The Study On The Effect Of Shaoyangshenggu Formula On Proliferation Of Rat Chondrocytes And Its Collagen Type â…¡ Expression

objective: By comparing the observed ShaoYangShengGuFang(SYSGF) and glucosamine hydrochloride capsule on rat articular cartilage cellproliferation in vitro, the expression of the type II collagen, explore theprotection mechanism of SYSGF on impaired articular chondrocytes. Methods:1.Establish rat articular chondrocytes in vitro and identification:The primary chondrocytes were got from one-week old SD articularcartilagewith collagenase and trypsin digestion, observed the morphology under inverted microscope,identified the biological characteristics of chondrocytesby toluidine blue staining.2. MTT assay was performed to determine the effect of SYSGF and glucosamine hydrochloride capsules containing medicine serum on proliferation of normal chondrocytes．3. Immunohistochemistry was performed to observe the effect of SYSGF and glucosaminehydrochloride capsules containing medicine serum on expression type IIcollagen of normal chondrocytes. Results:1.Establish rat articular chondrocytes in vitro culture system with collagenase and trypsin digestion,andobserve the biological characteristics of chondroeytes with inverted microscope, and identified by toluidine blue staining, And confirm the separation of cultured cells in cartilage cells of biological characteristics, morphology, histochemical staining showed cartilage cell culture can maintain the stability of phenotypic within three generations.2.Observation of the ca rtilage cell proliferation by MTT assay: The SYSGF were5%,10%,5%serum containing drug concentration can promote normal cartilage cell proliferation in vitro, but the proliferation effect increases with increasingconcentration,10%drug-containing serum concentration promotes normal articular cartilage cell proliferation is most obvious.And the48h time,promoting proliferation effect best.10%SYSGF10%medicated serumgroup and blank serum group compared with significant difference. Andthe48h time, for normal cultured chondrocytes, the10%concentrationof SYSGF was compared with the blank serum group, there was remarkable statistical difference(P<0．05).and when campareed control group, There was significant differences (P<0．05), The proliferation effect was less than the control group. at the24h time, all groups proliferation effect were weak.3．The result of imunohis tochemistry show as follows：All groups’ specimens expressed The type II collagen, and the expressionsite was only in the cytoplasm，with red-brown particles. the expressionof type II collagen blank serum group was lower than the SYSGF group,.there was remarkable statistical difference(P<0．05),and compared with controlled group, the expression of type II collagen was lower. Conclusion：SYSGF to normal articular cartilage cells has good role in promotingproliferation, can promote the synthesis of normal articular cartilage cells secrete type II collagen,and Prompt SYSGF has protective effect on articular cartilage. This is the one of the mechanism of prevention and tr eatment of osteoarthritis.