| Objectives:1. To study the influence of person pure high triglyceride blood serum (HTG)on the prolifera -tion of rat aorta pectoralis blood vessel smooth muscle cells (VSMCs) , and discuss the biology changes in the cells under this pathological state.2. To discuss the protective function and the molecular mechanism of heme oxygenase-1 /carbon monoxide (HO-1/CO) system on the proliferation of VSMCs which stimulated by HTG.3. To observe the effect of scutellaria baicalensis stem-leaf total flavonoid (SSTF) on the proliferation of VSMCs which stimulated by HTG, and discuss its possible mechanism.Methods:Rat aorta pectoralis VSMCs were serial subcultived in vitro and induced to proliferate by HTG, to observe the effects of haemin(Hm, a substrate and inducer of HO-1) , zinc protopor- phyrin-â…¨(Znpp-â…¨, an inhibitor of HO-1)as well as SSTF on the proliferation of VSMCs, and discuss its possible mechanism.The experiment divided into the normal blood serum group (C group), the high triglyceride blood serum group (HTG group), the zinc original porphyrin group (HTG+Znp group), the concentration gradient heme group (HTG+Hm group), the pure heme group (C+H3 group) and the concentration gradient drug group (HTG+TF group). Used the tetrazolo colorimetric method (MTT) to examine the multiplication of VSMCs; applied the Flow Cytometry to demonstrate the cell mitotic cycle and multiplication index; emploied the transmission electron microscope technology to observe the changes in cell ultrastructure; handled the euzymelinked immunosorbent assay to detect the concent of carbon oxygen hemoglobin (COHb) in the raise supernate; the protein expression of HO-1 and the ERK/p-ERK were through Western blot.Results:1. Results from the MTT assays showed that compared with C group, the absorbance value (A value) increased remarkably by addition of HTG (P<0.01); Compared with the HTG group, the A value of HTG+Znp group remarkable increase (P<0.01), the A value of HTG+Hm group obviously reduced (P<0.01), and assumed the time and dosage dependence. But mon-addition of the greatest density Hm, compared with C group, A value of C+H3 group had no obvious change (P>0.05). SSTF can obviously suppress the cell multiplication by the HTG's induction (P<0.01), assumed the time ang dosage dependence.2. Results from the Flow Cytometry demonstrated that the progress of VSMCs mitotic cycle was stimulated obviously by addition of HTG. Compared with C group, the G0/G1 phase was obvious reduction, the S and G2/M phase obviously increased, the multiplication index also obviously elevated(P<0.01, P<0.05); more remarkable irritation of cell cycle progress by addition HTG and Znpp-â…¨(P<0.01). On the contrary, obvious suppressor of cell cycle progress by addition HTG and the concentration gradient of Hm, compared with the HTG group, G0/G1 phase increasesd, the S and G2/M phase reduced, the multiplication index also decreased (P<0.05, P<0.01), assumed the dosage dependence. By mon-addition of the greatest density Hm, the mitotic cycle change was similar to C group (P>0.05); the mitotic cycle progress of VSMCs were suppressed obviously by addition of SSTF (P<0.01), but the suppression of HTG+TF group was not as good as HTG+H3 group (P<0.05, P<0.01).3. Observes from the transmission electron microscope revealed that in normal blood serum group, the cell nucleus of VSMCs was big, assumed ellipse, sublobe or anomalous shape, the cell organs were rich, the intracytoplasm contained bulk of reticulate microtubules. In HTG group, cell nucleus karyotheca hollowed in different degree, the rough endoplasmic reticulum, ribosome and plastiosome increased obviously, some rough endoplasmic reticulum assumed cystic expansion, lots of sizes varied fat drops could saw in the intracytoplasm, pinocytosis bullules were under the cell membrane, but little myoneme could saw.4. Compared with the Control group, the COHb content was non-significance change when mono-addition of HTG (P>0.05) but increased obviously with mono-addition of Hm (P<0.01); compared with the Control group and HTG group, the COHb content increased obviously by addition of the concentration gradient Hm, assumed the time and dosage dependence (P<0.01), but the Znpp-â…¨group obviously reduced the COHb content (P<0.01). The SSTF group could enhance the production of COHb obviously (P<0.01), but the HTG+TF group is inferior to the HTG+H3 group on cell COHb output (P<0.01).5. Results from the Western blot displaied that the Control group, HTG group and HTG+TF group had few expression of HO-1 protein, comparison during two groups was not remarkable difference (P>0.05), the HTG+ZnP group complete suppression the expression of HO-1 protein. Compared with the Control group and HTG group, the expression of HO-1 protein were remarkable markup in Hm groups, assumed the dosage dependence (P<0.01). Moreover, compared with the Control group, the expression of ERK/p-ERK protein were obvious increased in the HTG group (P<0.01). Compared with the HTG group, the expression of ERK/p-ERK protein were more obvious increased in HTG+ZnP group (P<0.01). On the contrary, the expression of ERK/p-ERK protein were obvious decreased in the Hm groups, assumed the dosage dependence (P<0.01), and the same results occurred in HTG+TF group (P<0.01).Conclutions:1. The pure high triglyceride blood serum obviously promoted the multiplication of VSMCs, obviousiy irritated the progress of cell cycle, induced the cell microstructure to change.2. The HO-1/CO system play a protective function on the multiplication of VSMCs which stimulated by the pure high triglyceride blood serum, the activation of ERK signal passage may concern with this function.3. SSTF could depressed the multiplication of VSMCs which stimulated by the pure high triglyceride blood serum, and this inhibitory action was possible related to the activation of ERK signal passage.
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