Clinical And Experimental Study Of Dihuai Xiaoyin Granules In The Treatment Of Blood-heat Syndrome Of Psoriasis

BackgroundPsoriasis,one common chronic skin disease,which seriously impacts the physical and mental health of patients.Traditional Chinese medicine(TCM),which has a long history,has great understanding of the disease,and in the evolution of history,traditional Chinese hemo-modulating therapy rise in response to the proper time and conditions,which has got acceptance by increasing doctors.Blood heat syndrome is the most common syndrome type in hemo-modulating therapy.Dihuai Xiaoyin prescription has been used in clinic for many years and good curative effect has been achieved,besides,it would be more convenient if we make the prescription into granules.Recently,there are numerous discussion about the mechanism of traditional Chinese medicine,and the pathway of phosphatidylinositol 3-hydroxykinase/protein kinase B(PI3K/Akt)signal transduction is the current research hotspot,which do not only plays an essential role in the pathogenesis of tumor,but also play a significant role in the pathogenesis of psoriasis.Previous studies have confirmed that Dihuai Xiaoyin granules might play a role through this pathway.The purpose of this study was to verify the clinical efficacy of Dihuai Xiaoyin granules in the treatment of psoriasis with blood heat syndrome,and to explore the mechanism at the animal and cellular level based on the PI3K/Akt pathway.MethodClinical study:We adopted the randomized controlled observation that 72 patients who suffer from psoriasis with blood heat syndrome were randomly divided into observation group and control group.General information,skin lesion PASI score and TCM syndrome score were recorded.The patients in the observation group were given Dihuai Xiaoyin granules orally,and the patients in the control group were given psoriasis xataret tablets orally,besides,both groups would be treated with topical medical vaseline.The observation period was 56 days.PASI score and TCM syndrome score were recorded before and after treatment.To observe the effects of Dihuai Xiaoyin granule and psoriasis xatahot tablets on psoriasis with blood heat syndrome,which means that,the change of PASI score,TCM syndrome score and efficacy,besides,calculating their respective clinical effective rates.Animal experiment research:To build psoriasis BALB/c mouse model by externally using imiquimod,after which,we treated the mouse model with Dihuai Xiaoyin granular((high,medium and low dose group)by intragastric administration.And then,the enzyme linked immunosorbent assay(ELISA)would be used to detect the content of peripheral blood cytokines,which contain IL-2,IL-6,IL-8,IFN-? and TNF-?,besides,the method of Western blotting and RT-PCR would be used to detected the expression level of PI3K,Akt protein and mRNA in the lesion tissue.All we talked above were to discuss the influence of Dihuai Xiaoyin granule to the psoriatic mouse,the observation targets mainly contain the lesion tissue,cytokines,proteins related to pathways and gene,etc.Cell experimental study:Human adult low calcium temperature(HaCaT)was stimulated by epidermal growth factor(EGF)for 24 hours,and LY294002,specific blocker of PI3K/Akt signaling pathway,was added to intervene for 1 hour to prepare the cell model required for the experiment.And then,the HaCaT cell model,which had been stimulated by EGF and added with inhibitor,would be intervened by Dihuai Xiaoyin particles(high,medium and low dose group)respectively.After which,we observed the proliferation,apoptosis,cell cycle of the HaCaT cell model and the influence of the content of IL-2,IL-6,IL-8,IFN-?,TNF-?,besides,Western blotting and RT-PCR would be used to detect the expression level of PI3K,p-Akt,Akt protein and PI3K,Akt mRNA.ResultClinical study:After 8 weeks of treatment,a total of 66 patients were observed at last,the effective rate of Dihuai Xiaoyin granule was87.88%,higher than66.67%of the control group.The PASI score after treatment was significantly lower than that of the control group,and the difference was statistically significant(P<0.01).The improvement of TCM syndrome was better than that in the control group,and the difference was statistically significant(P<0.01).Animal experiment research:1.Dihuai Xiaoyin granules could ameliorate the skin lesions of mouse with psoriasis induced by imiquimod.The main effect on pathology is to improve epidermal hyperplasia and dermal vascular hyperplasia in psoriasis and reduce inflammatory reaction;2.Dihuaixiao silver granule can inhibit the secretion of i1-2,il-6,il-8,IFN-,TNF-in serum of mice;3.We detected the PI3K mRNA and protein in mouse tissues,which drew a conclusion that the high-dose group was the most effective one.It could down-regulate the expression of mRNA gene and affect the production of protein,and this function was realized through the PI3K/Akt pathway,besides,it had no effect on Akt mRNA and Akt protein.Cell experimental study:1.Dihuai xiaoyin granule can inhibit the proliferation of HaCaT cells,and its effect could be weakened when LY294002 pathway was inhibited,and its inhibitory effect on cell proliferation is realized through the PI3K/Akt pathway;2.Dihuai xiaoyin granules can make cell division stagnating in G0-G1 or S period,which prevent cell proliferation,and can also increase the rate of late stage cell apoptosis.These effects were significantly inhibited after the addition of LY294002 the pathway inhibition;3.We detected the expression level of PI3K,p-Akt,Akt protein and PI3K,Akt mRNA through the method of Western blotting and RT-PCR,which drew a conclusion that Dihuai Xiaoyin granules could reduce the level of PI3K protein by down-regulating the level of PI3K mRNA,and these effects would disappear after the addition of pathway inhibitors,however,the level of Akt protein and Akt mRNA were stable.All we talked above demonstrate that Dihuai Xiaoyin granules could exert its effect by down-regulating the level of PI3K mRNA and thereby affecting the level of PI3K protein;4.Dihuai Xiaoyin granules could inhibit the secretion of IL-2,IL-6,IL-8,IFN-? and TNF-?,in which,the PI3K/Akt pathway may play an essential role,and this effect was obviously reflected in the secretion of IL-2,IL-8,IFN-? and TNF-?,while it may play a role in inhibiting the secretion of il-6through a variety of ways;5.Dihuai Xiaoyin granules act through the PI3K/Akt pathway,and have an effect on the PI3K gene and protein,through which,it affects the secretion level of a variety of cytokines such asIL-2,IL-6,IL-8,IFN-?,TNF-? and play a role in the treatment of psoriasis.ConclusionThe efficacy of Dihuai Xiaoyin granule in the therapy of psoriasis with blood-heat syndrome is certain with no adverse reactions occurred,which could ameliorate the severity of skin lesions of patients and reduce the TCM syndrome score;2.Dihuai Xiaoyin granule could ameliorate the skin lesions of mouse with psoriasis that induced by imiquimod,and the mechanism is achieved by down-regulating PI3K mRNA synthesis to inhibit protein production and cytokine production.3.Dihuai Xiaoyin granule could inhibit the proliferation of HaCaT cells through the PI3K/Akt pathway,it could also down-regulate the expression of PI3K mRNA,inhibit the synthesis of PI3K and pAkt proteins,and inhibit the secretion of a variety of cytokines.To sum up,Dihuai Xiaoyin granule has a definite clinical effect in the treatment of psoriasis with blood heat syndrome,and its specific mechanism may be to promote apoptosis and inhibit proliferation by down-regulating PI3KmRNA and protein synthesis and inhibiting the secretion of a variety of cytokines.