Matrine On Prevented Hypertrophic Scar Of Rabbit Ears And On The Insulin Functions Which Promoted Proliferation And Collagen Synthesis Of Rat Cardiac Fibroblasts

Part 1: The experimental research of matrine on prevented hypertrophic scar of rabbit earsBackbround of experimentThe hypertrophic scar (HS) is one of the patterns by which the injuring skincorium repaired excessively. It forms hard tumeur that came from hyperplasied connective tissue (CT), and it most occurs in chest, back and four limbs of the body. The shape and function of organ may be changed by the HS. The patients may feel unbearable tickle and pain caused by cicatricial contration. It is the extracellular matrix (ECM) such as collagen, fibronectin and glucoprotein that composed the fundament of patholoty in the reparative process of skin. This EMC is synthesized excessively by fibroblasts. Whether generate and how extent of HS, it depends on the balance between of ecto-matrical synthesis and degradation. We can inhibit collagnic systhesis and cross-link, promot epithelization, collagnic degradation and regulated cytokine to therapy the HS. We commonly use the methods to therapy theHS such as antinotheaphy, crymotheraphy, exairesis and drug injection, but these methods have no ideal curative effect. We commonly use some medicines in the clinical such as corticoid, antihistamics, anti-free radical medicine, calciumion channel blocker, and Chinese gerbal medicine. It is the corticoid most affirmation in HS in the clinical. But so many complications are caused by corticoid in clinical, such as chromatosis, granula of drugs deposited, infection, necrosis, allergic response, menstrual disorder or amenorrhea in gynaec patient. These limited the application of corticoid. It has long time use Chinese herbal medicine to treat HS. The Chinese doctors consider it is the mechanism that the HS is caused by Ql-blood clogged, meridian clogged, or these factors complement each other. In our country each practitioner of Chinese medicine use different herbs to treat HS. And there are three methods to treat the HS in our country. One is endotherapy, the other is external treatment, and the last is the both. There has many ascendancy of external treatment, the first is the action is direct, the second is the operation is easy, the third is the curative effects is sure.It was reported that the synthesis of DNA and collagen in cultured scar-derived fibroblasts could be inhibited by tetrandrine in dose-dependent pattern.The Asiaticoside can significantly affect the ultrastracture of fibroblast and inhibit its collagen synthesis in a manner of dose-effect relationship.The fibroblast morphology can be altered by concentrationof Triperygium extract (LLZ) solution. Furthermore, the cell count and the cellular proliferating activity are all decreased obviously by the treatment of LLZ.The G2-M phase of fibroblasts is increased and the G2-M phase is prolonged by salvia miltiorrhiza when it's concentration is above 0.8mg/ml. The multiplication time of fibroblasts is prolonged by salvia miltiorrhiza. The mitosis of cells and the cellular cycle is inhibitory by salvia miltiorrhiza, and fibroblasts are still in G2-Mphase quiescently.The tetramethyl pyrazine has same effects on fibroblasts as salvia.Matrine can accelerate the apoptosis of hypertrophic scarring of human and model in rabbit ears. And matrine can decrease the proliferation, mitosis and the expression of proliferation cell nuclear antigen (PCNA) of fibroblasts. Matrine increases the percentage of S, G0-G1 phase and decrease the percentage of G2-M phase of fibroblasts. But the contents of DNA of fibroblasts is stabilized. Under the electron microscopy we can found that matrine can change the ultrastructural of fibroblasts: the rough endoplasmic reticulums are decrease, the ribosomes are degranulation and depolymerization, lysosomes are increase. The chromosomes condense and gather to the border of nuclear membranes. It is found in clinical the hypertrophic scarring change to the stable scarring by injected matrine .But the treatments of herbs on HS are experience. Furthemore there is a not effective Chinese formulated product on HS. There has no report that the external treatment is effective on HS. There has more effective on HS if the Chinese crud drugs is early used to. There still has no report that the Chinese crude drugs can prevent the development