接續上一篇，我把克萊姆森大學的研究從這篇開始翻譯，其中有些圖是指向其他頁，為求閱讀方便也擷取到這一篇。
Clemson University 克萊姆森大學
克萊姆森大學的 John Lawson 博士及其研究小組已經完成了青邊貽貝在動物模型和細胞培養方面的一些關節炎研究，尤其對評估免疫調節功能特別深入研究。在使用實驗鼠的單獨研究中，青邊貽貝是顯示出有效地減少類風濕性關節炎的發作並使其逆轉。
在使用實驗鼠的研究中，確認健康狀況後隨機分組，對照組的動物注射第二型膠原蛋白(C-II) ，實驗組動物則餵食青邊貽貝(每天100mg / kg體重)。分析兩組中關節炎的發病率、關節炎的發病時間和嚴重程度，以及對第二型膠原蛋白的 IgG 和 IgM 抗體水平。餵食青邊貽貝的18隻動物中只有3隻(16.6％)發生關節炎，而在對照組中注射第二型膠原蛋白的15隻動物，有10隻(66.6％)發生嚴重的關節炎症（圖2）。
Research at Clemson University
John Lawson, PhD, and his research group at Clemson University have completed a number of arthritic studies with Perna in both animal models and in cell cultures, with a special interest in evaluating immune regulation.In separate studies using both rats and mice, Perna was shown to effectively reduce the onset of rheumatoid arthritis and to reverse it as well.
In a study using outbred rats, both control and test animals were injected with collagen II (C-II) to induce arthritis.The test animals were fed Perna (100mg/kg of body weight per day). Incidence of arthritis, the timing and severity of arthritis and IgG and IgM antibody levels to collagen II, in both groups, were analyzed. Only three of the eighteen test animals ( 16.6 % ) that were fed Perna developed arthritis, while in the control group, 10 out of 15 animals ( 66.6 % ) developed severe arthritic inflammation (Figure 2).
克萊姆森的結果與先前發表的報導一致，青邊貽貝可以產生抗發炎反應。與第二型膠原蛋白C-II 對照相比，使用青邊貽貝的實驗大鼠可延遲的關節炎發作。對於關節的測量（第二型膠原蛋白C-II 對照組關節炎關節的平均為19.5mm，而青邊貽貝組為14.2mm），顯示使用第二型膠原蛋白C-II 對照組動物的關節炎比較嚴重。青邊貽貝組和對照組之間的 IgG 和 IgM 抗體反應沒有顯著差異。但青邊貽貝組有效減少實驗鼠關節炎病變的發炎。
The Clemson results are consistent with previous published reports that Perna mussel produces an anti- inflammatory response. The rats which were on Perna prior to C-II induction of arthritis had a delayed onset of arthritis (one day later) as compared to arthritic controls. Based on joint measurement (average paw size for control arthritic joints was 19.5 mm as compared to 14.2 mm in the Perna group), the arthritis was also shown to be more severe in the control animals as compared to the animals in the Perna fed group that developed arthritis. There was no significant difference in the IgG and IgM antibody response between the Perna treated animals and the control group. Perna was also effective in decreasing inflammation of arthritic lesions in mice in which arthritis was induced by the C-II method.
