Jiawei Xiaochaihu Granule Of The Idiopathic Thrombocytopenic Purpura Clinical And Experimmental Research

Purpose:Primary idiopathic thrombocytopenic purpura(ITP) is an autoimmune disease, currentresearch shows endocrine and immune abnormalities constitute an important part of thepathogenesis. There is no effective means of the radical at present in modern medicine, andso there is a must to find effective methods to against ITP in TCM field. Primary ITP can beClassified as "Blood disease" in TCM field, we believe that mechanism of this disease isdisharmony of liver-qi, then it brings imbalance of qi and blood, discordance of liver andspleen, blockage of blood stasis. The modern TCM research shows that the function of theliver to smooth and regulate the flow of qi is closely related to the endocrine-immunenetwork, according to the pathogenesis and the pathological features of ITP, "from livergovemance" principles and "harmonize the liver-qi and get rid of the blood stasis"treatment is established. By using Xiaochaihu Tang which is from "Shanghanzabinglun"and at the same time adding blood stasis removing herb in it, finally it becomes JiaweiXiaochaihu Granule.The purpose of this research is via theoretical research, clinicalresearch and animal experiments, to discuss the effect of Jiawei Xiaochaihu Granule to ITPthrough the studying of endocrinology-immunology.Method:1. By using 60 ITP patients, randomly divided them into two groups, one was theobserver group and the other is the control group, each group contained 30 patients. For theobserver group was treated with prednisone together with Jiawei Xiaochaihu Granule, andcontrol group with Prednisone only. By observing the recent effect, long-term effect sixmonths after treatment, average onset time, average time of reducing the use ofPrednisone, rate of improvement in clinical symptoms and side effects of Prednisone andother related indexes via the evaluation of clinical efficacy, clinical symptom scores andside effects of using Prednisone. 2. Experimental study was divided into two parts, one was the clinical efficacy andthe other was the mechanism. ITP mice were selected to observe the effect of JiaweiXiaochaihu Granule to the platelet count, megakaryocyte, subgroups of T-cell,platelet-associated antibodies, glucocorticoid receptor, corticosterone and other indexes ofITP mice.(1) Production of ITP mice: Inject guinea pig anti-mouse platelet serum (APS), toestablish mice model of autoimmune thrombocytopenic purpura. Injecting diluted (1:4)antisera to mice by celio-injection in accordance with 100ul/10 grams in weight. At 6th hourafter injection, platelet declined, and reached its minimum at 8th hour. To maintain plateletsustained at a low level, inject anti-platelet serum in 1st,3rd,5th,7th,9th, 11th,13th,15th,17th,19thdays, and treat the mice according to the group which they belong to at the 7th day aftermodeling.(2) Acute toxicity experiment: To treat the mice with Jiawei Xiaochaihu Granule(98g/kg) that was in the largest concentration and volume through gavage in once, none ofthe mice died or showed obvious abnormalities. It suggests that the toxicity of JiaweiXiaochaihu Granule is relatively low, the lowest lethal dose of half of the mice couldn’t bedetected, and so it was changed into detecting the multiple dose administration of thelargest volume in one day.(3) Clinical efficacy of the treatment of ITP mice: Determine the peripheral bloodplatelet count by using the blood count meter before modeling and the treatment, at the4th,7th and 14th day after treatment. Observe the megakaryocyte cell count andclassification of ITP mice under the microscope and also test the time of bleeding bycutting tail of ITP mice to study the clinical effect of Jiawei Xiaochaihu Granule to ITPbefore and after the intervention of Prednisone.(4) Impact on endocrine-immune function of ITP mice: Use ELISA to detect theperipheral blood platelet-associated antibodies, flow cytometry to detect peripheral blood Tlymphocyte subsets, RIA to detect peripheral blood cytokines, radioligand binding analysisto detect and find the glucocorticoid receptor loci of spleen cell of ITP mice, RT-PCR todetect the expression of glucocorticoid receptor mRNA to clarify the significance ofabnormalities of immune-endocrine function to ITP and the clinical effect after JiaweiXiaochaihu Granule intervention.