of HS.We will use the matrine on the surface of wound before HS formed. And wewill expect to prevent the HS generated.Objective:To investigate the effect of matrine on prevented hypertrophic scar of rabbit ears. Method:Established animal model of hypertrophic scar in 20 rabbit ears where 80 surfaces raw be made. The rabbits were divided into matrine group and normal saline group randomly, each group having 5 rabbits with 40 raw surfaces totally. From the second day of operation on the rabbit ears, matrine (lmg/ml) or normal saline was applied on rabbit ears. The volume of 0.5ml was given each time, threetimes a day. The hypertrophic scars were eccoped after three weeks, and made in HE stain, Masson stain, and pictric acid—sirus red stain respectively. Observation index: the heal time of normal heal scars, the numbers of hypertrophic scar, the collagenousdensity and I /III type collagenous density on area were observed. Result:Normal healed scars in matrine group were 12, and average healed time were10.6+1.7 days; 9 scars were normal heal in the saline group, and the average heal time were 10.1 + 1.1 days; there is no statistical significance (P>0.05). The numbers of hypertrophic scar in matrine group were 28 pieces, and 31 pieces in normal saline group; while there is no statistical significance between two groups. The average collagenous density on area of matrine group was 0.481+0.0481, normal saline group was 0.604+0.0403, there being statistical significance(P<0.001). The average I /HI type collagenous density on area of matrine group was 2.574+1.006, normal saline group was 2.286 + 0.4161, there being statistical significance(P<0.001)between two groups. Conclusion:The applied matrine can not prevent emergence of hypertrophic scars in rabbitears, but can lessen the degree of hyperplasia.Part 2: Effects of matrine on the Insulin functions which promoted proliferation and collagen synthesis of rat cardiac fibroUastsBackground:It was found that hyperinsulinemia mediated the reabsorption of water andsodium by kidney, which bring hypertension in directly or indirectly. The sympathetic system and renin-angiotensin system could be excited by hyperinsulinemia; and the level of endothelin (ET) also could be heighten by hyperinsulinemia.On the vitro experiment it was found that the cardiac fibroblasts could be proliferated and myocardial cell protein content (inducement myocardial cell portliness) could be increased by insulin, so the hypertension, cardiac pachynsis, myocardial fibrosis may be caused by hyperinsulinemia.Many organize fibrosis could be resisted by Chinese traditional medicine matrine. The function of angiotensin II which evoke the proliferation of cardiac fibroblastic could be resisted by matrine.It is not reported the function of insulin which could evoked cardiac fibroblastic proliferation could resisted by matrine. ObjectiveTo investigate the effects of matrine on the insulin functions which promotedproliferation and collagen synthesis of rat cardiac fibroblasts (CF). MethodsIsolated and cultured CFs of neonatal Sprague-Dawley (SD) rats were isolatedand cultured. Experiments were divided into five groups: ?control group: only with culture fluid; ?insulin group: with insulin (10~8mol ? L"1); ?low dosage Mat+Ins group: O.lmmol ? L'1 Mat+Ins; ?middle dosage Mat+Ins group: 0.2mmol ? L"1 Mat+Ins; ?high dosage Mat+Ins group: 0.4mmol 'L'1 Mat+Ins. CF proliferation wasmeasured by thiazolyl blue (MTT) assay. Collagen synthesis was measured byH-proline uptake test. Results?MTT colorimetry showed that the ,4490 values were 0.276+0.038 -. 0.140 +0.026, 0.089±0.005 respectively in the low, middle and high Mat+Ins groups, all significantly lower than that of the Ins group(0.869+0.108, all P < 0.05); and the middle, high Mat+Ins groups were significantly lower than that the control group(0.323+0.097, P < 0.05). ?The 3H-TdR uptake in the low, middle and high Mat+Ins groups was 964.76+56.7K 655.96+52.57and 376.14+56.33respectively , all significantly lower than that of the the Ins group (3882.95+661.08 , all P < 0.05); and the middle, high Mat+Ins groups were significantly lower than that the controlgroup(1326.32±278.03, P < 0.05). Conclusion:Mat inhibits effectively the function of Ins witch could promoted proliferationof cultured rat CFs and the collagen synthesis therein, and this contribution may enforce with the Mat concentration stepped up.