| Objective: 1.We systematically summarized the theory of “microcosmic zhengji”.Furthermore,we summarized the experiences of the diagnosis and treatment of TCM on cardiovascular disease under the guidance of this theory.2.We researched the early therapeutic effects of oridonin in atherosclerosis(AS)rats.And the intrinsic mechanism of oridonin on AS was that it induced autophagy through PI3K/Akt/mTOR signaling pathway.And which was in order to clarify and enrich the scientific connotation of the theory of“microcosmic zhengji”.Method:1.We first summarized the origin of the theory of “microcosmic zhengji” from the literature researches and the relationships between AS and tumor.Immediately,we systematically summarized the connotation and significance of “microcosmic zhengji”theory.Furthermore,we summarized the clinical experiences of TCM in the diagnosis and treatment of cardiovascular diseases using the method of typical medical case appreciation.2.(1)In vivo experiments: Forty male SD rats were randomly divided into normal control group(C group),model group(M group),oridonin prevention group(O group)and simvastatin Prevention group(S group),and which each group had 10 rats.Experimental rats were used to establish models of early atherosclerotic plaque by intraperitoneal injection of vitamin D3 and high fat diet.The research methods were early intervened withdrugs: Oridonin group and simvastatin group were both administered by 5mg /(kg·d),while the C group and M group rats were given equal volume of saline.After 12 weeks,the experiment was ended.The degree of atherosclerosis in each group were detected by HE staining and oil red O staining.And the changes of body weight,serum lipids,AI index and serum TNF-? levels were also detected.Furthermore,the expression of autophagy protein LC3 was detected by tissue immunohistochemistry(IHC)and immunofluorescence(IF).(2)In vitro experimental studies: We used Ang-? to stimulate the proliferation of rat arterial smooth muscle cells(RASMCs).Firstly,the optimal concentration of Ang-? and the best intervening concentration of oridonin were determined by CCK-8 assay.The main groups of experiments were divided into Control group,model group(Ang-? group),oridonin group(ORI group),simvastatin treatment group(Ang-?+ SIM group)and oridonin treatment group(Ang-?+ ORI group).Then,cells migration and invasion were evaluated by wound healing assay and transwell chamber assay.The expressions of autophagy-related proteins LC3?/? and Beclin-1 were detected by Western blot and IF assay.Furthermore,we researched the internal pathway through which the oridonin inhibited the excessive proliferation and invasion of RASMCs.Firstly,RASMCs were treated with 3-MA which was an inhibitor of autophagy.The expressions of LC3 in each group were detected by Western blot and IF.Secondly,western blot was used to detect the protein expression of PI3 K / Akt / mTOR which was an autophagy related pathway.Result:1.It was stated that there were many similar etiological and pathological changes in the predisposing factors and pathogenesis between AS and tumors through literature research.There were many cross-similarities in the expression of physico-chemical factors,such as inflammatory factors,growth factors,chemokines,adhesion factors,and proto-oncogenes.In the clinical practice of TCM on the coronary heart disease,tonifying qi and Huaji and promoting blood circulation and detoxication were as the main treatment principle based on the theory of “microcosmic zhengji”.Thus,the oridonin which had significant clinical efficacy.2.(1)The results of experiments in vivo: Compared with C group,the levels of TC,TG,LDL-C,TNF-? and AI index were significantly increased in M group.In the M group,the arterial lumen of the rats presented with different degrees of stenosis,roughness,rupture of the intima-media elastic sheet,elastic fiber degeneration,crack,endothelial cell damage and thickening,a large number of foam cell deposition,lipid droplets deposition and oil red O staining significantly.But compared with M group,the levels of blood fat,TNF-? and AI index in O group and S group were significantly decreased.And compared with group M,the pathological changes of arterial AS such as lipid deposition and endothelial cell injury in experimental group O and group S were light,and there were almost no early lesions of AS.Furthermore,Compared with group C and group M,the expression of LC3 in the aorta tissue of O group was increased which the difference was statistically significant.It was suggested that oridonin inhibited the early stage of AS by enhancing autophagy.(2)The results of experiments in vitro: When the concentration of Ang-? was 10-5mol / L,the proliferation rate of RASMCs was the highest.While Oridonin was 10 ug/ml,the inhibition of RASMCs was the maximum.Compared with Control group,the proliferation and migration of RASMCs in Ang-II group were obvious.Compared with Ang-? group,there was no obvious proliferation and migration pathways in ORI group,Ang-?+ ORI group and Ang-? + SIM group(P<0.01).Compared with Control and Ang-? group,the expressions of LC3?/? and Beclin-1 in Ang-?+ ORI group were higher.But the activity of oridonin on cell autophagy was significantly decreased after 3-MA intervention.It showed that the oridonin against AS was related to the induction of autophagy activity.Compared with the control group,the expressions of LC3?/? in the ORI group and the Ang-? + ORI group were significantly higher,which indicating that the autophagic activity of the cells was stronger.However,the expressions of P-PI3 K,P-Akt,P-mTOR and P-P70S6 K decreased with statistical significance.It is suggested that oridonin could inhibit the over-proliferation of RASMCs by increasing the autophagy activity through inhibiting the PI3 K / Akt / mTOR pathway.Conclusion:1.The theory of “microcosmic zhengji” proposed by Professor Song,whichatherosclerotic disease was associated with the disease of Zhengji for the first time.And it had enriched the guiding theories of traditional Chinese medicine in diagnosis and treatment of atherosclerotic diseases.In the diagnosis and treatment of atherosclerosis related diseases in Chinese medicine,the skillful use of anti-mass Chinese medicine can greatly improve the clinical effect.2.Under the guidance of “microcosmic zhengji” theory,oridonin could effectively reduce the levels of serum lipids,AI index and the inflammatory activity factor TNF-? in the rats fed with high fat diet,and prevent the development of atherosclerosis disease.3.Based on the theory of “microcosmic zhengji”,oridonin promoted autophagy activity by inducing the expression of autophagy-related proteins LC3? / I and Beclin-1to inhibit the RASMCs from excessive proliferation and metastasis.4.Oridonin inhibited the PI3 K / Akt / mTOR signaling pathway to promote autophagy activity of RASMCs,and inhibited the excessive proliferation and metastasis of RASMCs in order to prevent the development of AS. |
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