Experimental Study On The Immunocompetence And Therapeutic Effect On Renal Fibrosis Of Amygdalin

PartⅠ:Amygdalin affects the human lymphocyte proliferation and the secretion of cytokine in vitro.Objective: 1.To observe the effect of amygdalin on human lymphocyte proliferation in vitro. 2.To research the influence of amygdalin on the secreting of IL-2,γ-IFN and TGF-β1 by human lymphocyte in vitro. Methods: The influence of amygdalin of different dose was detected by MTT assay on human lymphocyte proliferation in vitro. The level of IL-2,γ-IFN and TGF-β1 in supernatant of human lymphocyte culture was detected by ELISA. Results: 1.Amygdalin promoted the lymphocyte proliferation stimulated by PHA during the doses from 25μg·mL-1 to 800μg·mL-1. The promoting effect of amygdalin was the strongest at the concentration of 200μg·mL-1, and the value of stimulation index was 1.67. 2. Amygdalin promoted lymphocyte to secrete IL-2 andγ-IFN, but suppressed it to secrete TGF-β1, which was in a dose-dependent manner. Conclusion: Amygdalin could promote human peripheral lymphocyte proliferation stimulated by PHA and promote lymphocyte to secrete IL-2 andγ-IFN, but suppresses it to secrete TGF-β;by which it reinforces the immune function of recipients.PartⅡ:Therapeutic effect of amygdalin on renal fibrosisExperiment One: Inhibition of human renal fibroblasts proliferation by amygdalinObjective: The effect of amygdalin was invested on human renal fibroblasts proliferation. Methods: Human renal fibroblasts isolated from human kidney papilla were grown up in culture medium. They were identified by immunocytochemical staining. The inhibition of human renal fibroblasts proliferation by amygdalin was detected by MTT colorimetric assay. Results: After human renal fibroblasts were cultured for 24h, there was no difference between control group and the groups of amygdalin. After cultured for 48h or 72h, human renal fibroblasts proliferation was inhibited in a dose–dependent manner by amygdalin of doses from 25μg·mL-1 to 400μg·mL-1.The inhibiting effect of amygdalin is the strongest at dose of 100μg·mL-1, and the inhibition ration is 31.53% and 17.41%. Conclusion: Amygdalin could inhibit human renal fibroblasts proliferation in a dose-dependent manner.Experiment Two: Therapeutic effect of amygdalin on renal tubulo- interstitial fibrosis in ratObjective: To explore the therapeutic effect and mechanism of amygdalin on the renal tubulo-interstitial fibrosis induced by unilateral ureteral obstruction in rats. Methods: Renal tubulo-interstitial fibrosis was induced by unilateral ureteral obstruction in rats. Sixty Wistar rats were randomly divided into sham operation, unilateral ureteral obstruction and amygdalin treated groups, and they were sacrificed on day 7, 14 and 21 postoperatively. Pathological lesion of the rats was observed. Results: Renal fibrosis of all groups except sham operation became more and more serious along time, There was no difference on renal pathologic lesions between amygdalin-treated rats and those inflicted with unilateral ureteral obstruction on day 7 and 14. The renal pathologic lesions were significantly alleviated in amygdalin-treated rats as compared with those inflicted with unilateral ureteral obstruction on day 21. Conclusion: Amygdalin could obviously relieve renal fibrosis in rat.