(5) The increase in clinical efficacy and reduction of toxicity after hormoneintervention:Determine the level of glucocorticoid receptor loci and glucocorticoid receptor mRNA, plasma ACTH、CORT by using RIA, clarify the effect of glucocorticoid toendocrine system of ITP and the regulatory effect of Jiawei Xiaochaihu Granule.Results:1. Clinical research: It was found that efficacy of the observer group better than thecontrol group (P＜0.05); the average onset time of treatment of the observer group was9.7±3.6 days, control group was11.5±3.7 days (P＜0.05); the initiation of reduction ofhormone of the observer group was 26.4±3.6 days, control group was 28±3 days (P＞0.05);the platelet count still more than 50×10~9/L six months after the treatment of the observergroup was 56.67%, control group was 26.67% (P＜0.05); rate of coming up of side effects ofthe observer group was lower than the control group (P＜0.05). It suggested that JiaweiXiaochaihu Granule can improve clinical efficacy, shorten onset time and reduce theincidence of side effects of hormone.2. Experimental research:(1) Acute toxicity experiment: The MTD (Maximum Tolerance Dose) of JiaweiXiaochaihu Granule of the mice was at least 194 times of clinical normal dosage. It suggeststhat Jiawei Xiaochaihu Granule doesn’t show apparent toxic reaction, and so it is safe andreliable for us to use.(2) Clinical efficacy of the treatment of ITP mice: The platelet count of ITP micecontinued to drop, bleeding time prolonged, the count of megakaryocyte increased butdecreased in the count of megakaryocyte which can generate platelet. There was a significantdifference when compared with normal group (P＜0.05). Jiawei Xiaochaihu Granule couldraise the blood platelet count, shorten the bleeding time, decrease the amount ofmegakaryocyte and increase the amount of megakaryocyte which can generate platelet. Therewas a significant difference when compared with normal group (P＜0.05) and better than themice treated with Weixuening Chongji (P＜0.05). It showed that the clinical efficacy of thegroup treated with hormone together with Jiawei Xiaochaihu Granule was better than thegroup treated with hormone only (P＜0.05).(3) Impact on endocrine-immune function of ITP mice: The level of platelet-associatedantibodies, CD8, IL-10 of ITP mice were increased (P＜0.05) but the level of CD3, CD4,CD4/CD8, IL-2, glucocorticoid receptor of spleen cell and its mRNA of ITP mice weredecreased when compared with normal group (P＜0.05). When consider mice which weretreated with Jiawei Xiaochaihu Granule, the level of plasma CD3, CD4, CD4/CD8, IL-2,glucocorticoid receptor of spleen cell and its mRNA increased but the level of platelet-associated antibodies, plasma CD8, IL-10 decreased when compared with the controlgroup (P＜0.05). It showed that Jiawei Xiaochaihu Granule can regulate the abnormalities ofendocrine-immune function of ITP mice.(4) Impact on the function of endocrine system after hormone intervention: The adrenalindex, level of plasma ACTH, CORT of the group treated with hormone only decreasedtogether with the decrease of the level of glucocorticoid receptor of spleen cell and its mRNA.There was a significant difference when compared with the ITP group (P＜0.05). When itcomes to the group which was treated with hormone together with Jiawei XiaochaihuGranule, those indexes mentioned before increased and had a significant difference whencompared with the group treated with hormone only (P＜0.05).It suggested that JiaweiXiaochaihu Granule can regulate the decrease in the level of glucocorticoid receptor of spleencell and its mRNA and also dysfunction of hypothalamic-pituitary-adrenal axis due tohormone intervention.Conclusion:1. Clinical test: Treating ITP with Jiawei Xiaochaihu Granule together with Prednisonecan increase the clinical efficacy, prolong the valid time of treatment and reduce the sideeffects of hormone.2. Experimental research: Not only the Jiawei Xiaochaihu Granule can significantlyincrease the platelet count of ITP mice, promote the maturation of megkaryocyte in bonemarrow, inhibit platelet-associated antibodies, regulate T-lymphocyte subsets and cytokineswhich are produced by T-lymphocyte, raise the level of glucocorticoid receptor loci andglucocorticoid receptor mRNA, protect the normal function of thehypothalamic-pituitary-adrenal axis, but also doesn’t show apparent toxic reaction.3. Both of the clinical and experimental research showed that Jiawei XiaochaihuGranule is effective to Primary ITP. Besides, it can also increase the efficacy and reduce thetoxicity or side effect of hormone and maybe due to the regulatory effect toendocrine-immune system of Jiawei Xiaochaihu Granule.