Effect Of Ginsenoside Rd Of Apoptosis Of Neutrophil After Tuberculosis Infection And Its Mechanism

“Tuberculosis(TB) is still one of the major health problems in the world; about a third of the global population infected with mycobacterium tuberculosis(MTB). The TB epidemic situation is very serious in China and the number of cases in China is the second one after India in the world. WHO includes China into one of 22 high TB burden countries in the world; now, there are about 500 million people are infected with MTB in China; each year, there are about 1.45 million new TB cases, including more than 65 cases of sputum smear-positive positive TB patients. The primary drug resistance rate of TB in China is as high as 18.6% and China is the world’s high incidence area; it is very difficult to treat these patients”[1]. TB has become an important issue in the health and economic development of China.In China, ginseng has been widely used as a traditional Chinese herbal medicine for thousands of years. Ginsenoside is the most effective component in ginseng. There are three categories of g ginsenoside, respectively, diol one, triol one and oleanolic acid; currently, the former two kinds are considered to be the main active ingredients of ginseng [2]and they include the common Rb1, Rb2 Rb3, Rc, Rd, Re, Rg1, Rg2, Rh1 and glycosidicPT. The studies in recent years have shown that ginsenoside Rd has anti-apoptotic effect, anti-inflammatory effect, and can promote the phagocytosis of microglia, facilitate learning functions and anti-oxidative stress.After the body receives inflammation stimulation, neutrophils is among the first batch of innate immune cells migrating from the blood to the microbial infection. Neutrophil, as a full-time phagocyte, has a series of microbiology that receptors can identify opsonization and the microorganism without oposonization and promote the phagocytosis. Most of these microorganisms are rapidly killed after they mix with neutrophils phagosome and lysosomes. In addition, neutrophils also respond to microbial pattern recognition receptors, secrete chemokines(such as IP-10, MCP-1and MIP-1a/b) and proinflammatory cytokines(such as IFN-g and TNF-a) to attract other immune cells. The previous studies have shown that ginsenoside Rd has anti-apoptosis and anti-inflammatory effect; the project researches the influence of ginsenoside Rd on the apoptosis of normal neutrophils and TB infection neutrophil apoptosis change and the possible mechanism, as well as the influence on the functions of neutrophils.Part1: Influence of Ginsenoside Rd on Neutrophil Apoptosis in the TB Infection Neutrophils ModelObjective: to study the influence of ginsenoside Rd on neutrophil apoptosis in healthy adults’ peripheral blood and TB infection model. Method: healthy adults’ peripheral blood and neutrophils are respectively extracted and are divided into normal neutrophils group, human type mycobacterium TB attenuated(H37Ra) infection neutrophils model group and human subcutaneous injection BCG infection neutrophils group. CCK 8 method, flow cytometry and immunocytochemistry staining are sued to examine neutrophil cell vitality and cell apoptosis of the three groups. Results: compared with the control group, the apoptosis rate of groups with treatment of neutrophils ginsenoside Rd(100 um) reduces significantly and the cell vitality increases significantly. Conclusions: ginsenoside Rd can inhibit the apoptosis of neutrophil in normal and TB infection model, improve cell survival rate and prolong cell survival time.Part 2: Protection Mechanism of Ginsenoside Rd on TB Infection NeutrophilsObjectives: To further study the protection mechanism of ginsenoside Rd on TB infection neutrophils on the basis that ginsenoside Rd can inhibit apoptosis of normal neutrophils and neutrophils in TB infection model and extend the service life of neutrophils. Method: healthy adults’ peripheral blood and neutrophils are respectively extracted and are divided into normal neutrophils group and TB infection neutrophils model group; PI3K/AKT, ERK1/2 inhibitor LY294002/PD98059 and ginsenoside Rd are added; flow cytometry and immunocytochemistry staining are sued to examine neutrophil cell vitality and cell apoptosis of the groups. Results: the cell apoptosis rate of the neutrophils treated with LY294002/PD98059 and ginsenoside Rd becomes than that of purely treated with ginsenoside Rd. Conclusions: ginsenoside Rd can inhibit apoptosis of normal neutrophil and neutrophil in TB infection model possibly by PI3K/AKT and ERK1/2.Part 3: Effects of Ginsenoside Rd on Neutrophil FunctionsObjective: to observe the effect of ginsenoside Rd on neutrophils BCG or H37 Ra phagocytosis. Methods: H37Ra/BCG is used to infect neutrophils; flow cytometry is applied to test cell phagocytosis rate; the modified acid-fast staining method is applied to measure cells phagocytic index. Results: the phagocytosis rate and index of neutrophils in the ginsenoside Rd group(100 um) are obviously higher than that of the control group. Conclusions: ginsenoside Rd can improve the phagocytosis of neutrophils.