Effect Of Berberine Hydrochloride On Biobehavior Of Human Periodontal Ligament Cells And Expression Of TNF-α,IL-1β In Periodontal Tissues In Rats

Objective:1. To study the primary culture of human periodontal ligament fibroblasts (HPDLCs) by covering coverslip and to determine the influences of berberine hydrochloride on HPDLFs in primary culture.2. To determine the effects of berberine hydrochloride on the expressions of TNF-α,IL-1βin periodontal tissues in rat periodontitis model.Methods:1. Morphological analysis and immunocytochemical staining were used to identify cells origin and the efficiency of the primary culture by covering coverslip. A series of experiments were performed to study the effects of berberine hydrochloride on the biobehaviors of HPDLFs in vitro. Cell growth assay was used to evaluate the biological features of HPLF. Cell proliferation was determined using MTT assay, the total protein synthesis was detected by Coumassie Bright Blue method and the alkaline phosphatase activities were measured by spectrophotometric assay.2 . Based on the successful rat periodontitis model, the experimental rats were randomized into different groups followed by oral treatment with berberine hydrochloride for 1,2,3,4 weeks and then the rats were sacrificed and analyzed. Pathological assay and HE staining were used to detect the general conditions and pathological changes of rat periodontal tissues. And immunohistochemical staining was conducted to determine the expressions of TNF-α,IL-1βin rats periodontitis model periodontal tissues.Result:1.Covering coverslip increased the yield of primarily cultured HPDLCs significantly.2.The effects of berberine hydrochloride on the proliferation of primarily cultured HPDLFs: Compared to the control group, the proliferative abilities of PDLC were increased significantly when treated with 5,10,20,30mg/L of berberine hydrochloride for 24 hours, 5,10,20mg/L of berberine hydrochloride for 48 hours and 10,20mg/L of berberine hydrochloride for 72 hours (P<0.05).3.The effects of berberine hydrochloride on he the synthesis of total proteins: Compared to the control group, the total amount of proteins of PDLCs treated with 5,10,20mg/L of berberine hydrochloride was increased dramatically (P<0.05). 4.The effects of berberine hydrochloride on the ALP activity of HPDLCs: Compared to the control group, ALP activities of PDLCs treated with 10,20mg/L of berberine hydrochloride for 72 hours were enhanced significantly (P<0.05).5.The levels of TNF-α, IL-1βin the periodontitis tissues were significantly higher than that in the control group. Treatment with berberine hydrochloride decreased the levels of TNF-α,IL-1βin periodontitis tissues (P<0.05). Moreover, the general conditions and pathological changes in the control group and groups treated with berberine hydrochloride were much better than that in periodontitis groups.Conclusion:Covering coverslip increased the yield of primarily cultured HPDLCs significantly. Berberine hydrochloride at the dosages ranging from 10 to 20 mg/L promoted cells proliferation and the total amount of proteins and ALP activity of HPDLCs. Also it inhibited the expressions of TNF-α,IL-1βin periodontal tissues in rats periodontitis model and promoted the regeneration of the periodontal tissues. Taking together, our studies suggested that berberine hydrochloride may have potential clinical application.