該研究的下一階段中，把確診有關節炎症狀的實驗鼠，隨機分配到兩個治療組。第一組每天餵食青邊貽貝1.2g，而第二組不接受青邊貽貝補充劑來作為對照組。在107天的時間內對實驗鼠進行 0-3 的評分，作為關節炎症的測量。兩組均以 2.1 的相同平均得分開始研究。圖3）。將第107天的終點得分平均並顯示在表1中。
Table 1: End-point severity inflammation scores in arthritic mice after 107 days
Treatment
|
Score
|
Percent
|
Control
|
0
|
0%
|
Control
|
1
|
13.3%
|
Control
|
2
|
26.7%
|
Control
|
3
|
60%
|
Perna
|
0
|
7.1%
|
Perna
|
1
|
57.1%
|
Perna
|
2
|
28.6%
|
Perna
|
3
|
7.1%
|
Joint Inflammation Scores (0=No Inflammation, 3=Severe Inflammation)
對照組動物以平均得分2.65結束研究。該組中的大多數小鼠平均每日發炎評分在2.5至3.0之間。這些小鼠在所有四肢中都表現出嚴重的腫脹和水腫。相反，接受青邊貽貝的那些動物的平均得分為1.42，發炎症狀十分少，通常僅限於一個局部關節。令人驚訝的是在研究結束時，青邊貽貝組中的這些動物沒有表現出明顯的關節炎症狀。
In this study mice were randomly assigned to two treatment groups after the development of arthritic symptoms. The first group received standard mouse chow mixed with Perna daily, while the second group received no Perna supplement and therefore served as a control. The Perna fed mice were give 1.2 g Perna per day during the course of the study. Mice were scored in a range from 0-3 as a measurement of joint inflammation over a 107-day period. Both groups began the study with the same average score of 2.1 Figure 3). The endpoint scores at day 107 were averaged and presented in Table 1. The control animals ended the study with an average score of 2.65. The majority of the mice in this group were averaging a daily inflammation score of between 2.5 to 3.0. These mice exhibited severe swelling and edema in all four limbs. In contrast, those animals receiving Perna had an average score of 1.42. These animals displayed minimal inflammation, usually confined to only one limb. Surprisingly a few of the animals in the Perna-fed group displayed no apparent arthritic symptoms at the end of the study.
這次的研究除了確定青邊貽貝可有效預防關節炎以及逆轉病情，更進一步發現青邊貽貝可減少軟骨細胞凋亡、退化的狀況。第二型膠原蛋白C-II 對照組在實驗中軟骨細胞持續凋亡，比非關節炎的實驗鼠高約25.6％，但餵養青邊貽貝的實驗動物中，細胞凋亡率卻是顯著下降6.3％。在人體的初步研究中，我們已經顯示在骨關節炎和類風濕性關節炎中存在異常的軟骨細胞凋亡，類似於在關節炎小鼠中觀察到的。未來的研究將尋求確定青邊貽貝是否能讓患有關節炎的人降低細胞凋亡的水平。
The rat and mouse studies point out that Perna mussel may be effective in preventing arthritis as well as reversing the condition. Perna mussel also caused a decrease in apoptosis, or programmed cell death, in lymphocytes obtained from arthritic animals. Control arthritic mice exhibited an elevated background level of constant apoptosis of approximately 25.6 % above levels seen in nonarthritic controls. In Perna-fed animals, apoptosis fell significantly by an average of 6.3 % . In preliminary studies in humans, we have shown the presence of aberrant apoptosis in both osteoarthritis and rheumatoid arthritis similar to that observed in arthritic mice. Future studies will seek to determine if Perna modulates levels of apoptosis in humans suffering from arthritis.
接下來則是使用細胞培養方式的實驗，結果顯示青邊貽貝可作為免疫調節劑，調節身體及細胞免疫功能。
青邊貽貝提取物含有具備免疫調節活性的 Tween-20 extract。透過生物測定法來測量活性，將有青邊貽貝提取物處理和未處理的細胞清液，加入敏感的應答細胞中。這些應答細胞的增殖(增加)或抑制(減少)就會顯示對青邊貽貝提取物反應的程度。
用青邊貽貝提取物處理的細胞清液，加入具有腫瘤壞死因子(TNF-α)脂多醣複合物(LPS)的THP-1細胞24小時，再將清液加入L-929成纖維細胞中，觀察到腫瘤壞死因子TNF-α 生長受到抑制，青邊貽貝提取物降低腫瘤壞死因子TNF-α的水平，與未處理的對照相比約降低50％。
Cell culture results show that Perna also acts as an immunomodulating agent, affecting both humoral and cellular immunity.A Tween-20 extract of Perna with immunomodulating activity was prepared. The activity was measured by bioassays in which supernatants of Perna-treated and untreated immune cells were added to responder cells sensitive to an individual cytokine. Proliferation (increase) or inhibition (decrease) of these responder cells signaled the extent of cytokine production. THP-1 cells, which produced tumor necrosis factor- alpha (TNF-α) with lipopolysaccharide complex (LPS), were treated with Perna for 24 hours. The supernatants were added to L-929 fibroblast cell lines, whose growth is inhibited in the presence of TNF-α. Perna decreased the levels of TNF-α. produced by about 50 % compared to untreated controls.
