSP-70(Heat-Shock Protein-70) and count the monocytes infilitrating into the intima of the aortas. The aortas were be also observed with HE dye in normal microscope and electron microscope. (2)Clinic study. 50 patients were be selected and randomly divided into two groups, 33 in treated group and 17 in control group. According to the disease period , they were divided into with acute and chronic disease . Treated group were be givenSanHuang pill for 2 weeks addition to the base treatment with western medicine. At the beginning and end of the treatment ,their serum were be taken to test CRP(C-reactive protein), Hs~CRP(High sensitivity C-reactive protein), Hey (homocysteinemia), WBC, FIB(fibrinogen), D-D(D-dimmer), TC(total cholesterol), TG(triglyceride), HDL-C(high-density lipoprotein) and LDL-C (low-density lipoprotein). 3.Result (1)Animal experiment.The effect of Berl group on AI (Atherosclerotic Index) was correspond to atorvastain group, had an advantage over Ber2 and azithromycin group. The effect of Berl group on TG and HDL was correspond to azithromycin group. The effect of Berl group on TG had an advantage over other groups. In establishing AS model groups, high cholesterol diet and CPn could accelerate lipid disorder, in high cholesterol + CPn group, lipid disorder was the most. Lipid disorder in groups with Hyperhomocysteine (by adding methionine into the diet) conspicuously differed from groups without methionine diet.The effect on OX-LDL . Berl group was correspond to atorvastain group and azithromycin group, had an advantage over Ber2 group. Berberine had no effect on OX-LDL in high cholesterol group. The level of OX-LDL in cholesterol+ CPn group conspicuously differed from that in normal control group , normal diet + CPn group and high cholesterol + CPn group. The level of OX-LDL in normal diet + CPn group conspicuously differed from that in normal diet + CPn +hcy group.The effect on IL-1{3. Berl group was correspond to atorvastain group and azithromycin group, had an advantage over Ber2 group and Ber3 group. The level of IL-ip was conspicuously higher than that in other groups. There was no difference between groups with and without methionine diet.The effect on Hs—CRP. Berl group was correspond to azithromycin group, had an advantage over Ber2 group , Ber3 group and atorvastain group. In establishing AS model groups, with either high or normal diet, CPn could promot the expression level of Hs—CRP, which conspicuously differed from groups without CPn infection.Pathologic and electron microscope examination. The expression of HSP-70 in Berl group , azithromycin group and atorvastain group had no different, but had an advantage over Ber2 group , Ber3 group and high cholesterol + CPn group. Theexpression of HSP-70 in either methionine diet or not group were the same. The effect on the monocytes infilitrating into the intima of the aortas. There was conspicuously different among Berl group , Ber2 group , Ber3 group azithromycin group(correspond to atorvastain group )and high cholesterol + CPn group. The effect of Berl group had an advantage over other groups. Observing with normal microscope (by HE dye) and electron microscope, it was found that endotheliocyte process , tunica media destroyed, monocytes infilitrating and foam cell forming at various degrees in all groups. Observing with electron microscope, it was also found that chondriosome and lysosome was destroyed. Berl group , atorvastain group and azithromycin group had an advantage over other groups. (2) Clinic studyThe effect on lipid. Treated group had an advantage over control group onlowering TG, (TC—HDL) / HDL and TC / HDL. In each group, the level of lipid beforetreatment differed from that after treatment. In acute subgroup, the value of(TC —HDL) /HDL> TC / HDL> LDL/HDL were different between Treated group andcontrol group.The effect on Fib(fibrinogen) and D-D(D- dimmer). There were no difference between the two groups. In each group, the level of Fib and D-D before treatment differed from that after treatment. The same result occurred in acute subgroup.The effect on CRP (C-reactive protein) and Hs~CRP(High sensitivity C-reactive protein). There were no difference of CRP between the two groups before and after treatment. The level of Hs~CRP was different between treated group and control group , and Hs-CRP before treatment differed from that after treatment. The same result occurred in acute subgroup.The effect on Hcy(homocysteinemia). There were no difference between the two groups before treatment. After treatment, either in all the patients or in acute subgroup, the treated group had an advantage over control group.The effect on WBC. There were no difference between the two groups. 4. Cone I us i ons (1)Animal experimentHigh cholesterol diet and CPn could accelerate lipid and AI (Atherosclerotic Index) disorder in mice, these observations suggest that CPn infection can accelerate AS by accelerating lipid disorder. The effect on lipid and AI of earlytreatment with Berberine is correspond to atorvastain, has an advantage over late treatment with Berberine and azithromycin. Hyperhomocysteine can accelerate AS by accelerating lipid and AI disorder.The expression of OX—LDL was increased by high cholesterol diet and CPn, these observations suggest that CPn infection can accelerate AS by oxidative stress. The effect on lipid and AI of early treatment with Berberine is correspond to atorvastain and azithromycin, has an advantage over late treatment with Berberine. The expression of OX—LDL was increased in normal diet + CPn +hcy group, the observation suggest that CPn infection can increase the reaction of oxidative stress in cooperation with hyperhomocysteine. But in high cholesterol group, hyperhomocysteine hasn't the cooperation effect.The expression of IL-ip was increased by high cholesterol diet and CPn, these observations suggest that CPn infection can accelerate AS by increasing the expression of inflammatory factor. The effect on IL-ipof early treatment with Berberine is correspond to atorvastain and azithromycin, has an advantage over late treatment with Berberine. In establishing AS model, hyperhomocysteine has no effect on IL-ip.The expression of Hs-CRP was increased by CPn either in high cholesterol diet or normal group. The effect on Hs-CRP of early treatment with Berberine is correspond to azithromycin, has an advantage over late treatment with Berberine and atorvastain. CPn infection can increase the expression of Hs-CRP in cooperation with high cholesterol and hyperhomocysteine.CPn infection and hey has no effect on HSP-70(Heat-Shock Protein-70) by half-estimation. The effect on HSP-70 of early treatment with Berberine is correspond to atorvastain and azithromycin, has an advantage over late treatment with Berberine. High cholesterol diet cooperated with CPn infection, and then with hey, can increasing the monocytes inf ilitrating into the intima of the aortas. On which early treatment with Berberine has an advantage over other groups. (2) Clinic studyThe treatment of heat-clearing and detoxicating remedy has effect on lipid disorder. Especially in acute subgroup, this type of treatment has better effect on TG and HDL than control.The treatment of heat-clearing and detoxicating remedy has effect on AIVII(Atherosclerotic Index), especially in acute subgroup.The treatment of heat-clearing and detoxicating remedy has effect on FIB and D-D, inhibit the activation of fibrinolytic system, but the effect has no difference with control.Hs—CRP is better than CRP in reflection of inflammation . The treatment of heat-clearing and detoxicating remedy has good effect on hs-CRP, especially in acute subgroup.The treatment of heat-clearing and detoxicating remedy has good effect on HCY, which suggest that the treatment can relieve the inflammatory reaction in AS and decelerate development of AS.