類似情況發生在另一個細胞實驗，青邊貽貝降低了 LS174 模型中的炎症細胞因子(IL-1，IL-2，IL-6，腫瘤壞死因子TNF-α)。此外，青邊貽貝提取物抑制了幾種B類腫瘤細胞中抗體的產生。如下圖4所示，隨著青邊貽貝濃度的增加，發炎性細胞因子的濃度被抑制、降低。
Similarly Perna lowered the levels of interleukin-1 IL- 1) production in U-937 cell lines interleukin-2 (IL-2) in both Jurkat E6- 1 and EL-4 cell lines and interleukin-6 IL-6) in LS174T cell lines (Figures 4-9). In addition, the above extract of Perna inhibited the production of antibody in several B cell hybridoma cell lines. As shown by these figures, as the concentration of an inflammatory cytokine decreases concomitant with increases in added Perna concentrations, the viability of the responder cell lines either increase or decrease dependant on their specific response to the individual cytokine.
青邊貽貝提取物可以除去蛋白水解酶來調節免疫活性 (圖9：使用青邊貽貝提取物5mg，每兩小時觀察，連續12小時都有降低發炎狀況)。當然未來需要更多的研究，來確定這種下降是否與炎症細胞因子（IL-1，IL-2，IL-6，腫瘤壞死因子-α）的下調和異常免疫細胞（細胞凋亡）的減少有關，這些研究將有助於回答這個問題。
最近對系統性紅斑狼瘡（SLE）實驗鼠的一項研究顯示，青邊貽貝和N N-Dimethylglycine (DMG) 的組合，可有效地幫助逆轉該疾病的幾種臨床標誌物（參見Townsend Letter，2000年5月）。這些包括幾種發炎性細胞因子（包括IL-6和TNF-α）水平的顯著降低，以及單鍊和雙鏈DNA的抗核抗體。初步結果看起來非常令人鼓舞，青邊貽貝計劃在該領域開展更多研究。
All immunomodulatory activity of Perna mussel extract could be removed with proteolytic enzymes (Figure 9). More research is needed to determine whether this decrease corresponds to the down-regulation of inflammatory cytokines (IL-1, IL-2, IL- 6, Tumor Necrosis Factor-alpha) and a decrease of aberrant immune cells (apoptosis) that are contributing to the disease. A human study is now planned which will help to answer this question. A recent study in mice with systemic lupus erythematosis (SLE) has shown that a combination of Perna mussel and N N-Dimethylglycine (DMG) was effective in helping reverse several clinical markers for the disease (see Townsend Letter, May 2000). These include significant decreases in the levels of several inflammatory cytokines (including IL-6 and TNF-alpha) and in antinuclear antibodies to both single and double stranded DNA.Preliminary results look very encouraging and more studies in this area are planned.
註：青邊貽貝提取物
紐西蘭特有種青邊貽貝，提取出的成分中以 GAG 醣胺聚醣最為矚目，因為一般的葡萄糖胺（Glucosamine）經過食用後大約6~8週才能轉換成 GAG，成為關節可用的營養，但是青邊貽貝提取物穩定存在豐富的 GAG可直接被關節使用，當然青邊貽貝提取物中含有與人體相似的天然礦物質與其他成分也有助於關節炎的整體改善。
圖片來源 My Swiss Life
青邊貽貝在生長曲線的高峰期(大約18個月)收成，以急速冷凍乾燥的方式加工成細粉，這種方式不會破壞營養與活性，因此完整保存蛋白質、複合碳水化合物、脂質、天然礦物質、氨基酸和粘多醣（GAG 醣胺聚醣），為身體尤其是關節提供營養。符合潔淨海域、保存完整營養的青邊貽貝提取物，就會有「ORIGINAL PERNA EXTRAKT」的驗證標章。
